RU2844751C1 - Agent for gene therapy based on csrp3 (rich in cysteine and glycine protein 3) - Google Patents

Abstract

FIELD: biotechnology; medicine.

SUBSTANCE: present invention relates to biotechnology and can be used in medicine. Invention discloses a genetic construct capable of increasing the expression of LIM muscular protein (MLP) containing a polynucleotide coding said protein, operably linked to a promoter specific for myocardial cells, for example, MHCK7 or HTNNT2.

EFFECT: invention is applicable in gene therapy of the diseases associated with CSRP3 protein deficiency caused by defects in the gene coding it, for example, in heart failure, various types of cardiomyopathy, and so forth.

70 cl, 5 dwg, 6 tbl, 5 ex

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Patent Application No. 63/061,727, filed August 5, 2020, the contents of which are incorporated herein by reference in their entirety.

STATEMENT REGARDING THE SEQUENCE LISTING

The sequence listing associated with this application is provided in text format in lieu of a paper copy and is incorporated by reference into the description herein. The text file containing the sequence listing has the following name: ROPA_020_01WO_ST25.txt. The text file is approximately 120 KB in size, was generated on August 3, 2021, and submitted electronically via EFS-Web.

LEVEL OF TECHNOLOGY

The cysteine-glycine-rich protein 3 ( CSRP3 ) gene encodes the muscle LIM protein (MLP). Genetic defects in CSRP3 are associated with autosomal dominant cardiomyopathy, both hypertrophic cardiomyopathy (HCM) and dilated cardiomyopathy (DCM), since autosomal dominant mutations in different domains of the protein are associated with different phenotypes. Loss-of-function mutations that decrease MLP levels can cause protein mislocalization and proteasome-mediated degradation, resulting in disruption of normal signaling pathways in cardiac and skeletal muscle. Alterations in MLP levels or subcellular localization have also been associated with skeletal myopathies, including facioscapulohumeral muscular dystrophy, nemaline myopathy, and limb-girdle muscular dystrophy type 2B. Changes in the levels of the MLP-b protein isoform or dysregulation of the MLP:MLP-b ratio have been identified in patients with limb-girdle muscular dystrophy type 2A, Duchenne muscular dystrophy, and dermatomyositis.

Patients with CSRP3 present with a variety of symptoms depending on the specific mutation, but common symptoms include obstructive HCM or DCM, ventricular hypertrophy (with interventricular septum in the range of 14–32 mm), ventricular tachycardia, exercise intolerance, and angina. Moderate NYHA scores of I–II are common. Sudden cardiac death has been reported, for example, in a family carrying the C58G mutation. In one study, the majority of C58G carriers who underwent muscle biopsy complained of myalgias and cramps on exertion on admission.

There is an unmet need for therapy for CSRP3- associated diseases or disorders. This paper presents a gene therapy approach to address this need.

ESSENCE OF THE INVENTION

The present invention generally relates to a means of gene therapy for a disease or disorder, such as cardiac disease or disorder, using a vector expressing MLP or a functional variant thereof.

In one aspect, the present invention provides a polynucleotide comprising an expression cassette and optionally flanking inverted terminal repeats (ITRs) of an adeno-associated virus (AAV), wherein the polynucleotide comprises a polynucleotide sequence encoding a muscle LIM protein (MLP) or a functional variant thereof, operably linked to a promoter.

In some embodiments, the promoter is a heart-specific promoter.

In some embodiments, the promoter is a muscle-specific promoter.

In some embodiments, the promoter is a cardiomyocyte-specific promoter.

In some embodiments, the promoter is the MHCK7 promoter.

In some embodiments, the MHCK7 promoter has at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 31.

In some embodiments, the promoter is the cardiac troponin T (hTNNT2) promoter.

In some embodiments, the hTNNT2 promoter has at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 32.

In some embodiments, the expression cassette comprises exon 1 of the cardiac troponin T (hTNNT2) gene, wherein optionally the hTNNT2 promoter and exon 1 together have at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 32.

In some embodiments, the promoter is a ubiquitous promoter, optionally a CMV promoter or a CAG promoter.

In some embodiments, the expression cassette comprises a polyA signal.

In some embodiments, the polyA signal is human growth hormone (hGH) polyA.

In some embodiments, the expression cassette comprises a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), optionally a WPRE(x).

In some embodiments, the muscle LIM protein (MLP) or a functional variant thereof is MLP.

In some embodiments, the MLP is a human MLP.

In some embodiments, the MLP is MLP isoform A.

In some embodiments, the MLP has at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 1.

In some embodiments, the MLP is MLP isoform B.

In some embodiments, the MLP has at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 2.

In some embodiments, the MLP has at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 3.

In some embodiments, the MLP has at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 4.

In some embodiments, the polynucleotide sequence encoding the MLP is a cysteine-glycine-rich protein 3 ( CSRP3 ) polynucleotide.

In some embodiments, the CSRP3 polynucleotide is a human CSRP3 polynucleotide.

In some embodiments, the polynucleotide sequence encoding the MLP has at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 5.

In some embodiments, the polynucleotide sequence encoding the MLP has at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 7.

In some embodiments, the polynucleotide comprises at least about 2.4 kb, no more than about 2.6 kb, or from about 2.4 kb to about 2.6 kb.

In some embodiments, the polynucleotide comprises at least about 3.0 kb, no more than about 3.3 kb, or from about 3.0 kb to about 3.3 kb.

In some embodiments, the polynucleotide comprises at least about 2.4 kb, at least about 2.6 kb, at least about 3.0 kb, at least about 3.3 kb, at least about 3.5 kb, at least about 3.7 kb, at least about 3.9 kb, at least about 4.1 kb, or at least about 4.3 kb.

In some embodiments, the polynucleotide comprises at least about 2.6 kb, at least about 3.0 kb, no more than about 3.3 kb, no more than about 3.5 kb, no more than about 3.7 kb, no more than about 3.9 kb, no more than about 4.1 kb, no more than about 4.3 kb, or no more than about 4.5 kb.

In some embodiments, the expression cassette has at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NOs: 8-11.

In some embodiments, the polynucleotide has at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NOs: 12-15.

In some embodiments, the expression cassette is flanked by 5′ and 3′ inverted terminal repeats (ITRs), optionally ITRs from AAV2, optionally ITRs that have at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NOs: 20-26.

In some embodiments, the polynucleotide is self-complementary.

In some embodiments, the polynucleotide comprises an expression cassette and a sequence that is reverse complementary to the expression cassette.

In some embodiments, the expression cassette and the reverse complementary sequence of the expression cassette are flanked by 5′ and 3′ inverted terminal repeats (ITRs), optionally ITRs from AAV2, optionally ITRs that have at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 23 or SEQ ID NO: 26.

In another aspect, the present invention provides a gene therapy vector comprising a polynucleotide of the present invention.

In some embodiments, the gene therapy vector is a recombinant adeno-associated virus (rAAV) vector.

In some embodiments, the rAAV-based vector is AAV9 or a functional variant thereof.

In some embodiments, the rAAV vector comprises a capsid protein that has 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 77.

In some embodiments, the rAAV-based vector is AAVrh10 or a functional variant thereof.

In some embodiments, the rAAV vector comprises a capsid protein that has 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 79.

In some embodiments, the rAAV-based vector is AAV6 or a functional variant thereof.

In some embodiments, the rAAV vector comprises a capsid protein that has 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 78.

In some embodiments, the rAAV-based vector is AAVrh74 or a functional variant thereof.

In some embodiments, the rAAV vector comprises a capsid protein that has 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 80.

In some embodiments, the rAAV-based vector is a self-complementary AAV-based vector.

In another aspect, the present invention provides a method of treating and/or preventing a disease or disorder in a subject in need thereof, comprising administering to the subject a vector of the present invention.

In some embodiments, the disease or disorder is a cardiac disorder.

In some embodiments, the disease or disorder is heart failure.

In some embodiments, the disease or disorder is hypertrophic cardiomyopathy.

In some embodiments, the disease or disorder is dilated cardiomyopathy.

In some embodiments, the subject is a mammal.

In some embodiments, the subject is a primate.

In some embodiments, the subject is a human.

In some embodiments, the subject has a mutation in the CSRP3 gene that results in an amino acid substitution selected from C58G, L44P, S54R, E55G, and/or K69R, relative to human CSRP3 encoding a human MLP having the sequence of SEQ ID NO: 1.

In some embodiments, the vector is administered via intravenous injection, intracardiac injection, intracardiac infusion, and/or cardiac catheterization.

In some embodiments, administration increases MLP expression by at least about 5%.

In some embodiments, administration increases MLP expression by at least about 30%.

In some embodiments, administration increases MLP expression by at least about 70%.

In some embodiments, administration increases MLP expression by about 5% to about 10%.

In some embodiments, administration increases MLP expression by about 30% to about 50%.

In some embodiments, administration increases MLP expression by between about 70% and about 100%.

In some embodiments, the method allows for the treatment and/or prevention of a disease or disorder.

In another aspect, the present invention provides a pharmaceutical composition comprising a vector of the present invention.

In another aspect, the present invention provides a kit comprising a vector or pharmaceutical composition of the present invention and optionally instructions for use.

In another aspect, the present invention provides the use of a composition of the present invention in the treatment of a disease or disorder, optionally in accordance with any of the methods disclosed herein.

In another aspect, the present invention provides a composition of the present invention for use in the treatment of a disease or disorder, optionally according to any of the methods disclosed herein.

In another aspect, the present invention provides a method of expressing a muscle LIM protein (MLP) or a functional variant thereof, comprising contacting a cell with a vector of the present invention.

In some embodiments, the cell is a cardiomyocyte.

In some embodiments, the cardiomyocyte is a human cardiomyocyte.

In some embodiments, the promoter is the MHCK7 promoter, and wherein the level of expression of MLP is at least 2-fold higher than the level of expression of MLP in a cell transduced with a vector having the hTNNT2 promoter.

In some embodiments, the promoter is the MHCK7 promoter, and wherein the level of expression of MLP is 2-10 times greater than the level of expression of MLP in a cell transduced with a vector having the hTNNT2 promoter.

Various other aspects and embodiments are disclosed in the following detailed description. The present invention is limited solely by the appended claims.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS

Fig. 1 shows a vector diagram of a non-limiting example of a vector genome. The complete polynucleotide sequence of the vector genome is shown under SEQ ID NO: 12. The header portion is an expression cassette (SEQ ID NO: 8).

Fig. 2 shows a vector diagram of a non-limiting example of a vector genome. The complete polynucleotide sequence of the vector genome is shown under SEQ ID NO: 13. The header portion is an expression cassette (SEQ ID NO: 9).

Fig. 3 shows a vector diagram of a non-limiting example of a vector genome. The complete polynucleotide sequence of the vector genome is shown under SEQ ID NO: 14. The header portion is an expression cassette (SEQ ID NO: 10).

Fig. 4 shows a vector diagram of a non-limiting example of a vector genome. The complete polynucleotide sequence of the vector genome is shown under SEQ ID NO: 15. The header portion is an expression cassette (SEQ ID NO: 11).

Fig. 5A shows the expression of CSRP3 in transduced CHO-Lec2.

Figure 5B shows CSRP3 expression in transduced cardiomyocytes (differentiated cell line AC16 - Sigma-Aldrich®, Cat. #SCC109). Cells were transduced with an MOI of 3E5 from each vector; after 6 days, cell lysates were collected and Western blotted using a polyclonal antibody to CSRP3 (Thermo-Fisher® PA5-29155 1:1000).

DETAILED DESCRIPTION OF THE PRESENT INVENTION

The present invention provides gene therapy vectors for CSPRP3 that deliver a polynucleotide encoding MLP, as well as a method of use and other compositions and methods. Treatment of a CSPRP3-related disorder is complicated by the autosomal dominant nature of most forms of CSPRP3-related disorders and factors indicating that the level of protein expression and the balance between MLP isoforms are critical for normal function in healthy subjects. Moreover, the success of cardiac gene therapy cannot be predicted. Cardiomyocytes are a particularly difficult cell type to use for gene therapy. The compositions and methods disclosed herein address this issue.

Definitions

Section headings are for organizational purposes only and should not be construed as limiting the described subject matter to particular aspects or embodiments.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Suitable methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in the practice of the present invention. All publications, patent applications, patents and other references mentioned herein are expressly incorporated by reference in their entirety. In the event of a dispute, the present description, including definitions, will control. Furthermore, the materials, methods and examples described herein are illustrative only and are not intended to be limiting.

All publications and patents mentioned herein are herein incorporated by reference in their entireties, as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In the event of a dispute, this application, including any definitions herein, will control. However, the citation of any literature, article, publication, patent, patent publication, and patent application cited herein does not constitute and should not be taken as an admission or suggestion in any way that it constitutes valid prior art or forms part of the general knowledge in any country in the world.

In this specification, any concentration range, percentage range, ratio range, or whole number range shall be understood to include the value of any whole number within the stated range and, where appropriate, fractions thereof (e.g., one tenth and one hundredth of a whole number), unless otherwise specified. The term “about,” when immediately preceding a number or figure, means that the number or figure is within the range of plus or minus 10%. As used herein, singular terms shall be understood to refer to “one or more” of the listed components, unless otherwise specified. Use of an alternative (e.g., "or") shall be understood to mean either one or both, or any combination of alternatives. The term "and/or" shall be understood to mean either one or both alternatives. As used herein, the terms "include" and "contain" are used synonymously.

As used herein, the terms "identity" and "identical" refer, with respect to a portion of a polypeptide or polynucleotide sequence, to the percentage of exactly matching residues in the results of an alignment of such a "query" sequence with a "subject" sequence, such as the results of an alignment generated by the BLAST algorithm. Identity is calculated, unless otherwise noted, over the entire length of the subject sequence. Thus, a query sequence "shares at least x% identity" to a subject sequence if, in an alignment of the query sequence with the subject sequence, at least x% (rounded down) of the residues in the subject sequence align as an exact match to the corresponding residue in the query sequence. If the subject sequence has variable positions (e.g., residues marked with X) are considered a match if they align with any residue in the query sequence.

As used herein, the term "AAV-based vector" or "rAAV-based vector" refers to a recombinant vector containing one or more polynucleotides (or transgenes) of interest that are flanked by AAV terminal repeat (ITR) sequences. Such AAV-based vectors can be replicated and packaged into infectious viral particles when present in a host cell that has been transfected with a plasmid encoding and expressing the rep and cap gene products. Alternatively, AAV-based vectors can be packaged into infectious particles using a host cell that has been stably engineered to express the rep and cap genes.

As used herein, the term "AAV virion" or "AAV viral particle" or "AAV vector particle" refers to a viral particle that consists of at least one AAV capsid protein and an encapsidated AAV-based polynucleotide vector. As used herein, if the particle comprises a heterologous polynucleotide (i.e., polynucleotide other than the wild-type AAV genome, such as a transgene to be delivered into a mammalian cell), it is commonly referred to as an "AAV vector particle" or simply an "AAV-based vector." Thus, the production of an AAV vector particle necessarily involves the production of an AAV-based vector, since such a vector is contained within the AAV vector particle.

As used herein, the term "promoter" refers to a polynucleotide sequence capable of stimulating the initiation of transcription of RNA from a polynucleotide in a eukaryotic cell.

As used herein, the term “vector genome” refers to a polynucleotide sequence packaged by a vector (e.g., rAAV virion) including flanking sequences (in AAV, inverted terminal repeats). The terms "expression cassette" and "polynucleotide cassette" refer to the portion of the vector genome between the flanking ITR sequences. "Expression cassette" implies that the vector genome contains at least one gene encoding a gene product operably linked to an expression control element (e.g., promoter).

As used herein, the term "patient in need" or "subject in need" refers to a patient or subject at risk of developing or suffering from a disease, disorder, or condition that is treatable or ameliorated by a recombinant gene therapy vector or gene editing system disclosed herein. A patient or subject in need may, for example, be a patient or subject diagnosed with a heart-related disorder. The subject may have a mutation in the CSRP3 gene or a deletion of all or part of the CSRP3 gene or regulatory sequences of the gene that causes aberrant expression of the MLP protein. The terms "subject" and "patient" are used interchangeably herein. A subject treated with the methods described herein may be an adult or a child. Subjects may vary in age.

As used herein, the term "variant" or "functional variant" refers interchangeably to a protein that has one or more amino acid substitutions, insertions, or deletions compared to a parent protein that retains one or more required activities of the parent protein.

As used herein, the term “genetic disorder” refers to a partial or complete loss of function or aberrant activity of a gene. For example, a subject may suffer from a genetic disorder of gene expression or functionCSRP3, which reduces the expression of or results in loss or aberrant function of the MLP protein in at least some cells (e.g., cells of the heart) of the subject.

As used herein, the term “treatment” refers to the alleviation of one or more symptoms of a disease or disorder. The term “prevention” refers to the delay or interruption of the onset of one or more symptoms of a disease or disorder or to the slowing of the progression of an associatedCSRP3diseases or disorders, such as hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM) or skeletal myopathy.

[100]

MLP protein or polynucleotide

The present invention provides compositions and methods of use relating to the muscle protein molecule LIM (MLP). Various mutations in CSRP3 are known to be associated with hypertrophic cardiomyopathy (HCM) or dilated cardiomyopathy (DCM). Both hereditary and de novo mutations have been identified. In some cases, a heterozygous missense mutation is sufficient to cause the disease.

The polypeptide sequence of MLP is as follows:

MPNWGGGAKCGACEKTVYHAEEIQCNGRSFHKTCFHCMACRKALDSTTVAAHESEIYCKVCYGRRYGPKGIGYGQGAGCLSTDTGEHLGLQFQQSPK PARSVTTSNPSKFTAKFGESEKCPRCGKSVYAAEKVMGGGKPWHKTCFRCAICGKSLESTNVTDKDGELYCKVCYAKNFGPTGIGFGGLTQQVEKKE

(SEQ ID NO: 1).

The second isoform of MLP has the following polypeptide sequence: MPNWGGGAKCGACEKTVYHAEEIQCNGRSFHKTCFHCSPQSRHAQLPPATLPNSLRSLESPRSALDVASQSMLLRRLWEVASLGTRPVSAVPSVGRVWSPQMSLTKMGNFIAKFAMPKILAPRVLGLEALHNKWKRKNEEVRRFSDFLRA

(SEQ ID NO: 2).

Another MLP isoform has the following polypeptide sequence:

MPNWGGGAKCGACEKTVYHAEEIQCNGRSFHKTCFHCLC

(SEQ ID NO: 3).

Another MLP isoform has the following polypeptide sequence:

MPNWGGGAKCGACEKTVYHAEEIQCNGRSFHKTCFHCTLAQDLFPLCHLWEESGVHKC

(SEQ ID NO: 4).

In some embodiments, the MLP protein comprises a polypeptide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 1-4.

In some embodiments, the present invention provides a recombinant adeno-associated virus (rAAV) virion comprising a capsid and a vector genome, wherein the vector genome comprises a polynucleotide sequence encoding an MLP or a functional variant thereof operably linked to a promoter. In some embodiments, the present invention provides a recombinant adeno-associated virus (rAAV) virion comprising a capsid and a vector genome, wherein the vector genome comprises a polynucleotide sequence encoding an MLP operably linked to a promoter. The polynucleotide encoding the MLP may comprise a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to

ATGCCAAACTGGGGCGGAGGCGCAAAATGTGGAGCCTGTGAAAAGACCGTCTACCATGCAGAAGAAATCCAGTGCAATGGAAGGAGTTTCCACAAGACGTGTTTCCACTGCATGGCCTGCAGGAAGGCTCTTGACAGCACGACAG TCGCGGCTCATGAGTCGGAGATCTACTGCAAGGTGTGCTATGGGCGCAGATTGGCCCCAAAGGGATCGGGTATGGACAAGGCGCTGGCTGTCTCAGCACAGACACGGGCGAGCATCTCGGCCTGCAGTTCCAACAGTCCCCAAAG CCGGCACGCTCAGTTACCACCAGCAACCCTTCCAAATTCACTGCGAAGTTTGGAGAGTCCGAGAAGTGCCCTCGATGTGGCAAGTCAGTCTATGCTGCTGAGAAGGTTATGGGAGGTGGCAAGCCTTGGCACAAGACCTGTTTCC GCTGTGCCATCTGTGGGAAGAGTCTGGAGTCCACAAAATGTCACTGACAAAGATGGGGAACTTTATTGCAAAGTTTGCTATGCCAAAAATTTTGGCCCCACGGGTATTGGGTTTGGAGGCCTTACACAACAAGTGGAAAAGAAAGAA

(SEQ ID NO: 5).

Optionally, the polynucleotide sequence encoding the vector genome may comprise a Kozak sequence, including but not limited to GCCACCATGG (SEQ ID NO: 6). The Kozak sequence may overlap with the polynucleotide sequence encoding the MLP protein or a functional variant thereof. For example, the vector genome may comprise a polynucleotide sequence (with the Kozak sequence underlined) that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to

gccaccATGC

(SEQ ID NO: 7).

In some embodiments, the Kozak sequence is an alternative Kozak sequence comprising or consisting of any of the following:

(gcc)gccRccAUGG (SEQ ID NO: 16);

(gcc)gccRccAUGC (SEQ ID NO: 17);

AGNNAUGN;

ANNAUGG;

ANNAUGC;

ACCAUGG;

ACCAUGC;

GACACCAUGG (SEQ ID NO: 18) and

GACACCAUGC (SEQ ID NO: 19).

In some embodiments, the vector genome does not contain a Kozak sequence.

Vector genome

The AAV virions of the present invention comprise a vector genome. The vector genome may comprise an expression cassette (or a polynucleotide cassette for gene editing purposes without the need for expression of the polynucleotide sequence). Any suitable inverted terminal repeats (ITRs) may be used. The ITRs may be of the same serotype as the capsid or of a different serotype (e.g., ITR from AAV2 can be used).

In some embodiments, the 5' ITR comprises a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT

(SEQ ID NO: 20).

In some embodiments, the 5' ITR comprises a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to

GCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTTGTAGTTAATGATTAACCCGCCATGCTACTTATCTACGTA

(SEQ ID NO: 21).

In some embodiments, the 5' ITR comprises a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to

CTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTTGTAGTTAATGATTAACCCGCCATGCTACTTATCTACGTA

(SEQ ID NO: 22).

In some embodiments, the 5' ITR comprises a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to

Ttggccactccctctctgcgcgctcgctcgctcactgaggccgcccgggcaaagcccgggcgtcgggcgacctttggtcgcccggcctcagtgagcgagcgagcgcgcagagagggagtggccaactccatcactaggggttcct

(SEQ ID NO: 23).

In some embodiments, the 3' ITR comprises a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to

AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

(SEQ ID NO: 24).

In some embodiments, the 3' ITR comprises a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to

TACGTAGATAAGTAGCATGGCGGGTTAATCATTAACTACAAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGC

(SEQ ID NO: 25).

In some embodiments, the 3' ITR comprises a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to

aggaacccctagtgatggagactccctctctgcgcgctcgctcgctcactgaggccgggcgaccaaaggtcgcccgacgcccgggctttgcccgggcggcctcagtgagcgagcgagcgcgcagagagggagt

(SEQ ID NO: 26).

In some embodiments, the vector genome comprises one or more filler sequences that, for example, at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical

GCGGCAATTCAGTCGATAACTATAACGGTCCTAAGGTAGCGATTTAAATACGCGCTCTCTTAAGGTAGCCCCGGGACGCGTCAATTGACTACAAACCGAGTATCTGCAGAGGGCCCTGCGTATG (SEQ ID NO: 27);

CTTCTGAGGCGGAAAGAACCAGATCCTCTCTTAAGGTAGCATCGAGATTTAAATTAGGGATAACAGGGTAATGGCGCGGGCCGC (SEQ ID NO: 28) or

GTTACCCAGGCTGGAGTGCAGTGGCACATTTCTGCTCACTGCAACCTCCTCCTCCCTGGGTTC (SEQ ID NO: 29).

Promoters

In some embodiments, a polynucleotide sequence encoding an MLP protein or a functional variant thereof is operably linked to a promoter.

The present invention contemplates the use of various promoters. Promoters useful in embodiments of the present invention include, but are not limited to, the cytomegalovirus (CMV) promoter, the phosphoglycerate kinase (PGK) gene promoter, or a promoter sequence consisting of the CMV enhancer and portions of the chicken beta-actin gene and rabbit beta-globin (CAG) gene promoter. In some cases, the promoter may be a synthetic promoter. Exemplary synthetic promoters are provided in Schlabach et al. PNAS USA . 107(6):2538–43 (2010). In some embodiments, the promoter comprises a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to

ACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACT TGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCT ATTACCATGGTCGAGGTGAGCCCCACGTTCTGCTTCACTCTCCCCATCTCCCCCCCCTCCCCACCCCCAATTTTGTATTTATTTATTTTTAATTTATTTTGTGCAGCGATGGGGGCGGGGGGGGGGGGGGCGCGCGCCAGGCG GGGCGGGGCGGGGCGAGGGGCGGGGCGGGGCGAGGCGGAGAGGTGCGGCGGCAGCCAATCAGAGCGGCGCGCTCCGAAAGTTTCCTTTTATGGCGAGGGCGGCGGCGGCGGCGGCCCTATAAAAAGCGAAGCGCGCGGCGGGCGG

(SEQ ID NO: 30).

In some embodiments, a polynucleotide sequence encoding an MLP protein or a functional variant thereof is operably linked to an inducible promoter. The inducible promoter may be configured to cause transcriptional expression or not cause transcriptional expression of the polynucleotide sequence in response to the addition or accumulation of an agent or in response to the removal, degradation or dilution of the agent. The agent may be a drug. The agent may be tetracycline or one of its derivatives, including but not limited to doxycycline. In some cases, the inducible promoter is a tet-on promoter, a tet-off promoter, a chemically regulated promoter, a physically regulated promoter (i.e., promoter that responds to the presence or absence of light or to low or high temperatures). Inducible promoters include those inducible by heavy metal ions (such as the murine mammary tumor virus (mMTV) promoter or various growth hormone gene promoters) and promoters from phage T7, which are active in the presence of T7 RNA polymerase. This list of inducible promoters is not exhaustive.

In some cases, the promoter is a tissue-specific promoter, such as a promoter capable of driving expression in a cardiac cell to a greater extent than in a cell other than a cardiac cell. In some embodiments, the tissue-specific promoter is selected from any of a variety of cardiac cell-specific promoters, including but not limited to desmin (Des), alpha-myosin heavy chain (α-MHC), myosin light chain 2 (MLC-2), cardiac troponin C (cTnC), cardiac troponin T (hTNNT2), muscle creatine kinase (CK), and combinations of these promoter/enhancer regions, such as MHCK7. In some cases, the promoter is a ubiquitous promoter. "Ubiquitous promoter" refers to a promoter that is not tissue-specific under experimental or clinical conditions. In some cases, the ubiquitous promoter is any of the CMV, CAG, UBC, PGK, EF1-alpha, GAPDH, SV40, HBV, chicken beta-actin, and human beta-actin promoters.

In some embodiments, the promoter sequence is selected from Table 3. In some embodiments, the promoter comprises a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 31-51.

Table 3

PROMOTER SUBSEQUENCE SEQ ID NO: MHCK7 ACCCTTCAGATTAAAAATAACTGAGGTAAGGGCCTGGGTAGGGGAGGTGGTGTGAGACGCTCCTGTCTCTCCTCTATCTGCCCATCGGCCCTTTGGGGAGGAGGAATGTGCCCAAGGACTAAAAAAAGGCCATGGAGCCAGAGGGGCGAGGCAACAGACCTTTCATGGGCAAACCTTGGGGCCCTGCTGTC TAGCATGCCCCACTACGGGTCTAGGCTGCCCATGTAAGGAGGCAAGGCCTGGGACACCCGAGATGCCTGGTTATAATTAACCCAGACATGTGGCTGCCCCCCCCCCCAACACCTGCTCTCTAAAAATAACCCTGTCCCTGGTGGATCCCCTGCATGCGAAGATCTTCGAACAAGGCTGTGGGGGACTG AGGGCAGGCTGTAACAGGCTTGGGGGCCAGGGCTTATACGTGCCTGGGACTCCCAAAGTATTACTGTTCCATGTTCCCGGCGAAGGGCCAGCTGTCCCCCGCCAGCTAGACTCAGCACTTAGTTTAGGAACCAGTGAGCAAGTCAGCCCTTGGGGCAGCCCATACAAGGCCATGGGGCTGGGCAAGCTGCACG CCTGGGTCCGGGGTGGGCACGGTGCCCGGGCAACGAGCTGAAAGCTCATCTGCTCTCAGGGGCCCCTCCCTGGGGACAGCCCCTCCTGGCTAGTCACACCCTGTAGGCTCCTCTATATAACCCAGGGGCACAGGGGCTGCCCTCATTCTACCACCACCTCCACAGCACAGACAGACACTCAGGAGCCAGCCAG 31 Human cardiac troponin T promoter (minus exon 1)
hTnnT2 / HTNNT2 CTCAGTCCATTAGGAGCCAGTAGCCTGGAAGATGTCTTTACCCCCAGCATCAGTTCAAGTGGAGCAGCACATAACTCTTGCCCTCTGCCTTCCAAGATTCTGGTGCTGAGACTTATGGAGTGTCT TGGAGGTTGCCTTCTGCCCCCCAACCCTGCTCCCAGCTGGCCCTCCCAGGCCTGGGTTGCTGGCCTCTGCTTTATCAGGATTCTCAAGAGGGACAGCTGGTTTATGTTGCATGACTGTTCCCTGCA TATCTGCTCTGGTTTTAAATAGCTTATCTGAGCAGCTGGAGGACCACATGGGCTTATATGGCGTGGGGTACATGTTCCTGTAGCCTTGTCCCTGGCACCTGCCAAAATAGCAGCCAACACCCCCC ACCCCCACCGCCATCCCCCTGCCCCACCCGTCCCCTGTCGCACATTCCTCCCTCCGCAGGGCTGGCTCACCAGGCCCCAGCCCACATGCCTGCTTAAAGCCCTCTCCATCCTCTGCCTCACCCAGT 33 Human cardiac troponin T promoter (with exon 1, underlined)
hTnnT2 / HTNNT2 CTCAGTCCATTAGGAGCCAGTAGCCTGGAAGATGTCTTTACCCCCAGCATCAGTTCAAGTGGAGCAGCACATAACTCTTGCCCTCTGCCTTCCAAGATTCTGGTGCTGAGACTTATGGAGTGTCT TGGAGGTTGCCTTCTGCCCCCCAACCCTGCTCCCAGCTGGCCCTCCCAGGCCTGGGTTGCTGGCCTCTGCTTTATCAGGATTCTCAAGAGGGACAGCTGGTTTATGTTGCATGACTGTTCCCTGCA TATCTGCTCTGGTTTTAAATAGCTTATCTGAGCAGCTGGAGGACCACATGGGCTTATATGGCGTGGGGTACATGTTCCTGTAGCCTTGTCCCTGGCACCTGCCAAAATAGCAGCCAACACCCCCC ACCCCCACCGCCATCCCCCTGCCCCACCCGTCCCCTGTCGCACATTCCTCCCTCCGCAGGGCTGGCTCACCAGGCCCCAGCCCACATGCCTGCTTAAAGCCCTCTCCATCCTCTGCCTCACCCAGT CCCCGCTGAGACTGAGCAGACGCCTCCAGGATCTGTCGGCAG 32 Mouse α-cardiac myosin heavy chain (αMHC) promoter GGTACCGGATCCTGCAAGGTCACACAAGGGTCTCCACCCACCAGGTGCCCTAGTCTCAATTTCAGTTTCCATGCCTTGTTCTCACAATGCTGGCCTCCCCAGAGCTAATTTGGACTTTGTTTTTATTTCAAAAGGGCCTGAATGAGGAGTAGATCTTGTGCTACCCAGCT CTAAGGGTGCCCGTGAAGCCCTCAGACCTGGAGCCTTTGCAACAGCCCTTTAGGTGGAAGCAGAATAAAGCAATTTTCCTTAAAGCCAAAATCCTGCCTCTAGACTCTTCTTCTCTGACCTCGGTCCCTGGGCTCTAGGGTGGGGAGGTGGGGCTTGGAAGAAGAAGGTGG GGAAGTGGCAAAAGCCGATCCCTAGGGCCCTGTGAAGTTCGGAGCCTTCCCTGTACAGCACTGGCTCATAGATCCTCCTCCAGCCAAACATAGCAAGAAGTGATACCTCCTTTGTGACTTCCCCAGGCCCAGTACCTGTCAGGTTGAAACAGGATTTAGAGAAGCCTCTGA ACTCACCTGAACTCTGAAGCTCATCCACCAAGCAAGCACCTAGGTGCCACTGCTAGTTAGTATCCTACGCTGATAATATGCAGAGCTGGGCCACAGAAGTCCTGGGGTGTAGGAACTGACCAGTGACTTTTCAGTCGGCAAAGGTATGACCCCCTCAGCAGATGTAGTAAT GTCCCCTTAGATCCCATCCCAGGCAGGTCTCTAAGAGGACATGGGATGAGAGATGTAGTCATGTGGCATTCCAAACACAGCTATCCACAGTGTCCCTTGCCCCTTCCACTTAGCCAGGAGGACAGTAACCTTAGCCTATCTTTCTTCCTCCCCATCCTCCCAGGACACAC CCCCTGGTCTGCAGTATTCATTTCTTCCTTCACGTCCCCTCTGTGACTTCCATTTGCAAGGCTTTTGACCTCTGCAGCTGCTGGAAGATAGAGTTTGGCCCTAGGTGTGGCAAGCCATCTCAAGAGAAAGCAGACAACAGGGGGACCAGATTTTGGAAGGATCAGGAACTA AATCACTGGCGGGCCTGGGGGTAGAAAAAAGAGTGAGTGAGTCCGCTCCAGCTAAGCCAAGCTAGTCCCCGAGATACTCTGCCACAGCTGGGCTGCTCGGGGTAGCTTTAGGAATGTGGGTCTGAAAGACAATGGGATTGGAAGACATCTCTTTGAGTCTCCCCTCAACCC CACCTACAGACACACTCGTGTGTGGCCAGACTCCTGTTCAACAGCCCTCTGTGTTCTGACCACTGAGCTAGGCAACCAGAGCATGGGCCCTGTGCTGAGGATGAAGAGTTGGTTACCAATAGCAAAACAGCAGGGGAGGGAGAACAGAGAACGAAATAAGGAAGGAAGAA GGAAAGGCCAGTCAATCAGATGCAGTCAGAAGAGATGGGAAGCCAACACACAGCTTGAGCAGAGGAAACAGAAAAGGGAGAGATTCTGGGCATAAGGAGGCCACAGAAAGAAGAGCCCAGGCCCCCCAAGTCTCCTCTTTATACCCTCATCCCGTCTCCCAATTAAGCCC ACTCTTCTTCCTAGATCAGACCTGAGCTGCAGCGAAGAGACCCGTAGGGAGGATCACACTGGATGAAGGAGATGTGTGGAGAAGTCCAGGGAACCTAAGAGCCAGAGCCTAAAAGAGCAAGAGATAAAGGTGCTTCAAAGGTGGCCAGGCTGTGCACACAGAGGGTCGAGG ACTGGTGGTAGAGCCTCAAGATAAGGATGATGCTCAGAATGGGCGGGGGGGGGGATTCTGGGGGGGGGAGAGAGAAGGTGAGAAGGAGCCTGGAACAGAGAATCTGGAAGCGCTGGAAACGATACCATAAAGGGAAGAACCCAGGCTACCTTTAGATGTAAATCATGAAAG ACAGGGAGAAGGGAAGCTGGAGAGAGTAGAAGGACCCCGGGGCAAGACATTGAAGCAAGGACAAGCCAGGTTGAGCGCTCCGTGAAATCAGCCTGCTGAAGGCAGAGCCCTGGTATGAGCACCAGAACAGCAGAGGCTAGGGTTAATGTCGAGACAGGGAACAGAAGGTAG ACACAGGAACAGACAGACGGGGGAGCCAGGTAACAAAGGAATGGTCCTTCTCACCTGTGGCCAGAGCGTCCATCTGTGTCCACATACTCTAGAATGTTCATCAGACTGCAGGGCTGGCTTGGGAGGCAGCTGGAAAGAGTATGTGAGAGCCAGGGGAGACAAGGGGGGC CTAGGAAAGGAAGAAGAGGGCAAACCAGGCCACACAAGAGGGCAGAGCCCAGAACTGAGTTAACTCCTTCCTTGTTGCATCTTCCATAGGAGGCAGTGGGAACTCTGTGACCACCATCCCCCATGAGCCCCCACTACCCATACCAAGTTTGGCCTGAGTGGCATTCTAGGT TCCCTGAGGACAGAGCCTGGCCTTTGTCTCTTGGACCTGACCCAAGCTGACCCAATGTTCTCAGTACCTTATCATGCCCTCAAGAGCTTGAGAACCAGGCAGTGACATATTAGGCCATGGGCTAACCCTGGAGCTTGCACACAGGAGCCTCAAGTGACCTCCAGGGACACA GCTGCAGACAGGTGGCCTTTATCCCCAAAGAGCAACCATTTGGCATAGGTGGCTGCAAATGGGAATGCAAGGTTGAATCAGGTCCCTTCAAGAATACTGCATGCAAGACCTAAGACCCCTGGAGAGAGGGGTATGCTCCTGCCCCCACCCACCATAAGGGGAGTGAACTAT CCTAGGGGGCTGGCGACCTTGGGGAGACACCACATTACTGAGAGTGCTGAGCCCAGAAAAACTGACCGCCCTGTGTCCTGCCCACCTCCACACTCTAGAGCTATATTGAGAGGTGACAGTAGATAGGGTGGGAGCTGGTAGCAGGGAGAGTGTTCCTGGGTGTGAGGGTG TAGGGGAAAGCCAGAGCAGGGGAGTCTGGCTTTGTCTCCTGAACACAATGTCTACTTAGTTATAACAGGCATGACCTGCTAAAGACCCAACATCTACGACCTCTGAAAAGACAGCAGCCCTGGAGGACAGGGGTTGTCTCTGAGCCTTGGGTGCTTGATGGTGCCACAAAG GAGGGCATGAGTGTGAGTATAAGGCCCCAGGAGCGTTAGAGAAGGGCACTTGGGAAGGGGTCAGTCTGCAGAGCCCCTATCCATGGAATCTGGAGCCTGGGGCCAACTGGTGTAAATCTCTGGGCCTGCCAGGCATTCAAAGCAGCACCTGCATCCTCTGGCAGCCTGGGG AGGCGGAAGGGAGGCAACCCCCCACTTATACCCTTTCTCCCTCAGCCCCAGGATTAACACCTCTGGCCTTCCCCCTTCCCACCTCCCATCAGGAGTGGAGGGTTGCAGAGGGAGGGTAAAAACCTACATGTCCAAACATCATGGTGCACGATATATGGATCAGTATGTGTAG AGGCAAGAAAGGAAATCTGCAGGCTTAACTGGGTTAATGTGTAAAGTCTGTGTGCATGTGTGTGTGTCTGACTGAAAACGGGCATGGCTGTGCAGCTGTTCAGTTCTGTGCGTGAGGTTACCAGACTGCAGGTTTGTGTGTAAATTGCCCAAGGCAAAGTGGGTGAATCC CTTCCATGGTTTAAAGAGATTGGATGATGGCCTGCATCTCAAGGACCATGGAAAATAGAATGGACACTCTATATGTGTCTCTAAGCTAAGGTAGCAAGGTCTTTGGAGGACACCTGTCTAGAGATGTGGGCAACAGAGACTACAGACAGTATCTGTACAGAGTAAGGAGAG AGAGGAGGGGGTGTAGAATTCTCTTACTATCAAAGGGAAACTGAGTCGTGCACCTGCAAAGTGGATGCTCTCCCTAGACATCATGACTTTGTCTCTGGGGAGCCAGCACTGTGGAACTTCAGGTCTGAGAGAGTAGGAGGCTCCCCTCAGCCTGAAGCTATGCAGATAGCC AGGGTTGAAAGGGGGAAGGGAGAGCCTGGGATGGGAGCTTGTGTTGGAGGCAGGGGACAGATATTAAGCCTGGAAGAGAAGGTGACCCTTACCCAGTTGTTCAACTCACCCTTCAGATTAAAAATAACTGAGGTAAGGGCCTGGGTAGGGGAGGTGGTGTGAGACGCTC CTGTCTCTCCTCTATCTGCCCATCGGCCCTTTGGGGAGGAGGAATGTGCCCAAGGACTAAAAAAAGGCCATGGAGCCAGAGGGGCGAGGGCAACAGACCTTTCATGGGCAAACCTTGGGGCCCTGCTGTCCTCCTGTCACCTCCAGAGCCAAGGGATCAAAGGAGGAGGA GCCAGGACAGGAGGGAAGTGGGAGGGAGGGTCCCAGCAGAGGACTCCAAATTTAGGCAGCAGGCATATGGGATGGGATATAAAGGGGCTGGAGCACTGAGAGCTGTCAGAGATTTCTCCAACCCAGGTAAGAGGGAGTTTCGGGTGGGGGCTCTTCACCCACACCAGACCT CTCCCCACCTAGAAGGAAACTGCCTTTCCTGGAAGTGGGGTTCAGGCCGGTCAGAGATCTGACAGGGTGGCCTTCCACCAGCCTGGGAAGTTCTCAGTGGCAGGAGGTTTCCACAAGAAACACTGGATGCCCCTTCCCTTACGCTGTCTTCTCCATCTTCCTCCTGGGGAT GCTCCTCCCCGTCTTGGTTTATCTTGGCTCTTCGTCTTCAGCAAGATTTGCCCTGTGCTGTCCACTCCATCTTTCTCTACTGTCTCCGTGCCTTGCCTTGCCTTCTTGCGTGTCCTTCCTTTCCACCCATTTCTCACTTCACCTTTTCTCCCCTTCTCATTTGTATTCATC CTTCCTTCCTTCCTTCCTTCCTTCCTTCCTTCCTTCCTTCCTTCCTTTCTCCCTTCCTTCCTTCCTTCCTTCCTTCCTTCCTTCCTTCCTTCCTGTGTCAGAGTGCTGAGAATCACACCTGGGTTCCCACCCTTATGTAAACAATCTTCCAGTGAGCCACAGCTTCAGT GCTGCTGGGTGCTCTCTTACCTTTCCTCACCCCCTGGCTTGTCCTGTTCCATCCTGGTCAGGATCTCTAGATTGGTCTCCCAGCCTCTGCTACTCCTCTTCCTGCCTGTTCCTCTCTCTGTCCAGCTGCGCCACTGTGGTGCCTCGTTCCAGCTGTGGTCCACATTCTTCAG GATTCTCTGAAAAGTTAACCAGGTGAGAATGTTTCCCCTGTAGACAGCAGATCACGATTCTCCCGGAAGTCAGGCTTCCAGCCCTCTCTTTCTCTGCCCAGCTGCCCGGCACTCTTAGCAAACCTCAGGCACCCTTACCCCACATAGACCTCTGACAGAGAAGCAGGCACT TTACATGGAGTCCTGGTGGGAGAGCCATAGGCTACGGTGTAAAAGAGGCAGGGAAGTGGTGGTGTAGGAAAGTCAGGACTTCACATAGAAGCCTAGCCCACACCAGAAATGACAGACAGATCCCTCCTATCTCCCCCATAAGAGTTTGAGTCGACCCGCGGCCCGAATTG 34 Chicken cardiac troponin T (cTnT) promoter GGGATAAAAGCAGTCTGGGCTTTCACATGACAGCATCTGGGGCTGCGGCAGAGGGTCGGGTCCGAAGCGCTGCCTTATCAGCGTCCCCAGCCCTGGGAGGTGACAGCTGGCTGGCTTGTGTCAGCCCCTCGGGCACTCACGTATCTCCGTCCGACGGGTTTAAAATAGCAAAACTCTGAGGCCACACAATAGCTTGGGCTTATATG GGCTCCTGTGGGGGAAGGGGGAGCACGGAGGGGGCCGGGGCCGCTGCTGCCAAAATAGCAGCTCACAAGTGTTGCATTCCTCTCTGGGCGCCGGGCACATTCCTGCTGGCTCTGCCCGCCCCGGGGTGGGCGCCGGGGGGACCTTAAAGCCTCTGCCCCCCAAGGAGCCCTTCCCAGACAGCCGCCGGCACCCACCGCTCCGTGGGA 35 Human creatine kinase M (hCKM) CTCTCAGCCCTGGAAGTCCTTGCTCACAGCCGAGGCGCCGAGAGCGCTTGCTCTGCCCAGATCTGCGCGAGTCTGGCGCCCGCGCTCTGAACGGCGTCGCTGCCCAGCCCCCTTCCCCGGGAGGTGGGAGCGGCCA CCCAGGGCCCCGTGGCTGCCCTTGTAAGGAGGCGAGGCCCGAGGACACCCGAGACGCCCGGTTATAATTAACCAGGACACGTGGCGAACCCCCCTCCAACACCTGCCCCCGAACCCCCCCATACCCAGCGCCTCGG GTCTCGGCCTTTGCGGCAGAGGAGACAGCAAAGCGCCCTCTAAAAATAACTCCTTTCCCGGCGACCGAGACCCTCCCTGTCCCCCGCACAGCGGAAATCTCCCAGTGGCACCGAGGGGGCGAGGGTTAAGTGGGGG GGAGGGTGACCACCGCCTCCCACCCTTGCCCTGAGTTTGAATCTCTCCAACTCAGCCAGCCTCAGTTTCCCCTCCACTCAGTCCCTAGGAGGAAGGGGCGCCCAAGCGCGGGTTTCTGGGGTTAGACTGCCCTCCAT TGCAATTGGTCCTTCTCCCGGCCTCTGCTTCCTCCAGCTCACAGGGTATCTGCTCCTCCTGGAGCCACACCTTGGTTCCCCGAGGTGCCGCTGGGACTCGGGTAGGGGTGAGGGCCCAGGGGGCACAGGGGGAGCC GAGGGCCACAGGAAGGGCTGGTGGCTGAAGGAGACTCAGGGGCCAGGGGACGGTGGCTTCTACGTGCTTGGGACGTTCCCAGCCACCGTCCCATGTTCCCGGCGGGGGGCCAGCTGTCCCCACCGCCAGCCCAACT CAGCACTTGGTCAGGGTATCAGCTTGGTGGGGGGGCGTGAGCCCAGCCCCTGGGGGCGGCTCAGCCCATACAAGGCCATGGGGCTGGGCGCAAAGCATGCCTGGGTTCAGGGTGGGTATGGTGCGGGAGCAGGGAGG TGAGAGGCTCAGCTGCCCTCCAGAACTCCTCCCTGGGGACAACCCCTCCCAGCCAATAGCACAGCCTAGGTCCCCCTATATAAGGCCACGGCTGCTGGCCCTTCCTTTGGGTCAGTGTCACCTCCAGGATACAGACA 36 Human beta-actin (HuBa) GCCCAGCACCCCAAGGCGGCCAACGCCAAAACTCTCCCTCCTCCTCTTCCTCAATCTCGCTCTCGCTCTTTTTTTTTTTCGCAAAAGGAGGGGAGAGGGGTAAAAAAATGCTGCACTGTGCGGCG AAGCCGGTGAGTGAGCGGCGCGGGGCCAATCAGCGTGCGCCGTTCCGAAAGTTGCCTTTTATGGCTCGAGCGGCCGCGGCGGCGCCCTATAAAACCCAGCGGCGCGACGCGCCACCACCGCCGAGTC 37 Chicken Beta Actin (CBA) GGTCGAGGTGAGCCCCACGTTCTGCTTCACTCTCCCCATCTCCCCCCCCTCCCCACCCCCAATTTTGTATTTATTTATTTTTTAATTATTTTGTGCAGCGATGGGGGCGGGGGGGGGGGGGGCGCGCGCCCAGGCGGGGCG GGGCGGGGCGAGGGGCGGGGCGGGGCGAGGCGGAGAGGTGCGGCGGCAGCCAATCAGAGCGGCGCGCTCCGAAAGTTTCCTTTTATGGCGAGGGCGGCGGCGGCGGCGGCCCTATAAAAAGCGAAGCGCGCGGCGGGCGGGA 38 Cytomegalovirus (CMV) TGGTGATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCG 39 Cytomegalovirus (CMV) (second version) TAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCC ATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTA TGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACC CCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTGGTTTAGTGAACCGT 40 Cytomegalovirus (CMV) (third version) CGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTG GAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGG ACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCAT TGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCT 41 CAG Promoter (First Version) ACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACT TGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCT ATTACCATGGTCGAGGTGAGCCCCACGTTCTGCTTCACTCTCCCCATCTCCCCCCCCTCCCCACCCCCAATTTTGTATTTATTTATTTTTAATTTATTTTGTGCAGCGATGGGGGCGGGGGGGGGGGGGGCGCGCGCCAGGCG GGGCGGGGCGGGGCGAGGGGCGGGGCGGGGCGAGGCGGAGAGGTGCGGCGGCAGCCAATCAGAGCGGCGCGCTCCGAAAGTTTCCTTTTATGGCGAGGGCGGCGGCGGCGGCGGCCCTATAAAAAGCGAAGCGCGCGGCGGGCGG 42 CAG promoter (second version) CGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACT GCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCA TCGCTATTACCATGTCGAGGTGAGCCCCACGTTCTGCTTCACTCTCCCCCATCTCCCCCCCCTCCCCACCCCCAATTTTGTATTTATTTATTTTTTAATTATTTTGTGCAGCGATGGGGGCGGGGGGGGGGGGGGCGCGCGCCAGG CGGGGCGGGGCGGGGCGAGGGGCGGGGCGGGGCGAGGCGGAGAGGTGCGGCGGCAGCCAATCAGAGCGGCGCGCTCCGAAAGTTTCCTTTTATGGCGAGGGCGGCGGCGGCGGCGGCCCTATAAAAAGCGAAGCGCGCGGCGGGCG 43 Human EF1-alpha (EF1-α) CAACCTTTGGAGGCTAAGCCAGCAATGGTAGAGGGAAGATTCTGCACGTCCCTTCCAGGCGGCCTCCCCGTCACCACCCCCCCCAACCCGCCCCGACCGGAGCTGAGAGTAATTCATACAAAAGGACTCGCCCCTGCCTTGGGGAATCCCAGGGACCGTCGTTAAACTCCCACTAACGTAGAACCCAGAGATCGCTGCGTTCCCGCCCCCTCACCCGCCCGCTCTCGT CATCACTGAGGTGGAGAATAGCATGCGTGAGGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGGAGGGGTCGGCAATTGAACGGGT GCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTT 44 Human Synapsin1 (Syn), short version AGTGCAAGTGGGTTTTAGGACCAGGATGAGGCGGGGTGGGGGTGCCTACCTGACGACCGACCCCGACCCACTGGACAAGCACCCAACCCCCATTCCCCAAATTGCGCATCCCCTATCAGAGAGGGGGGAGGGGAAACAGGATGCGGCGAGGCGCGTGCGCACTGCCAGCTTCAGCACCGCGGACAGTGCCTTCGCCCCCGC CTGGCGGCGCGCCACCGCCGCCTCAGCACTGAAGGCGCGCTGACGTCACTCGCCGGTCCCCCGCAAACTCCCCTTCCCGGCCACCTTGGTCGCGTCCGCGCCGCCGCCGGCCCAGCCGGACCGCACCACGCGAGGCGCGAGATAGGGGGGCACGGGCGCGACCATCTGCGCTGCGGCGCCGGCGACTCAGCGCTGCCTC 45 Human synapsin1 (Syn) with 3′ extension AGTGCAAGTGGGTTTTAGGACCAGGATGAGGCGGGGTGGGGGTGCCTACCTGACGACCGACCCCGACCCACTGGACAAGCACCCAACCCCCATTCCCCAAATTGCGCATCCCCTATCAGAGAGGGGGGAGGGGAAACAGGATGCGGCGAGGCGCGTGCGCACTGCCAGCTTCAGCACCGCGGACAGTGCCTTCGCCCCCGCCTGGCGGCGCGCGCCACCGCCGCC TCAGCACTGAAGGCGCGCTGACGTCACTCGCCGGTCCCCCGCAAACTCCCCTTCCCGGCCACCTTGGTCGCGTCCGCGCCGCCGCCGGCCCAGCCGGACCGCACCACGCGAGGCGCGAGATAGGGGGGCACGGGCGCGACCATCTGCGCTGCGGCGCCGGCGACTCAGCGCTGCCTCAGTCTGCGGTGGGCAGCGGAGGAGTCGTGTCGTGCCTGAGAGCGCAG 46 Human Synapsin1 (Syn) with 5' extension CTGCAGAGGGCCCTGCGTATGAGTGCAAGTGGGTTTTAGGACCAGGATGAGGCGGGGTGGGGGTGCCTACCTGACGACCGACCCCGACCCACTGGACAAGCACCCAACCCCCATTCCCCAAATTGCGCATCCCCTATCAGAGAGGGGGGAGGGGAAACAGGATGCGGCGAGGCGCGTGCGCACTGCCAGCTTCAGCACCGCGGACAGTGCCT TCGCCCCCGCCTGGCGGCGCGCGCCACCGCCGCCTCAGCACTGAAGGCGCGCTGACGTCACTCGCCGGTCCCCCGCAAACTCCCCTTCCCGGCCACCTTGGTCGCGTCCGCGCCGCCGCCGGCCCAGCCGGACCGCACCACGCGAGGCGCGAGATAGGGGGGCACGGGCGCGACCATCTGCGCTGCGGCGCCGGCGACTCAGCGCTGCCTC 47 Human CamKIIa (CaMKIIa) ACTTGTGGACAAAGTTTGCTCTATTCCACCTCCTCCAGGCCCTCCTTGGGTCCATCACCCCAGGGGTGCTGGGTCCATCCCACCCCCAGGCCCACACAGGCTTGCAGTATTGTGTGCGGTATGGTCAGGGCGTCCGAGAG CAGGTTTCGCAGTGGAAGGCAGGCAGGTGTTGGGGAGGCAGTTACCGGGGCAACGGGAACAGGGCGTTTTGGAGGTGGTTGCCATGGGGACCTGGATGCTGACGAAGGCTCGCGAGGCTGTGAGCAGCCACAGTGCCCTGC 48 eSYN promoter GACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGACTATTTA CGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGCTGCAGAGGGCCCTGCGTATGAGTGCAAGTGGGTTTTAGGACCA GGATGAGGCGGGGTGGGGGTGCCTACCTGACGACCGACCCCGACCCACTGGACAAGCACCCAACCCCCATTCCCCAAATTGCGCATCCCCTATCAGAGAGGGGGGAGGGGAAACAGGATGCGGCGAGGCGTCGCGACTGCCAGCTTCAGCACCGCGGACAGTGCCTTCGCCCCCGCCTGGCGGCGCGCGCCACCGCCGCCTCAGCACTGAAG GCGCGCTGACGTCACTCGCCGGTCCCCCGCAAACTCCCCTTCCCGGCCACCTTGGTCGCGTCCGCGCCGCCGCCGGCCCAGCCGGACCGCACCACGCGAGGCGCGAGATAGGGGGGCACGGGCGCGACCATCTGCGCTGCGGCGCCGGCGACTCAGCGCTGCCTCAGTCTGCGGTGGGCAGCGGAGGAGTCGTGTCGTGCCTGAGAGCGCAGG 49

In a preferred embodiment, the vector genome comprises a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 31. In a preferred embodiment, the vector genome comprises a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 32. In a preferred embodiment, the vector genome comprises a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 33.

Additional illustrative examples of promoters include the SV40 late promoter from simian virus 40, the baculovirus polyhedron enhancer/promoter element, the herpes simplex virus thymidine kinase gene (HSV tk) promoter, the cytomegalovirus (CMV) immediate early promoter, and various retroviral promoters, including LTR elements. A wide variety of other promoters are known and publicly available in the art, and the sequences of many such promoters are available in sequence databases such as the GenBank database.

Other regulatory elements

In some cases, the vectors of the present invention further comprise one or more regulatory elements selected from the group consisting of an enhancer, an intron, a poly-A signal, a sequence encoding peptide 2A, a WPRE (Woodchuck hepatitis virus post-transcriptional regulatory element) and an HPRE (hepatitis B post-transcriptional regulatory element).

In some embodiments, the vector comprises a CMV enhancer.

In certain embodiments, the vectors comprise one or more enhancers. In particular embodiments, the enhancer is a CMV enhancer sequence, a GAPDH enhancer sequence, a β-actin gene enhancer sequence, or an EF1-α enhancer sequence. The foregoing sequences are known in the art. For example, the sequence of the immediate early (IE) enhancer of CMV is as follows:

ACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGG CAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCA

(SEQ ID NO: 50).

In certain embodiments, the vectors comprise one or more introns. In certain embodiments, the intron is a rabbit globin intron sequence, a chicken β-actin intron sequence, a synthetic intron sequence, an SV40 intron sequence, or an EF1-α intron sequence.

In certain embodiments, the vectors comprise a polyA sequence. In certain embodiments, the polyA sequence is a rabbit globin polyA sequence, a human growth hormone polyA sequence, a bovine growth hormone polyA sequence, a PGK polyA sequence, an SV40 polyA sequence, or a TK polyA sequence. In some embodiments, the polyA signal may be a bovine growth hormone polyadenylation signal (bGHpA).

In certain embodiments, the vectors comprise one or more transcript stabilizing elements. In particular embodiments, the transcript stabilizing element is a WPRE sequence, an HPRE sequence, a nuclear backbone attachment region, a 3' UTR, or a 5' UTR. In particular embodiments, the vectors comprise both a 5' UTR and a 3' UTR.

In some embodiments, the vector comprises a 5' untranslated region (UTR) selected from Table 4. In some embodiments, the vector genome comprises a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 51-61.

Table 4

5 ' -NON-TRANSMITTABLE AREA SUBSEQUENCE SEQ ID NO: Human beta-actin exon/intron CGCGTCCGCCCGCGAGCACAGAGCCTCGCCTTTGCCGATCCGCCGCCCGTCCACACCCGCCGCCAGGTAAGCCCGGCCAGCCGACCGGGGCATGCGGCCGCGGCCCTTCGCCCGTGCAG AGCCGCCGTCTGGGCCGCAGCGGGGGGCGCATGGGGCGGAACCGGACCGCCGTGGGGGGCGCGGGAGAAGCCCCTGGGCCTCCGGAGATGGGGGACACCCCACGCCAGTTCGCAGGCGC GAGGCCGCGCTCGGGCGGGCGCGCTCCGGGGGTGCCGCTCTCGGGGCGGGGGCAACCGGCGGGGTCTTTGTCTGAGCCGGGCTCTTGCCAATGGGGATCGCACGGTGGGCGCGGCGTAG CCCCCGTCAGGCCCGGTGGGGGCTGGGGCGCCATGCGCTGTGCGCGCTGGTCCTTTGGGCGCTAACTGCGTGCGCGCTGGGAATTGGCGCTAATTGCGCGTGCGCGCTGGGACTCAATGG CGCTAATCGCGCGTGCGTTCTGGGGCCCGGGCGCTTGCGCCACTTCCTGCCCGAGCCGCTGGCGCCCGAGGGTGTGGCCGCTGCGTGCGCGCGCGCGACCCGGTCGCTGTTTGAACCGG GCGGAGGCGGGGCTGGCGCCCGGTTGGGAGGGGTTGGGGGCCTGGCTTCCTGCCGCGCGCCGCGGGGACGCCTCCGACCAGTGTTTGCCTTTTATGGTAATAACGCGGCCGGCCCGGCT TCCTTTGTCCCCAATCTGGGCGCGCGCCGGCGCCCCCTGGCGGCCTAAGGACTCGGCGCGCCGGAAGTGGCCAGGGCGGCAGCGGCTGCTCTTGGCGGCCCCGAGGTGACTATAGCCTT CTTTTGTGTCTTGATAGTTCGCCAGCCTCTGCTAACCATGTTCATGCCTTCTTCTTTTTCCTACAGCTCCTGGGCAACGTGCTGGTTATTGTGCTGTCTCATCATTTTGGCAAAGAATTC 51 Exon/intron of chicken beta actin + intron of rabbit globin GTCGCTGCGCGCTGCCTTCGCCCCGTGCCCCGCTCCGCCGCCGCCTCGCGCCGCCCGCCCCGGCTCTGACTGACCGCGTTACTCCCACAGGTGAGCGGGCGGGACGGCCCTTCTCCTCCGGGCTGTAATTAGC GCTTGGTTTAATGACGGCTTGTTTCTTTTCTGTGGCTGCGTGAAAGCCTTGAGGGGCTCCGGGAGGGCCCTTTGTGCGGGGGGAGCGGCTCGGGGGGTGCGTGCGTGTGTGTGTGCGTGGGGAGCGCCGCGTGC GGCTCCGCGCTGCCCGGCGGCTGTGAGCGCTGCGGGCGCGGCGCGGGGCTTTGTGCGCTCCGCAGTGTGCGCGAGGGGAGCGCGGCCGGGGGCGGTGCCCCGCGGTGCGGGGGGGGCTGCGAGGGGAACAAAG GCTGCGTGCGGGTGTGTGCGTGGGGGGGTGAGCAGGGGGTGTGGGCGCGTCGGTCGGGCTGCAACCCCCCCTGCACCCCCCTCCCCGAGTTGCTGAGCACGGCCCGGCTTCGGGTGCGGGGCTCCGTACGGGGG CGTGGCGCGGGGCTCGCCGTGCCGGGCGGGGGGTGGCGGCAGGTGGGGGTGCCGGGCGGGGCGGGGCCGCCTCGGGCCGGGGAGGGCTCGGGGGAGGGGCGCGGCGGCCCCCGGAGCGCCGGCGGCTGTCGAG GCGCGGCGAGCCGCAGCCATTGCCTTTTATGGTAATCGTGCGAGAGGGCGCAGGGACTTCCTTTGTCCCAAATCTGTGCGGAGCCGAAATCTGGGAGGCGCCGCCGCACCCCCTCTAGCGGGCGCGGGGCGAAG CGGTGCGGCGCCGGCAGGAAGGAAATGGGCGGGGAGGGCCTTCGTGCGTCGCCGCGCCGCCGTCCCCTTCTCCCTCTCCAGCCTCGGGGCTGTCCGCGGGGGGACGCTGCCTTCGGGGGGGACGGGGCAGGG CGGGGTTCGGCTTCTGGCGTGTGACCGGCGGCTCTAGAGCCTCTGCTAACCATGTTCATGCCTTCTTCTTTTTCCTACAGCTCCTGGGCAACGTGCTGGTTATTGTGCTGTCTCATCATTTTGGCAAAGAATTC 52 Chimeric intron sequence GGTAAGTTTAGTCTTTTTGTCTTTTATTTCAGGTCCCGGATCCGGTGGTGGTGCAAATCAAAGAACTGCTCCTCAGTGGATGTTGCCTTTACTTCTAGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCCGC 53 5'-UTR-Syn1 Hs AGTCTGCGGTGGGCAGCGGAGGAGTCGTGTCGTGCCTGAGAGCGCAGCTGTGCTCCTGGGCACCGCGCAGTCCGCCCCCGGCTCCTGGCCAGACCACCCCTAGGACCCCCTGCCCCAAGTCGCA 54 IE exon CMV TCAGATCGCCTGGAGAGGCCATCCACGCTGTTTTGACCTCCATAGTGGACACCGGGACCGATCCAGCCTCCGCGGCCGGGAACGGTGCATTGGAACGCGGATTCCCCGTGCCAAGAGTGAC 55 TPL-eMLP (adenoviral enhancer element) CTCACTCTCTTCCGCATCGCTGTCTGCGAGGGCCAGCTGTTGGGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCCAGTACTCTTGGATCGGAAACCCGTCGGCCTCCGAACGGTACTCCGCCAC CGAGGGACCTGAGCGAGTCCGCATCGACCGGATCGGAAAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCAAGGTAGGCTGAGCACCGTGGCGGGCGGCAGCGGGTGGCGGTCGGGGTTGTTT CTGGCGGAGGTGCTGCTGATGATGTAATTAAAGTAGGCGGTCTTGAGACGGCGGATGGTCGAGGTGAGGTGTGGCAGGCTTGAGATCCAGCTGTTGGGGTGAGTACTCCCTCTCAAAAGCGGGCATTA CTTCTGCGCTAAGATTGTCAGTTTCCAAAAACGAGGAGGATTTGATATTCACCTGGCCCGATCTGGCCATACACTTGAGTGACAATGACATCCACTTTGCCTTTCTCTCCACAGGTGTCCACTCCCAG 56 Human Ef1-α intron/exon CTTTTTCGCAACGGGTTTGCCGCCAGAACACAGGTAAGTGCCGTGTGTGGTTCCCGCGGGCCTGGCCTCTTTACGGGTTATGGCCCTTGCGTGCCTTGAATTACTTCCACCTGGCTCCA GTACGTGATTCTTGATCCCGAGCTGGAGCCAGGGGCGGGCCTTGCGCTTTAGGAGCCCCTTCGCCTCGTGCTTGAGTTGAGGCCTGGCCTGGGCGCTGGGGCCGCCGCGTGCGAATCTGG TGGCACCTTCGCGCCTGTCTCGCTGCTTTCGATAAGTCTCTAGCCATTTAAAATTTTTGATGACGTGCTGCGACGCTTTTTTTCTGGCAAGATAGTCTTGTAAATGCGGGCCAGGATCT GCACACTGGTATTTCGGTTTTTGGGCCCGCGGCCGGCGACGGGGCCCGTGCGTCCCAGCGCACATGTTCGGCGAGGCGGGGCCTGCGAGCGCGGCCACCGAGAATCGGACGGGGGTAGTC TCAAGCTGGCCGGCCTGCTCTGGTGCCTGGCCTCGCGCCGCCGTGTATCGCCCCGCCCTGGGCGGCAAGGCTGGCCCGGTCGGCACCAGTTGCGTGAGCGGAAAGATGGCCGCTTCCCG GCCCTGCTCCAGGGGGCTCAAAATGGAGGACGCGGCGCTCGGGAGAGCGGGCGGGTGAGTCACCCACACAAAGGAAAAGGGCCTTTCCGTCCTCAGCCGTCGCTTCATGTGACTCCACGG AGTACCGGGCGCCGTCCAGGCACCTCGATTAGTTCTGGAGCTTTTGGAGTACGTCGTCTTTAGGTTGGGGGGAGGGGTTTTATGCGATGGAGTTTCCCCACACTGAGTGGGTGGAGACT GAAGTTAGGCCAGCTTGGCACTTGATGTAATTCTCCTTGGAATTTGGCCTTTTTGAGTTTGGATCTTGGTTCATTCTCAAGCCTCAGACAGTGGTTCAAAGTTTTTTTCTTCCATTTCAG 57 Human EF1-α Intron A GTAAGTGCCGTGTGTGGTTCCCCGCGGGCCTGGCCTCTTTACGGGTTATGGCCCTTGCGTGCCTTGAATTACTTCCACCTGGCTGCAGTACGTGATTCTTGATCCCGAGCTTCGGGTT GGAAGTGGGTGGGAGAGTTCGAGGCCTTGCGCTTAAGGAGCCCCTTCGCCTCGTGCTTGAGTTGAGGCCTGGCCTGGGCGCTGGGGCCGCCGCGTGCGAATCTGGTGGCACCTTCGC GCCTGTCTCGCTGCTTTCGATAAGTCTCTAGCCATTTAAAATTTTTGATGACCTGCTGCGACGCTTTTTTTCTGGCAAGATAGTCTTGTAAATGCGGGCCAAGATCTGCACACTGGT ATTTCGGTTTTTGGGGCCGCGGGCGGCGACGGGGCCCGTGCGTCCCAGCGCACATGTTCGGCGAGGCGGGGCCTGCGAGCGCGCCACCGAGAATCGGACGGGGGTAGTCTCAAGCTG GCCGGCCTGCTCTGGTGCCTGGCCTCGCGCCGCCGTGTATCGCCCCGCCCTGGGCGGCAAGGCTGGCCCGGTCGGCACCAGTTGCGTGAGCGGAAAGATGGCCGCTTCCCGGCCCTGCTGCAGGGAGCTCAAAATGGAGGACGCGGCGCTCGGGAGAGCGGGCGGTGAGTCACCCACACAAAGGAAAAGGGCCTTTCCGTCCTCAGCCGTCGCTTCATGTGACTCCACGGAGTA CCGGGCGCCGTCCAGGCACCTCGATTAGTTCTCGAGCTTTTGGAGTACGTCGTCTTTAGGTTGGGGGGAGGGGTTTTATGCGATGGAGTTTCCCCACACTGAGTGGGTGGAGACTGA AGTTAGGCCAGCTTGGCACTTGATGTAATTCTCCTTGGAATTTGCCCTTTTTGAGTTTGGATCTTGGTTCATTCTCAAGCCTCAGACAGTGGTTCAAAGTTTTTTTCTTCCATTTCAG 58 5'-UTR of human CamKIIa TCAGAAGCCCCGGGCTCGTCAGTCAAACCGGTTCTCTGTTTGCACTCGGCAGCACGGGCAGGCAAGTGGTCCCTAGGTTCGGG 59 B-globin intron GTGAGTCTATGGGACCCTTGATGTTTTCTTTCCCCTTCTTTTCTATGGTTAAGTTCATGTCATAGGAAGGGGAGAAGTAACAGGGTACACATATTGACCAAATCAGGGTAATTTTGCAT TTGTAATTTTAAAAATGCTTTCTTCTTTTAATATACTTTTTTGTTTATCTTATTTCTAATACTTTCCCTAATCTCTTTCTTTCAGGGCAATAATGATACAATGTATCATGCCTCTTTG CACCATTCTAAAGAATAACAGTGATAATTTCTGGGTTAAGGCAATAGCAATATTTCTGCATATAAATATTTCTGCATATAAATTGTAACTGATGTAAGAGGTTTCATATTGCTAATAGCAGCTACAATCCAGCTACCATTCTGCTTTTATTTTATGGTTGGGATAAGGCTGGATTATTCTGAGTCCAAGCTAGGCCCTTTTGCTAATCATGTTCATACCTCTTATCTTCCTCCCACAG 60 Intron SV40 TCTAGAGGATCCGGTACTCGAGGAACTGAAAAACCAGAAAGTTAACTGGTAAGTTTAGTCTTTTTGTCTTTTATTTCAGGTCCCGGATCCGGTGGTGGTGCAAATCAAAGAACTGCTCCTCAGTGGATGTTGCCTTTACTTCTAGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCCGC 61

In some embodiments, the vector comprises a 3' untranslated region selected from Table 5. In some embodiments, the vector genome comprises a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 62-70.

Table 5

3 ' -NON-TRANSLATE AREA SUBSEQUENCE SEQ ID NO: WPRE(x) (woodchuck hepatitis mutant regulatory element - version 1) AATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTAT AAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCC CTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTGTGTTGCCACCTGGATT CTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGC 62 WPRE(x) (woodchuck hepatitis mutant regulatory element - version 2) TCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAA ATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCT CCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTGTGTTGCCACCTGGATTCT GCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCA 63 WPRE(x) (woodchuck hepatitis mutant regulatory element - version 3) TTCCTGTTAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTG TGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACT GACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCGCGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCCTCTTCGCCTTCGCCCTCAGACG AGTCGGATCTCCCTTTGGGCCGCCTCCCCGCCCATGTATCTTTTTCACCTGTGCCTTGTTTTTGCCTGTGTTCCGCGTCCTACTTTTCAAGCCTCCAAGCTGTGCCTTGGGCGGCTTTGGGGCATGGACATAGATCCCTATAAAGAATTTGGTTCATCTTATCAGTTGTTGAATTTTCTTCCTTGGAC 64 CAAX TGTGTGATAATG 65 EES CTGTTCTCATCACATCATATCAAGGTTATATACCATCAATATTGCCACAGATGTTACTTAGCCTTTTAATATTTCTCTAATTTAGTGTATATGCAATGATAGTTCTCTGATTTCTGAGATTGAGTTTCTCATGTGTAATGATTATTTAGAGTTTCTCTTTCATCTGTTCAAATTTTTGTCTAGTTTTATTTTTACTGATTT GTAAGACTTCTTTTTATAATCTGCATATTACAATTCTTTACTGGGGTGTTGCAAATATTTTCTGTCATTCTATGGCCTGACTTTTCTTAATGGTTTTTTTAATTTTAAAAATAAGTCTTAATATTCATGCAATCTAATTAACAATCTTTTCTTTGTGGTTAGGACTTTGAGTCATAAGAAATTTTTCTCTACACTGAAGTCA TGATGGCATGCTTCTATATTATTTTCTAAAAGATTTAAAGTTTTGCCTTCTCCATTTAGACTTATAATTCACTGGAATTTTTTTGTGTGTATGGTATGACATATGGGTTCCCTTTTATTTTTTACATATAAATATATTTCCCTGTTTTTCTAAAAAAGAAAAAGATCATCATTTTCCCATTGTAAAATGCCATATTTTTTTC ATAGGTCACTTACATATATCAATGGGTCTGTTTCTGAGCTCTACTCTATTTTATCAGCCTCACTGTCTATCCCCACACATCTCATGCTTTGCTCTAAATCTTGATATTTAGTGGAACATTCTTTCCCATTTTGTTCTACAAGAATATTTTTGTTATTGTCTTTGGGCTTTCTATATACATTTTGAAATGAGGTTGACAAGTTA 66 HPRE ATAACAGGCCTATTGATTGGAAAGTTTGTCAACGAATTGTGGGTCTTTTGGGGTTTGCTGCCCCTTTTACGCAATGTGGATATCCTGCTTTAATGCCTTTATATGCATGTATACAAGCAAAACAGGCTTTTACTTTCTCGCCAACTTACAAGGCCTTTCTCAGTAAACAGTATATGACCCT TTACCCCGTTGCTCGGCAACGGCCTGGTCTGTGCCAAGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCTTGGCCATAGGCCATCAGCGCATGCGTGGAACCTTTGTGTCTCCTCTGCCGATCCATACTGCGGAACTCCTAGCCGCTTGTTTTGCTCGCAGCAGGTCTGGAGCAAACCTCA TCGGGACCGACAATTCTGTCGTACTCTCCCGCAAGTATACATCGTTTCCATGGCTGCTAGGCTGTGCTGCCAACTGGATCCTGCGCGGGACGTCCTTTGTTTACGTCCCGTCGGCGCTGAATCCCGCGGACGACCCCTCCCGGGGCCGCTTGGGGCTCTACCGCCCGCTTCTCCGTCTGCC GTACCGTCCGACCACGGGGCGCACCTCTCTTTACGCGGACTCCCCGTCTGTGCCTTCTCATCTGCCGGACCGTGTGCACTTCGCTTCACCTCTGCACGTCGCATGGAGGCCACCGTGAACGCCCACCGGAACCTGCCCAAGGTCTTGCATAAGAGGACTCTTGGACTTTCAGCAATGTCATC 67 R2V17 (enhancer element derived from HepB) TTCCTGTAAACAGGCCTATTGATTGGAAAGTTTGTCAACGAATTGTGGGTCTTTTGGGGTTTGCTGCCCCTTTTACGCAATGTGGATATCCTGCTTTAATGCCTTTATATGCATGTATACAAGCAAAACAGGCTTTTACTTTCTCGCCAACTTACAAGGCCTTTCTCAGTAAACAGTATATGACCCTT TACCCCGTTGCTCGGCAACGGCCTGGTCTGTGCCAAGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCTTGGCCATAGGCCATCAGCGCATGCGTGGAACCTTTGTGTCTCCTCTGCCGATCCATACTGCGGAACTCCTAGCCGCTTGTTTTGCTCGCAGCTGGACTGGAGCAAACCCTCATCGGGACC GACAATTCTGTCGTACTCTCCCGCAAGCACTCACCGTTTCCGCGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCCTCTTCGCCTTCGCCCTCAGACG AGTCGGATCTCCCTTTGGGCCGCCTCCCCGCCCATGTATCTTTTTCACCTGTGCCTTGTTTTTGCCTGTGTTCCGCGTCCTACTTTTCAAGCCTCCAAGCTGTGCCTTGGGCGGCTTTGGGGCATGGACATAGATCCCTATAAAGAATTTGGTTCATCTTATCAGTTGTTGAATTTTCTTCCTTGGAC 68 3'-UTR (globin) GCTGGAGCCTCGGTAGCCGTTCCTCCTGCCCGCTGGGCCTCCCAACGGGCCCTCCTCCCCTCCTTGCACCGGCCCTTCCTGGTCTTTGAATAAA 69 WPRE(r) ATTCGAGCATCTTACCGCCATTTATTCCCATATTTGTTCTGTTTTTCTTGATTTGGGTATACATTTAAATGTTAATAAAACAAAATGGTGGGGCAATCATTTACATTTTTAGGGATATGTAATTACTAGTTCAGGTGTATTGCCACAAG ACAAACATGTTAAGAAACTTTCCCGTTATTTACGCTCTGTTCCTGTTAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGATATTCTTAACTATGTTGCTCCTTTTACGCTGTGTGGATATGCTGCTTTAATGCCTCTGTAT CATGCTATTGCTTCCCGTACGGCTTTCGTTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCCGTCAACGTGGCGTGGTGTGCTCTGTGTTTGCTGACGCAACCCCCACTGGCTGGGGCAT TGCCACCACCTGTCAACTCCTTTCTGGGACTTTCGCTTTCCCCCTCCCGATCGCCACGGCAGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTAGGTTGCTGGGCACTGATAATTCCGTGGTGTTGTCGGGGAAGGGCC 70

In some embodiments, the vector comprises a polyadenylation signal (polyA) selected from Table 6. In some embodiments, the polyA signal comprises a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 71-75.

Table 6

POLY-ADENYLATION SITE SUBSEQUENCE SEQ ID NO: Rabbit globin (pAGlobin-Oc) TGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGAACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATG AAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC 71 Bovine growth hormone (pAGH-Bt - version 1) TTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTA TTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAATACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGGGTACCCAGGTGCTGAAGAATTGACCCGGTTCCTCCTGGG 72 Bovine growth hormone (pAGH-Bt - version 2) TTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTA TTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGGGTACCCAGGTGCTGAAGAATTGACCCGGTTCCTCCTGGG 73 Bovine growth hormone (pAGH-Bt - version 3) CTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGG 74 Human growth hormone (pAGH-Hs) CTGCCCGGGTGGCATCCCTGTGACCCCTCCCCAGTGCCTCTCCTGGCCCTGGAAGTTGCCACTCCAGTGCCCACCAGCCTTGTCCTAATAAAATTAAGTTGCATCATTTTGTCTGACTAGGTGTCCTTCTATAATATTATGGGGTGGAGGGGGGTGGTATGGAGCAAGGGGCCCAAGTTGGGAAGAAACCTGTAGGGCCTGC 75

Exemplary vector genomes are depicted in Figs. 1-4 and are represented by SEQ ID NOs: 12-15. The expression cassette of each sequence, highlighted in capital letters, is represented by SEQ ID NOs: 8-11. In some embodiments, the vector genome comprises, consists essentially of, or consists of a polynucleotide sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NOs: 8-11, optionally with or without lowercase ITR sequences. The coding sequence is highlighted in capital letters.

Adeno-associated virus vector

Adeno-associated virus (AAV) is a replication-deficient parvovirus with a single-stranded DNA genome of approximately 4.7 kb in length, including two inverted terminal repeats (ITRs) of ~145 nucleotides. There are several known AAV variants, which are also sometimes referred to as serotypes when classified based on antigenic epitopes. The nucleotide sequences of the genomes of the AAV serotypes are known. For example, the complete genome of AAV-1 is submitted to GenBank under accession number NC_002077; the complete genome of AAV-2 is submitted to GenBank under accession number NC_001401 and Srivastava et al., J. Virol., 45: 555–564 (1983); the complete genome of AAV-3 is submitted to GenBank under accession number NC_1829; The complete genome of AAV-4 is submitted to GenBank under accession number NC_001829; the genome of AAV-5 is submitted to GenBank under accession number AF085716; the complete genome of AAV-6 is submitted to GenBank under accession number NC_00 1862; at least parts of the genomes of AAV-7 and AAV-8 are submitted to GenBank under accession numbers AX753246 and AX753249, respectively; the genome of AAV-9 is submitted to Gao et al., J. Virol., 78: 6381–6388 (2004); the genome of AAV-10 is submitted to Mol. Ther., 13(1): 67–76 (2006); and the genome of AAV-11 is submitted to Virology, 330(2): 375–383 (2004). The AAVrh.74 genome sequence is provided in U.S. Patent 9,434,928, incorporated herein by reference. The cis-acting sequences that direct viral DNA replication (rep), encapsulation/packaging, and integration of host cell chromosomes are contained within the AAV ITR. Three AAV promoters (named p5, p19, and p40 for their relative locations on the map) drive the expression of two internal AAV open reading frames encoding the rep and cap genes. The two rep promoters (p5 and p19), in combination with differential splicing of one AAV intron (at nucleotides 2107 and 2227), result in the production of four rep proteins (rep78, rep68, rep52, and rep40) from the rep gene. The rep proteins have multiple enzymatic properties that are ultimately responsible for replication of the viral genome. The cap gene is expressed under the control of the p40 promoter and encodes three capsid proteins: VP1, VP2, and VP3. Alternative splicing and nonconsensus translation start sites are responsible for the production of the three related capsid proteins. A single consensus polyadenylation site is located at position 95 of the AAV genome map. The life cycle and genetics of AAV are reviewed in Muzyczka, Current Topics in Microbiology and Immunology, 158: 97–129 (1992).

AAV has unique features that make it attractive as a vector for delivering foreign DNA into cells, such as for gene therapy. AAV infection of cells in culture is not cytopathic, and natural infection of humans and other animals is asymptomatic. Moreover, AAV infects a wide range of mammalian cells, allowing for targeting of a variety of tissues in vivo. Furthermore, AAV transduces slowly dividing and non-dividing cells and can persist for virtually the entire lifetime of these cells as a transcriptionally active nuclear episome (an extrachromosomal element). The AAV proviral genome has been inserted as cloned DNA into plasmids, allowing for the construction of recombinant genomes. Furthermore, because the signals directing AAV replication and genome encapsidation are contained within the ITR of the AAV genome, some or all of the internal ∼4.3 kb of AAV genome may be involved in the encapsidation of AAV. genome (encoding replication and structural capsid proteins, rep-cap) can be replaced by foreign DNA. To obtain AAV-based vectors, the rep and cap proteins can be presented in trans orientation. Another important feature of AAV is that it is an extremely stable and active virus. It easily withstands the conditions used to inactivate adenovirus (56° to 65°C for several hours), making cold storage of AAV less critical. AAV can even be lyophilized. Finally, AAV-infected cells are not resistant to superinfection.

Viral vectors for gene delivery useful in the practice of the present invention can be constructed using methodologies well known in the art of molecular biology. Typically, viral vectors carrying transgenes are assembled from polynucleotides encoding the transgene, suitable regulatory elements, and elements necessary for the production of viral proteins that mediate transduction of cells. Such recombinant viruses can be produced using techniques known in the art, for example, by transfection of packaging cells or transient transfection with helper plasmids or viruses. Typical examples of virus packaging cells include, but are not limited to, HeLa cells, SF9 cells (optionally with a baculovirus helper vector), 293 cells, etc. For the production of AAV-based vectors, the herpesvirus system as described in US20170218395A1 can be used. Detailed protocols for producing such replication-defective recombinant viruses can be found, for example, in WO95/14785, WO96/22378, U.S. Patent No. 5,882,877, U.S. Patent No. 6,013,516, U.S. Patent No. 4,861,719, U.S. Patent No. 5,278,056, and WO94/19478, the entire contents of each of which are incorporated herein by reference.

AAV vectors useful in practicing the present invention can be packaged into AAV virions (viral particles) using a variety of systems, including adenovirus and helper-less systems. Standard techniques in AAV biology include those described by Kwon and Schaffer. Pharm Res. (2008) 25(3):489–99; Wu et al. Mol. Ther. (2006) 14(3):316–27. Burger et al. Mol. Ther. (2004) 10(2):302–17; Grimm et al. Curr Gene Ther. (2003) 3(4):281–304; Deyle DR, Russell DW. Curr Opin Mol Ther. (2009) 11(4):442–447; McCarty et al. Gene Ther. (2001) 8(16):1248-54; and Duan et al. Mol Ther. (2001) 4(4):383-91. Helper-free systems included those described in US 6,004,797; US 7,588,772; and US 7,094,604;

The AAV DNA in the rAAV genomes may be from any AAV variant or serotype for which recombinant virus can be produced, including but not limited to the following AAV variants or serotypes: AAV-1, AAV-2, AAV-3, AAV-4, AAV-5, AAV-6, AAV-7, AAV-8, AAV-9, AAV-10, AAV-11, AAV-12, AAV-13, and AAVrh10. The production of pseudotyped rAAV is disclosed, for example, in WO 01/83692. Other types of rAAV variants are also contemplated, such as rAAV with mutations in the capsid. See, for example , Marsic et al., Molecular Therapy, 22(11): 1900-1909 (2014). The nucleotide sequences of the genomes of the various AAV serotypes are known in the art.

In some cases, the rAAV contains a self-complementary genome. As defined herein, an rAAV containing a “self-complementary” or “double-stranded” genome refers to an rAAV that has been engineered such that the coding region of the rAAV is configured to form an intramolecular double-stranded DNA template as described in McCarty et al. Self-complementary recombinant adeno-associated virus (scAAV) vectors promote efficient transduction independently of DNA synthesis. Gene Therapy. 8 (16): 1248-54 (2001). In some instances, the present invention contemplates the use of an rAAV containing a self-complementary genome because upon infection (such as transduction), rather than awaiting cell-mediated synthesis of the second strand of the rAAV genome, the two complementary halves of the scAAV will associate to form a single unit of double-stranded DNA (dsDNA) ready for immediate replication and transcription. It will be appreciated that, instead of the full coding capacity of an rAAV (4.7-6 kb), an rAAV containing a self-complementary genome may contain only approximately half that amount (≈2.4 kb).

In other instances, the rAAV-based vector comprises a single-stranded genome. As defined herein, a "single standard" genome refers to a genome that is not self-complementary. In most instances, non-recombinant AAVs have single-stranded DNA genomes. There has been some indication that rAAVs must be scAAVs to achieve efficient transduction of cells. However, the present invention contemplates rAAV-based vectors that may have single-stranded genomes rather than self-complementary genomes, with the understanding that other genetic modifications of the rAAV-based vector may be useful to achieve optimal gene transcription in target cells. In some instances, the present invention relates to single-stranded rAAV-based vectors capable of efficient gene transfer to the anterior segment of the mouse eye. See Wang et al. Single stranded adeno-associated virus achieves efficient gene transfer to anterior segment in the mouse eye. PLoS ONE 12(8): e0182473 (2017).

In some cases, the rAAV-based vector is of serotype AAV1, AAV2, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAV13, AAVrh10, or AAVrh74. The production of pseudotyped rAAV is disclosed, for example, in WO 01/83692. Other types of rAAV variants are also provided, such as rAAV with mutations in the capsid. See, for example , Marsic et al., Molecular Therapy, 22(11): 1900-1909 (2014). In some cases, the rAAV-based vector is of serotype AAV9. In some embodiments, said rAAV-based vector is of serotype AAV9 and comprises a single-stranded genome. In some embodiments, the rAAV-based vector is of the AAV9 serotype and comprises a self-complementary genome. In some embodiments, the rAAV-based vector comprises inverted terminal repeat (ITR) sequences from AAV2. In some embodiments, the rAAV-based vector comprises an AAV2 genome, such that the rAAV-based vector is an AAV-2/9-based vector, an AAV-2/6-based vector, or an AAV-2/8-based vector.

Full-length and capsid gene sequences for most of the known AAVs are provided in U.S. Patent No. 8,524,446, which is incorporated herein in its entirety.

AAV-based vectors may comprise a wild-type AAV sequence, or they may comprise one or more modifications to the wild-type AAV sequence. In certain embodiments, the AAV-based vector comprises one or more amino acid modifications, such as substitutions, deletions, or insertions, in a capsid protein, such as VP1, VP2, and/or VP3. In certain embodiments, the modification provides for reduced immunogenicity when the AAV-based vector is administered to a subject.

The rAAV capsid proteins can be modified such that the rAAV is targeted to a specific target tissue of interest, such as endothelial cells or, more particularly, endothelial tip cells. In some embodiments, the rAAV is administered directly into the intraventricular space of the subject.

In some embodiments, the rAAV virion is an AAV2 rAAV virion. The capsid may be an AAV2 capsid or a functional variant thereof. In some embodiments, the AAV2 capsid has at least 98%, 99%, or 100% identity to a reference AAV2 capsid, For example,

MAADGYLPDWLEDTLSEGIRQWWKLKPGPPPPKPAERHKDDSRGLVLPGYKYLGPFNGLDKGEPVNEADAAALEHDKAYDRQLDSGDNPYLKYNHADAEFQERLKEDTSFGGNLGRAVFQAKKRVLEPLGLVEEPVKTAPGKKRPVEHSPVEPDSSSGTGKAGQQPARKRLNFGQTGDADSVP DPQPLGQPPAAPSGLGTNTMATGSGAPMADNNEGADGVGNSSGNWHCDSTWMGDRVITTSTRTWALPTYNNHLYKQISSQSGASNDNHYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTQNDGTTTIANNLTSTVQVFTDSEYQLPYVLGSAHQGCLPPFPA DVFMVPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFTFSYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTNTPSGTTTQSR LQFSQAGASDIRDQSRNWLPGPCYRQQRVSKTSADNNNSEYSWTGATKYHLNGRDSLVNPGPAMASHKDDEEKFFPQSGVLIFGKQGSEKTN VDIEKVMITDEEEIRTTNPVATEQYGSVSTNLQRGNRQAATADVNTQGVLPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKHPP PQILIKNTPVPANPSTTFSAAKFASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYNKSVNVDFTVDTNGVYSEPRPIGTRYLTRNL

(SEQ ID NO: 76).

In some embodiments, the rAAV virion is an AAV9 rAAV virion. The capsid may be an AAV9 capsid or a functional variant thereof. In some embodiments, the AAV9 capsid has at least 98%, 99%, or 100% identity to a reference AAV9 capsid, For example,

MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPGNGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLK YNHADAEFQERLKEDTSFGGNLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAKKRLNFGQTGDTESVPD PQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFP ADVFMIPQYGYLTLNDGSQAVGRSSFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLSKTINGSGQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNNSEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDN VDADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPP PQILIKNTPVPADPPTAFNKDKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYSEPRPIGTRYLTRNL

(SEQ ID NO: 77).

In some embodiments, the rAAV virion is an AAV6 rAAV virion. The capsid may be an AAV9 capsid or a functional variant thereof. In some embodiments, the AAV6 capsid has at least 98%, 99%, or 100% identity to a reference AAV6 capsid, For example,

MAADGYLPDWLEDNLSEGIREWWDLKPGAPKPKANQQKQDDGRGLVLPGYKYLGPFNGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLR YNHADAEFQERLQEDTSFGGNLGRAVFQAKKRVLEPFGLVEEGAKTAPGKKRPVEQSPQEPDSSSGIGKTGQQPAKKRLNFGQTGDSESVPD PQPLGEPPATPAAVGPTTMASGGGAPMADNNEGADGVGNASGNWHCDSTWLGDRVITTSTRTWALPTYNNHLYKQISSASTGASNDNHYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTTNDGVTTIANNLTSTVQVFSDSEYQLPYVLGSAHQGCLPPFPA DVFMIPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFTFSYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLNRTQNQSGSAQNKD LLFSRGSPAGMSVQPKNWLPGPCYRQQRVSKTKTDNNNSNFTWTGASKYNLNGRESIINPGTAMASHKDDKDKFFPMSGVMIFGKESAGASN TALDNVMITDEEEIKATNPVATERFGTVAVNLQSSSTDPATGDVHVMGALPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKHPPPQILIKNTPVPANPPAEFSATKFASFITQYSTGQVSVEIEWELQKENSKRWNPEVQYTSNYAKSANVDFTVDNNGLYTEPRPIGTRYLTRPL

(SEQ ID NO: 78).

In some embodiments, the rAAV virion is an AAVrh.10 rAAV virion. The capsid may be an AAV9 capsid or a functional variant thereof. In some embodiments, the AAVrh.10 capsid has at least 98%, 99%, or 100% identity to a reference AAVrh.10 capsid, For example,

MAADGYLPDWLEDNLSEGIREWWDLKPGAPKPKANQQKQDDGRGLVLPGYKYLGPFNGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLR YNHADAEFQERLQEDTSFGGNLGRAVFQAKKRVLEPLGLVEEGAKTAPGKKRPVEPSPQRSPDSSTGIGKKGQQPAKKRLNFGQTGDSESVP DPQPIGEPPAGPSGLGSGTMAAGGGAPMADNNEGADGVGSSSGNWHCDSTWLGDRVITTSTRTWALPTYNNHLYKQISNGTSGGSTNDNTYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTQNEGTKTIANNLTSTIQVFTDSEYQLPYVLGSAHQGCLPPFP ADVFMIPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFEFSYQFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTQSTGGTAGTQ QLLFSQAGPNNMSAQAKNWLPGPCYRQQRVSTTLSQNNNSNFAWTGATKYHLNGRDSLVNPGVAMATHKDDEERFFPSSGVLMFGKQGAGKD NVDYSSVMLTSEEEIKTTNPVATEQYGVVADNLQQQNAAPIVGAVNSQGALPGMVWQNRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGLKHP PPQILIKNTPVPADPPTTFSQAKLASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSTNVDFAVNTDGTYSEPRPIGTRYLTRNL

(SEQ ID NO: 79).

In some embodiments, the capsid protein is encoded by a polynucleotide supplied on a plasmid in trans to a transfer plasmid. The polynucleotide sequence of the cap gene from wild-type AAVrh74 is as follows:

AAVrh74 capsid encoding sequence (SEQ ID NO: 80)

ATGGCTGCCGATGGTTATCTTCCAGATTGGCTCGAGGACAACCTCTCTGAGGGCATTCGCGAGTGGTGGGACCTGAAACCTGGAGCCCCGAAACCCAAAGCCAACCAGCAAAAGCAGGACAACGGCCGGGGTCTGGTG CTTCCTGGCTACAAGTACCTCGGACCCTTCAACGGACTCGACAAGGGGGAGCCCGTCAACGCGGCGGACGCAGCGGCCCTCGAGCACGACAAGGCCTACGACCAGCAGCTCCAAGCGGGTGACAATCCGTACCTGCGGT ATAATCACGCCGACGCCGAGTTTCAGGAGCGTCTGCAAGAAGATACGTCTTTTGGGGGCAACCTCGGGCGCGCAGTCTTCCAGGCCAAAAAGCGGGTTCTCGAACCTCTGGGCCTGGTTGAATCGCCGGTTAAGACGG CTCCTGGAAAGAAGAGACCGGTAGAGCCATCACCCCAGCGCTCTCCAGACTCCTCTACGGGCATCGGCAAGAAAGGCCAGCAGCCCGCAAAAAAGAGACTCAATTTTGGGCAGACTGGCGACTCAGAGTCAGTCCCCGA CCCTCAACCAATCGGAGAACCACCAGCAGGCCCCTCTGGTCTGGGATCTGGTACAATGGCTGCAGGCGGTGGCGCTCCAATGGCAGACAATAACGAAGGCGCCGACGGAGTGGGTAGTTCCTCAGGAAATTGGCATTG CGATTCCACATGGCTGGGCGACAGAGTCATCACCACCAGCACCCGCACCTGGGCCCTGCCCACCTACAACAACCACCTCTACAAGCAAATCTCCAACGGGACCTCGGGAGGAAGCACCAACGACAACACCTACTTCGGC TACAGCACCCCTGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTTTCACCACGTGACTGGCAGCGACTCATCAACAACAACTGGGGATTCCGGCCCAAGAGGCTCAACTTCAAGCTCTTCAACATCCAAGTCA AGGAGGTCACGCAGAATGAAGGCACCAAGACCATCGCCAATAACCTTACCAGCACGATTCAGGTCTTTACGACTCGGAATACCAGCTCCCGTACGTGCTCGGCTCGGCGCACCAGGGCTGCCTGCCTCCGTTCCCGGC GGACGTCTTCATGATTCCTCAGTACGGGTACCTGACTCTGAACAATGGCAGTCAGGCTGTGGGCCGGTCGTCCTTCTACTGCCTGGAGTACTTTCCTTCTCAAATGCTGAGAACGGGCAACAACTTTGAATTCAGCTA CAACTTCGAGGACGTGCCCTTCCACAGCAGCTACGCGCACAGCCAGAGCCTGGACCGGCTGATGAACCCTCTCATCGACCAGTACTTGTACTACCTGTCCCGGACTCAAAGCACGGGCGGTACTGCAGGAACTCAGCAG TTGCTATTTTCTCAGGCCGGGCCTAACAACATGTCGGCTCAGGCCAAGAACTGGCTACCCGGTCCCTGCTACCGGCAGCAACGCGTCTCCACGACACTGTCGCAGACAACAACAGCAACTTTGCCTGGACGGGTGCC ACCAAGTATCATCTGAATGGCAGAGACTCTCTGGTGAATCCTGGCGTTGCCATGGCTACCCACAAGGACGACGAAGAGCGATTTTTTCCATCCAGCGGAGTCTTAATGTTTGGGAAACAGGGAGCTGGAAAAGACAACG TGGACTATAGCAGCGTGATGCTAACCAGCGAGGAAGAAATAAAGACCACCAACCCAGTGGCCACAGAACAGTACGGCGTGGTGGCCGATAACCTGCAACAGCAAAACGCCGCTCCTATTGTAGGGGCCGTCAATAGTC AAGGAGCCTTACCTGGCATGGTGTGGCAGAACCGGGACGTGTACCTGCAGGGTCCCATCTGGGCCAAGATTCCTCATACGGACGGCAACTTTCATCCCTCGCCGCTGATGGGAGGCTTTGGACTGAAGCATCCGCCTCC TCAGATCCTGATTAAAAACACACCTGTTCCCGCGGATCCTCCGACCACCTTCAATCAGGCCAAGCTGGCTTCTTTCATCACGCAGTACAGTACCGGCCAGGTCAGCGTGGAGATCGAGTGGGAGCTGCAGAAGGAGAAC AGCAAACGCTGGAACCCAGAGATTCAGTACACTTCCAACTACTACAAATCTACAAATGTGGACTTTGCTGTCAATACTGAGGGTACTTATTCCGAGCCTCGCCCCATTGGCACCCGTTACCTCACCCGTAATCTGTAA.

The present invention further provides protein sequences for VP1, VP2 and VP3 of AAVrh74, including SEQ ID NOs: 2-4, and homologues or functional variants thereof.

VP1 AAVrh74 (SEQ ID NO: 81)

MAAGGGAPMADNNEGADGVGSSSGNWHCDSTWLGDRVITTSTRTWALPTYNNHLYKQISNGTSGGSTNDNTYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTQNEGTKTIAN NLTSTIQVFTDSEYQLPYVLGSAHQGCLPPFPADVFMIPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFEFSYNFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTQSTGGTAGTQQLLFSQAGPN NMSAQAKNWLPGPCYRQQRVSTTLSQNNNSNFAWTGATKYHLNGRDSLVNPGVAMATHKDDEERFFPSSGVLMFGKQGAGKDNVDYSSVMLTSEEEIKTTNPVATEQYGVVADNLQQQNAAPIVGAVNSQGALP GMVWQNRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGLKHPPPQILIKNTPVPADPPTTFNQAKLASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSTNVDFAVNTEGTYSEPRPIGTRYLTRNL.

VP2 AAVrh74 (SEQ ID NO: 82)

STIQVFTDSEYQLPYVLGSAHQGCLPPFPADVFMIPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFEFSYNFEDVPFHSSYAHSQSLDRLMNP LIDQYLYYLSRTQSTGGTAGTQQLLFSQAGPNNMSAQAKNWLPGPCYRQQRVSTTLSQNNNSNFAWTGATKYHLNGRDSLVNPGVAMATHKDDEERFFPS SGVLMFGKQGAGKDNVDYSSVMLTSEEEIKTTNPVATEQYGVVADNLQQQNAAPIVGAVNSQGALPGMVWQNRDVYLQGPIWAKIPHTDGNFHPSPLMGG FGLKHPPPQILIKNTPVPADPPTTFNQAKLASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSTNVDFAVNTEGTYSEPRPIGTRYLTRNL.

VP3 AAVrh74 (SEQ ID NO: 83)

RTGNNNFEFSYNFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTQSTGGTAGTQQLLFSQAGPNNMSAQAKNWLPGPCYRQQRVSTTLSQNNNSNFAWTGATKYHLNGRDSLVNPGVAMATHKDDEERFFPSSGVLMFGKQGAGKDNVDYSSVMLTSEEEIKTTN PVATEQYGVVADNLQQQNAAPIVGAVNSQGALPGMVWQNRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGLKHPPPQILIKNTPVPADPPTTFNQAKLASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSTNVDFAVNTEGTYSEPRPIGTRYLTRNL.

In certain cases, the AAVrh74 capsid comprises the amino acid sequence set forth in SEQ ID NO: 2. In some embodiments, the rAAV vector comprises a polypeptide that comprises, or consists essentially of, or further consists of a sequence that, for example, at least 65%, at least 70%, at least 75%, at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, or 89%, more typically 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more identical to the AAVrh74 VP1 amino acid sequence set forth in SEQ ID NO: 2. In some embodiments, the rAAV vector comprises a polypeptide that comprises, or consists essentially of, or further consists of, a sequence that, for example, at least 65%, at least 70%, at least 75%, at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, or 89%, more typically 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more identical to the AAVrh74 VP2 amino acid sequence set forth in SEQ ID NO: 3. In some embodiments, the rAAV vector comprises a polypeptide that comprises, or consists essentially of, or further consists of, a sequence that, for example, at least 65%, at least 70%, at least 75%, at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88% or 89%, more typically 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence of VP3 of AAVrh74, as set forth in SEQ ID NO: 4.

In some embodiments, the rAAV virion is an AAV-PHP.B rAAV virion or a neutrophic variant thereof, such as, but not limited to, one of the variants described in International Patent Publication Nos. WO 2015/038958 A1 and WO 2017/100671 A1. For example, the AAV capsid may comprise at least 4 contiguous amino acids from the sequence TLAVPFK (SEQ ID NO: 85) or KFPVALT (SEQ ID NO: 86), such as, inserted between the sequences encoding amino acids 588 and 589 in AAV9.

The capsid may be an AAV-PHP.B capsid or a functional variant thereof. In some embodiments, the AAV-PHP.B capsid has at least 98%, 99%, or 100% identity to a reference AAV-PHP.B capsid, For example,

MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPGNGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLK YNHADAEFQERLKEDTSFGGNLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAKKRLNFGQTGDTESVPDP QPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADV FMIPQYGYLTLNDGSQAVGRSSFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTINGSGQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNNSEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVDADK VMITNEEEIKTTNPVATESYGQVATNHQSAQTLAVPFKAQAQTGWVQNQGILPGMVWQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHP PPQILIKNTPVPADPPTAFNKDKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYSEPRPIGTRYLTRNL

(SEQ ID NO: 84).

Additional AAV capsids used in the rAAV virions of the present invention include those disclosed in Patent Publication Nos. WO 2009/012176 A2 and WO 2015/168666 A2.

Without being limited by any theory, the present inventors have determined that an AAV9-based vector, an AAVrh.74-based vector, or an AAVrh.10-based vector will provide the desired cardiac tropism to the vector. Without being limited by any theory, the present inventors have further determined that an AAV9-based vector, an AAVrh.74-based vector, or an AAVrh.10-based vector can provide the desired cardiac cell specificity.

Pharmaceutical compositions and kits

In one aspect, the present invention provides pharmaceutical compositions comprising an rAAV virion of the present invention and one or more pharmaceutically acceptable carriers, diluents or excipients.

For the purpose of administration, for example by injection, various solutions such as sterile aqueous solutions can be used. Such aqueous solutions can be buffered if necessary and the liquid diluent first rendered isotonic with saline or glucose. Solutions of rAAV as a free acid (DNA contains acidic phosphate groups) or a pharmacologically acceptable salt can be prepared in water suitably mixed with a surfactant such as poloxamer 188, for example, at a concentration of 0.001% or 0.01%. rAAV can also be dispersed in glycerol, liquid polyethylene glycols and mixtures thereof, and in oils. Under normal conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. In this regard, all sterile aqueous media used can be readily prepared by standard techniques well known to those skilled in the art.

The pharmaceutical forms suitable for injectable use include, but are not limited to, sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form is sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, a polyol (e.g., glycerol, propylene glycol, liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oils. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by maintaining the required particle size in the case of dispersion, and by the use of surfactants. The action of microorganisms can be prevented by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In many cases, it will be preferable to include isotonic agents, such as sugars or sodium chloride. Prolonged absorption of injection compositions can be achieved by using agents that slow absorption, such as aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating rAAV in the required amount in an appropriate solvent with various of the other ingredients enumerated above, if necessary, followed by filter sterilization. Generally, dispersions are prepared by incorporating the sterilized active ingredient into a sterile carrier medium which contains the basic dispersion medium and the required other ingredients from among those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and lyophilization techniques which yield a powder of the active ingredient plus any additional required ingredient from a previously filter sterilized solution thereof.

In another aspect, the present invention provides a kit comprising an rAAV virion of the present invention and instructions for use.

Methods of application

In one aspect, the present invention provides a method of increasing MLP activity in a cell, comprising contacting the cell with an rAAV of the present invention. In another aspect, the present invention provides a method of increasing MLP activity in a subject, comprising administering an rAAV of the present invention. In some embodiments, the cell and/or subject are deficient in messenger RNA expression levelsCSRP3or MLP protein and/or activity and/or have a loss-of-function mutation inCSRP3. The cell may be a heart cell, for example, cardiomyocyte.

In some embodiments, the method promotes the survival of a cardiac cell, such as cardiomyocyte, in cell culture and/or under conditionsin vivoIn some embodiments, the method stimulates and/or restores cardiac function.

Treatment methods

In another aspect, the present invention provides a method of treating a disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of an rAAV virion of the present invention. In some embodiments, the disease or disorder is a cardiac disease or disorder. Exemplary cardiac disorders include heart failure, hypertrophic cardiomyopathy, and dilated cardiomyopathy. In some embodiments, the subject has a genetic disorder of CSRP3 expression or function. In some embodiments, the disease or disorder is HCM or DCM. In some embodiments, the disease or disorder is hereditary hypertrophic cardiomyopathy-12 (CMH12). In some embodiments, the disease or disorder is dilated cardiomyopathy-1M (CMD1M). In some embodiments, the disease or disorder is skeletal myotrophy. In some embodiments, the disease or disorder is facioscapulohumeral muscular dystrophy, nemaline myopathy, or limb-girdle muscular dystrophy type 2B. In some embodiments, the disease or disorder is limb-girdle muscular dystrophy type 2A, Duchenne muscular dystrophy, or dermatomyositis.

AAV-mediated delivery of MLP protein to the heart may prolong survival, prevent or attenuate cardiac cell degeneration, heart failure, scarring, decreased ejection fraction, arrhythmia, angina, obstructive HCM or DCM, ventricular hypertrophy (IVS: range 14-32 mm), ventricular tachycardia, mild NYHA I-II, exercise intolerance, angina (chest pain), sudden cardiac death, myalgias, and exertional cramps.

The methods described herein can provide efficient biodistribution in the heart. They can result in robust expression in all or a significant proportion of cardiac cells, such as cardiomyocytes. Notably, the methods disclosed herein can provide for long-term expression of the MLP protein throughout the life of a subject following administration of the AAV-based vector.

The present invention also provides various types of combination therapies. In particular, combinations of the methods of the present invention with standard treatments (e.g., corticosteroids or drugs for local pressure reduction), as well as combinations with new treatments are provided. In some cases, the subject can be treated with a steroid and/or a combination of immunosuppressants to prevent or reduce the immune response to the administration of the rAAV described herein.

In some embodiments, the AAV-based vector is administered at a dose of about 1×10 ¹² to 5×10 ¹⁴ vector genomes (vg) of the AAV-based vector per kilogram (kg) of total body weight of the subject (vg/kg). In some embodiments, the AAV-based vector is administered at a dose of about 1×10 ¹³ to 5×10 ¹⁴ vg/kg. In some embodiments, the AAV-based vector is administered at a dose of about 5×10 ¹³ to 3×10 ¹⁴ vg/kg. In some embodiments, the AAV-based vector is administered at a dose of about 5×10 ¹³ to 1×10 ¹⁴ vg/kg. In some embodiments, the AAV vector is administered at a dose of less than about 1×10 ¹² vg/kg, less than about 3×10 ¹² vg/kg, less than about 5×10 ¹² vg/kg, less than about 7×10 ¹² vg/kg, less than about 1×10 ¹³ vg/kg, less than about 3×10 ¹³ vg/kg, less than about 5×10 ¹³ vg/kg, less than about 7×10 ¹³ vg/kg, less than about 1×10 ¹⁴ vg/kg, less than about 3×10 ¹⁴ vg/kg, less than about 5×10 ¹⁴ vg/kg, less than about 7×10 ¹⁴ vg/kg, less than about 1×10 ¹⁵ vg/kg, less than about 3×10 ¹⁵ vg/kg, less than approximately 5×10 ¹⁵ vg/kg, or less than approximately 7×10 ¹⁵ vg/kg.

In some embodiments, the AAV vector is administered at a dose of about 1×10 ¹² vg/kg, about 3×10 ¹² vg/kg, about 5×10 ¹² vg/kg, about 7×10 ¹² vg/kg, about 1×10 ¹³ vg/kg, about 3×10 ¹³ vg/kg, about 5×10 ¹³ vg/kg, about 7×10 ¹³ vg/kg, about 1×10 ¹⁴ vg/kg, about 3×10 ¹⁴ vg/kg, about 5×10 ¹⁴ vg/kg, about 7×10 ¹⁴ vg/kg, about 1×10 ¹⁵ vg/kg, about 3×10 ¹⁵ vg/kg, about 5×10 ¹⁵ vg/kg, or approximately 7×10 ¹⁵ vg/kg.

In some embodiments, the AAV vector is administered at a dose of 1×10 ¹² vg/kg, 3×10 ¹² vg/kg, 5×10 ¹² vg/kg, 7×10 ¹² vg/kg, 1×10 ¹³ vg/kg, 3×10 ¹³ vg/kg, 5×10 ¹³ vg/kg, 7×10 ¹³ vg/kg, 1×10 ¹⁴ vg/kg, 3×10 ¹⁴ vg/kg, 5×10 ¹⁴ vg/kg, 7×10 ¹⁴ vg/kg, 1×10 ¹⁵ vg/kg, 3×10 ¹⁵ vg/kg, 5×10 ¹⁵ vg/kg, or 7×10 ¹⁵ vg/kg.

In some embodiments, the AAV-based vector is administered systemically at a dose of about 1×10 ¹² to 5×10 ¹⁴ vector genomes (vg) of the AAV-based vector per kilogram (kg) of total body weight of the subject (vg/kg). In some embodiments, the AAV-based vector is administered systemically at a dose of about 1×10 ¹³ to 5×10 ¹⁴ vg/kg. In some embodiments, the AAV-based vector is administered systemically at a dose of about 5×10 ¹³ to 3×10 ¹⁴ vg/kg. In some embodiments, the AAV-based vector is administered systemically at a dose of about 5×10 ¹³ to 1×10 ¹⁴ vg/kg. In some embodiments, the AAV vector is administered systemically at a dose of less than about 1×10 ¹² vg/kg, less than about 3×10 ¹² vg/kg, less than about 5×10 ¹² vg/kg, less than about 7×10 ¹² vg/kg, less than about 1×10 ¹³ vg/kg, less than about 3×10 ¹³ vg/kg, less than about 5×10 ¹³ vg/kg, less than about 7×10 ¹³ vg/kg, less than about 1×10 ¹⁴ vg/kg, less than about 3×10 ¹⁴ vg/kg, less than about 5×10 ¹⁴ vg/kg, less than about 7×10 ¹⁴ vg/kg, less than about 1×10 ¹⁵ vg/kg, less than approximately 3×10 ¹⁵ wg/kg, less than approximately 5×10 ¹⁵ wg/kg, or less than approximately 7×10 ¹⁵ wg/kg.

In some embodiments, the AAV vector is administered systemically at a dose of about 1×10 ¹² vg/kg, about 3×10 ¹² vg/kg, about 5×10 ¹² vg/kg, about 7×10 ¹² vg/kg, about 1×10 ¹³ vg/kg, about 3×10 ¹³ vg/kg, about 5×10 ¹³ vg/kg, about 7×10 ¹³ vg/kg, about 1×10 ¹⁴ vg/kg, about 3×10 ¹⁴ vg/kg, about 5×10 ¹⁴ vg/kg, about 7×10 ¹⁴ vg/kg, about 1×10 ¹⁵ vg/kg, about 3×10 ¹⁵ vg/kg, about 5×10 ¹⁵ vg/kg or approximately 7×10 ¹⁵ vg/kg.

In some embodiments, the AAV vector is administered systemically at a dose of 1×10 ¹² vg/kg, 3×10 ¹² vg/kg, 5×10 ¹² vg/kg, 7×10 ¹² vg/kg, 1×10 ¹³ vg/kg, 3×10 ¹³ vg/kg, 5×10 ¹³ vg/kg, 7×10 ¹³ vg/kg, 1×10 ¹⁴ vg/kg, 3×10 ¹⁴ vg/kg, 5×10 ¹⁴ vg/kg, 7×10 ¹⁴ vg/kg, 1×10 ¹⁵ vg/kg, 3×10 ¹⁵ vg/kg, 5×10 ¹⁵ vg/kg, or 7×10 ¹⁵ vg/kg.

In some embodiments, the AAV-based vector is administered intravenously at a dose of about 1×10 ¹² to 5×10 ¹⁴ vector genomes (vg) of the AAV-based vector per kilogram (kg) of total body weight of the subject (vg/kg). In some embodiments, the AAV-based vector is administered intravenously at a dose of about 1×10 ¹³ to 5×10 ¹⁴ vg/kg. In some embodiments, the AAV-based vector is administered intravenously at a dose of about 5×10 ¹³ to 3×10 ¹⁴ vg/kg. In some embodiments, the AAV-based vector is administered intravenously at a dose of about 5×10 ¹³ to 1×10 ¹⁴ vg/kg. In some embodiments, the AAV vector is administered intravenously at a dose of less than about 1×10 ¹² vg/kg, less than about 3×10 ¹² vg/kg, less than about 5×10 ¹² vg/kg, less than about 7×10 ¹² vg/kg, less than about 1×10 ¹³ vg/kg, less than about 3×10 ¹³ vg/kg, less than about 5×10 ¹³ vg/kg, less than about 7×10 ¹³ vg/kg, less than about 1×10 ¹⁴ vg/kg, less than about 3×10 ¹⁴ vg/kg, less than about 5×10 ¹⁴ vg/kg, less than about 7×10 ¹⁴ vg/kg, less than about 1×10 ¹⁵ vg/kg, less than than approximately 3×10 ¹⁵ wg/kg, less than approximately 5×10 ¹⁵ wg/kg, or less than approximately 7×10 ¹⁵ wg/kg.

In some embodiments, the AAV vector is administered intravenously at a dose of about 1×10 ¹² vg/kg, about 3×10 ¹² vg/kg, about 5×10 ¹² vg/kg, about 7×10 ¹² vg/kg, about 1×10 ¹³ vg/kg, about 3×10 ¹³ vg/kg, about 5×10 ¹³ vg/kg, about 7×10 ¹³ vg/kg, about 1×10 ¹⁴ vg/kg, about 3×10 ¹⁴ vg/kg, about 5×10 ¹⁴ vg/kg, about 7×10 ¹⁴ vg/kg, about 1×10 ¹⁵ vg/kg, about 3×10 ¹⁵ vg/kg, about 5×10 ¹⁵ vg/kg or approximately 7×10 ¹⁵ vg/kg.

In some embodiments, the AAV vector is administered intravenously at a dose of 1×10 ¹² vg/kg, 3×10 ¹² vg/kg, 5×10 ¹² vg/kg, 7×10 ¹² vg/kg, 1×10 ¹³ vg/kg, 3×10 ¹³ vg/kg, 5×10 ¹³ vg/kg, 7×10 ¹³ vg/kg, 1×10 ¹⁴ vg/kg, 3×10 ¹⁴ vg/kg, 5×10 ¹⁴ vg/kg, 7×10 ¹⁴ vg/kg, 1×10 ¹⁵ vg/kg, 3×10 ¹⁵ vg/kg, 5×10 ¹⁵ vg/kg, or 7×10 ¹⁵ vg/kg.

Evidence of functional improvement, clinical benefit, or efficacy in patients may be demonstrated by changes in New York Heart Association functional classification (NYHA class), abnormal electrocardiogram, left ventricular end-diastolic/end-systolic diameter, maximal interventricular wall thickness, maximal posterior wall thickness, E-wave and A-wave frequency, peak early and peak late transmitral filling velocity, early diastolic and late diastolic tissue Doppler velocity, arterial hypertension, and degree of cardiac hypertrophy. Additional myocardial histology will identify an AAV-mediated beneficial effect of MLP as demonstrated by reduced cardiomyocyte hypertrophy, reduced myocyte bulk, and reduced interstitial and perivascular fibrosis and scarring compared with baseline or disease-matched controls.

Introduction of compositions

The administration of an effective dose of the compositions can be carried out by standard routes in the art, including, but not limited to, systemic, local, direct injection, intravenous, intracardiac administration. In some cases, the administration includes systemic, local, direct injection, intravenous, intracardiac injection. The administration can be carried out by cardiac catheterization.

In some embodiments, the present invention provides for local administration and systemic administration of an effective dose of rAAV and the compositions of the present invention. For example, systemic administration can be administration into the circulatory system such that the entire body is exposed. Systemic administration includes parenteral administration via injection, infusion, or implantation. Routes of administration of the compositions disclosed herein include intravenous ("IV") administration, intraperitoneal ("IP") administration, intramuscular ("IM") administration, intralesional or subcutaneous ("SC") administration, or implantation of a slow release device, such as mini-osmotic pump, depot formulation, etc. In some embodiments, the methods of the present invention comprise administering the AAV vector of the present invention or a pharmaceutical composition comprising the same via intravenous, intramuscular, intraarterial, intrarenal, intraurethral, intracardiac, intracoronary, intramyocardial, intradermal, epidural, subcutaneous, intraperitoneal, intraventricular, iontophoretic, or intracranial administration.

In particular, the rAAV of the present invention may be administered by any physical method that allows the recombinant rAAV vector to be transported into the target tissue of the animal. Administration includes, but is not limited to, cardiac injection.

In some embodiments, the methods of the present invention involve intracardiac delivery. Infusion can be performed using a special cannula, catheter, syringe/needle, using an infusion pump. Administration can involve delivering an effective amount of rAAV virion or a pharmaceutical composition comprising rAAV virion to the heart. This can be accomplished, for example, by intravenous, intramuscular, intraarterial, intrarenal, intraurethral, intracardiac, intracoronary, intramyocardial, intradermal, epidural, subcutaneous, intraperitoneal, intraventricular, iontophoretic or intracranial administration. The compositions of the present invention can additionally be administered intravenously.

The method of treatment disclosed herein can reduce and/or prevent one or more symptoms, including but not limited to ventricular hypertrophy, ventricular tachycardia, moderate NYHA I-II, exercise intolerance, and angina.

EXAMPLES

Example 1. Preclinical bioactivity and efficacy

The vectors illustrated in Fig. 1-4 are tested under the conditionsin vitrousing cultured cardiomyocytes (eg, cardiomyocytes with induced pluripotent stem cells, iPSC-CM). MLP expression is assessed by immunofluorescence and Western blotting. Phosphorylation assays reveal a decrease in protein kinase C-alpha (PKC-A) autophosphorylation.

The selected vectors are tested under the following conditions:in vivousing MLP-deficient or MLP-mutant mouse models of cardiomyopathy (e.g., models with C58G inclusion (KI) or models with KI W4R). Efficacy is determined by measuring left ventricular ejection fraction (LVEF) and/or left ventricular end-diastolic dimension (LVED) using echocardiography, reduction in total cardiac mass (e.g. normalized by tibial length), invasive hemodynamic assessments of left ventricular function on dP/dt_max, dP/dt_minand Tau relaxation constants or reduction in left and/or right ventricular hypertrophy as assessed histologically. Additionally, efficacy in conditionsin vivoin a mouse model, biomarkers are measured, including but not limited to atrial natriuretic factor (Nppa) gene expression, brain natriuretic peptide (Nppb) gene expression, and beta-myosin heavy chain protein expression. Physiological efficacy is determined by testing protein kinase C-alpha (PKC-A) activity, cardiac MLP phosphorylation, and ubiquitin-dependent proteasome degradation. In the case of AAV-based vectors, normalization or amelioration of the condition is observed in response to treatment.

Example 2. Protein expression in human cardiomyocytes

The vectors illustrated in Fig. 1-4 were tested in vitro using a control cell line (CHO-Lec2; Fig. 5A ) and cultured cardiomyocytes (differentiated cell line AC16; Fig. 5B ). Expression of muscle LIM protein (MLP; the protein encoded by CSRP3 ) was assessed by Western blotting.

Figure 5A shows CSRP3 expression in transduced CHO-Lec2. Figure 5B shows CSRP3 expression in transduced cardiomyocytes (differentiated cell line AC16 - Sigma-Aldrich®, Cat. No. SCC109). Cells were transduced with an MOI of 3E5 from each vector; after 6 days, cell lysates were collected and Western blotted using a polyclonal antibody to CSRP3 (Thermo-Fisher® PA5-29155 1:1000).

The expression of MLP protein of the CSRP3 transgene is higher with the MHCK7 promoter than with the hTNNT2 ("hTnT") promoter. Both AAV9 and AAVrh74 serotypes of the AAV-based vector are able to transduce the cardiomyocyte cell line. Based on the data shown in Fig. 5B, the expression of MLP protein is clearly higher with the AAVrh74-based vector than with the AAV9-based vector.

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SEQUENCE LIST

<110> Spacecraft Seven, LLSI

<120> GENE THERAPY BASED ON CSRP3 (RICH IN CYSTEINE AND GLYCINE

SQUIRREL 3)

<130> ROPA-020/01WO 329592-22266

<150> US 63/061727

<151> 2020-08-05

<160> 86

<170> PatentIn version 3.5

<210> 1

<211> 194

<212> PROTEIN

<213> Homo sapiens

<400> 1

Met Pro Asn Trp Gly Gly Gly Ala Lys Cys Gly Ala Cys Glu Lys Thr

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Val Tyr His Ala Glu Glu Ile Gln Cys Asn Gly Arg Ser Phe His Lys

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Thr Cys Phe His Cys Met Ala Cys Arg Lys Ala Leu Asp Ser Thr Thr

35 40 45

Val Ala Ala His Glu Ser Glu Ile Tyr Cys Lys Val Cys Tyr Gly Arg

50 55 60

Arg Tyr Gly Pro Lys Gly Ile Gly Tyr Gly Gln Gly Ala Gly Cys Leu

65 70 75 80

Ser Thr Asp Thr Gly Glu His Leu Gly Leu Gln Phe Gln Gln Ser Pro

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Lys Pro Ala Arg Ser Val Thr Thr Ser Asn Pro Ser Lys Phe Thr Ala

100 105 110

Lys Phe Gly Glu Ser Glu Lys Cys Pro Arg Cys Gly Lys Ser Val Tyr

115 120 125

Ala Ala Glu Lys Val Met Gly Gly Gly Lys Pro Trp His Lys Thr Cys

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Phe Arg Cys Ala Ile Cys Gly Lys Ser Leu Glu Ser Thr Asn Val Thr

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Asp Lys Asp Gly Glu Leu Tyr Cys Lys Val Cys Tyr Ala Lys Asn Phe

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Gly Pro Thr Gly Ile Gly Phe Gly Gly Leu Thr Gln Gln Val Glu Lys

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Lys Glu

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Met Pro Asn Trp Gly Gly Gly Ala Lys Cys Gly Ala Cys Glu Lys Thr

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Val Tyr His Ala Glu Glu Ile Gln Cys Asn Gly Arg Ser Phe His Lys

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Thr Cys Phe His Cys Ser Pro Gln Ser Arg His Ala Gln Leu Pro Pro

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Ala Thr Leu Pro Asn Ser Leu Arg Ser Leu Glu Ser Pro Arg Ser Ala

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Leu Asp Val Ala Ser Gln Ser Met Leu Leu Arg Arg Leu Trp Glu Val

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Ala Ser Leu Gly Thr Arg Pro Val Ser Ala Val Pro Ser Val Gly Arg

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Val Trp Ser Pro Gln Met Ser Leu Thr Lys Met Gly Asn Phe Ile Ala

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Lys Phe Ala Met Pro Lys Ile Leu Ala Pro Arg Val Leu Gly Leu Glu

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Ala Leu His Asn Lys Trp Lys Arg Lys Asn Glu Glu Val Arg Arg Phe

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<212> DNA

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atgccaaact ggggcggagg cgcaaaatgt ggagcctgtg aaaagaccgt ctaccatgca 60

gaagaaatcc agtgcaatgg aaggagtttc cacaagacgt gtttccactg catggcctgc 120

aggaaggctc ttgacagcac gacagtcgcg gctcatgagt cggagatcta ctgcaaggtg 180

tgctatgggc gcagatatgg ccccaaaggg atcgggtatg gacaaggcgc tggctgtctc 240

agcacagaca cgggcgagca tctcggcctg cagttccaac agtccccaaa gccggcacgc 300

tcagttacca ccagcaaccc ttccaaattc actgcgaagt ttggagagtc cgagaagtgc 360

cctcgatgtg gcaagtcagt ctatgctgct gagaaggtta tgggaggtgg caagccttgg 420

cacaagacct gtttccgctg tgccatctgt gggaagagtc tggagtccac aaatgtcact 480

gacaaagatg gggaacttta ttgcaaagtt tgctatgcca aaaattttgg ccccacgggt 540

attgggtttg gaggccttac acaacaagtg gaaaagaaag aa 582

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gccaccatgg 10

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<223> Polynucleotide encoding MLP with Kozak sequence

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gccaccatgc caaactgggg cggaggcgca aaatgtggag cctgtgaaaa gaccgtctac 60

catgcagaag aaatccagtg caatggaagg agtttccaca agacgtgttt ccactgcatg 120

gcctgcagga aggctcttga cagcacgaca gtcgcggctc atgagtcgga gatctactgc 180

aaggtgtgct atgggcgcag atatggcccc aaagggatcg ggtatggaca aggcgctggc 240

tgtctcagca cagacacggg cgagcatctc ggcctgcagt tccaacagtc cccaaagccg 300

gcacgctcag ttaccaccag caacccttcc aaattcactg cgaagtttgg agagtccgag 360

aagtgccctc gatgtggcaa gtcagtctat gctgctgaga aggttatggg aggtggcaag 420

ccttggcaca agacctgttt ccgctgtgcc atctgtggga agagtctgga gtccacaaat 480

gtcactgaca aagatgggga actttattgc aaagtttgct atgccaaaaa ttttggcccc 540

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acccttcaga ttaaaaataa ctgaggtaag ggcctgggta ggggaggtgg tgtgagacgc 60

tcctgtctct cctctatctg cccatcggcc ctttggggag gaggaatgtg cccaaggact 120

aaaaaaaggc catggagcca gaggggcgag ggcaacagac ctttcatggg caaaccttgg 180

ggccctgctg tctagcatgc cccactacgg gtctaggctg cccatgtaag gaggcaaggc 240

ctggggacac ccgagatgcc tggttataat taacccagac atgtggctgc cccccccccc 300

ccaacacctg ctgcctctaa aaataaccct gtccctggtg gatcccctgc atgcgaagat 360

cttcgaacaa ggctgtgggg gactgagggc aggctgtaac aggcttgggg gccagggctt 420

atacgtgcct gggactccca aagtattact gttccatgtt cccggcgaag ggccagctgt 480

cccccgccag ctagactcag cacttagttt aggaaccagt gagcaagtca gcccttgggg 540

cagcccatac aaggccatgg ggctgggcaa gctgcacgcc tgggtccggg gtgggcacgg 600

tgcccgggca acgagctgaa agctcatctg ctctcagggg cccctccctg gggacagccc 660

ctcctggcta gtcacaccct gtaggctcct ctatataacc caggggcaca ggggctgccc 720

tcattctacc accacctcca cagcacagac agacactcag gagccagcca ggccaccatg 780

ccaaactggg gcggaggcgc aaaatgtgga gcctgtgaaa agaccgtcta ccatgcagaa 840

gaaatccagt gcaatggaag gagtttccac aagacgtgtt tccactgcat ggcctgcagg 900

aaggctcttg acagcacgac agtcgcggct catgagtcgg agatctactg caaggtgtgc 960

tatgggcgca gatatggccc caaagggatc gggtatggac aaggcgctgg ctgtctcagc 1020

acagacacgg gcgagcatct cggcctgcag ttccaacagt ccccaaagcc ggcacgctca 1080

gttaccacca gcaacccttc caaattcact gcgaagtttg gagagtccga gaagtgccct 1140

cgatgtggca agtcagtcta tgctgctgag aaggttatgg gaggtggcaa gccttggcac 1200

aagacctgtt tccgctgtgc catctgtggg aagagtctgg agtccacaaa tgtcactgac 1260

aaagatgggg aactttattg caaagtttgc tatgccaaaa attttggccc cacgggtatt 1320

gggtttggag gccttacaca acaagtggaa aagaaagaat gatcaacctc tggattacaa 1380

aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata 1440

cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc 1500

cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg 1560

tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac 1620

ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat 1680

cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt 1740

ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat 1800

tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc 1860

ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag 1920

tcggatctcc ctttgggccg cctccccgca ctgcccgggt ggcatccctg tgacccctcc 1980

ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata 2040

aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg 2100

ggggtggtat ggagcaaggg gcccaagttg ggaagaaacc tgtagggcct gc 2152

<210> 9

<211> 1925

<212> DNA

<213> Artificial sequence

<220>

<223> Lab-Made – hTnT-CSRP3 Expression Cassette

<400> 9

ctcagtccat taggagccag tagcctggaa gatgtcttta cccccagcat cagttcaagt 60

ggagcagcac ataactcttg ccctctgcct tccaagattc tggtgctgag acttatggag 120

tgtcttggag gttgccttct gccccccaac cctgctccca gctggccctc ccaggcctgg 180

gttgctggcc tctgctttat caggattctc aagagggaca gctggtttat gttgcatgac 240

tgttccctgc atatctgctc tggttttaaa tagcttatct gagcagctgg aggaccacat 300

gggcttatat ggcgtggggt acatgttcct gtagccttgt ccctggcacc tgccaaaata 360

gcagccaaca ccccccaccc ccaccgccat ccccctgccc cacccgtccc ctgtcgcaca 420

ttcctccctc cgcagggctg gctcaccagg ccccagccca catgcctgct taaagccctc 480

tccatcctct gcctcaccca gtccccgctg agactgagca gacgcctcca ggatctgtcg 540

gcaggccacc atgccaaact ggggcggagg cgcaaaatgt ggagcctgtg aaaagaccgt 600

ctaccatgca gaagaaatcc agtgcaatgg aaggagtttc cacaagacgt gtttccactg 660

catggcctgc aggaaggctc ttgacagcac gacagtcgcg gctcatgagt cggagatcta 720

ctgcaaggtg tgctatgggc gcagatatgg ccccaaaggg atcgggtatg gacaaggcgc 780

tggctgtctc agcacagaca cgggcgagca tctcggcctg cagttccaac agtccccaaa 840

gccggcacgc tcagttacca ccagcaaccc ttccaaattc actgcgaagt ttggagagtc 900

cgagaagtgc cctcgatgtg gcaagtcagt ctatgctgct gagaaggtta tgggaggtgg 960

caagccttgg cacaagacct gtttccgctg tgccatctgt gggaagagtc tggagtccac 1020

aaatgtcact gacaaagatg gggaacttta ttgcaaagtt tgctatgcca aaaattttgg 1080

ccccacgggt attgggtttg gaggccttac acaacaagtg gaaaagaaag aatgatcaac 1140

ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta 1200

cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt 1260

tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg 1320

ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg 1380

gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca 1440

cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca 1500

ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg 1560

ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag 1620

cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc 1680

gccctcagac gagtcggatc tccctttggg ccgcctcccc gcactgcccg ggtggcatcc 1740

ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag 1800

ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat 1860

tatggggtgg aggggggtgg tatggagcaa ggggcccaag ttgggaagaa acctgtaggg 1920

cctgc 1925

<210> 10

<211> 2152

<212> DNA

<213> Artificial sequence

<220>

<223> Lab Made – MHCK7-CSRP3 Expression Cassette

<400> 10

acccttcaga ttaaaaataa ctgaggtaag ggcctgggta ggggaggtgg tgtgagacgc 60

tcctgtctct cctctatctg cccatcggcc ctttggggag gaggaatgtg cccaaggact 120

aaaaaaaggc catggagcca gaggggcgag ggcaacagac ctttcatggg caaaccttgg 180

ggccctgctg tctagcatgc cccactacgg gtctaggctg cccatgtaag gaggcaaggc 240

ctggggacac ccgagatgcc tggttataat taacccagac atgtggctgc cccccccccc 300

ccaacacctg ctgcctctaa aaataaccct gtccctggtg gatcccctgc atgcgaagat 360

cttcgaacaa ggctgtgggg gactgagggc aggctgtaac aggcttgggg gccagggctt 420

atacgtgcct gggactccca aagtattact gttccatgtt cccggcgaag ggccagctgt 480

cccccgccag ctagactcag cacttagttt aggaaccagt gagcaagtca gcccttgggg 540

cagcccatac aaggccatgg ggctgggcaa gctgcacgcc tgggtccggg gtgggcacgg 600

tgcccgggca acgagctgaa agctcatctg ctctcagggg cccctccctg gggacagccc 660

ctcctggcta gtcacaccct gtaggctcct ctatataacc caggggcaca ggggctgccc 720

tcattctacc accacctcca cagcacagac agacactcag gagccagcca ggccaccatg 780

cccaattggg gtggagggagc taaatgtgga gcttgtgaaa aaacagttta tcatgctgaa 840

gaaattcaat gtaatggaag atcttttcat aaaacatgtt ttcattgtat ggcttgtaga 900

aaagcacttg attctacaac tgttgcagca catgaaagtg aaatctattg taaagtatgt 960

tatggaagaa gatatggacc aaaaggaatt ggatatggac aaggagcagg atgtctttct 1020

acagatactg gagaacattt gggattgcaa tttcaacaaa gtcctaaacc agctagatct 1080

gttacaacaa gtaatccatc aaaatttact gctaaatttg gagaatccga aaaatgtcct 1140

agatgtggaa aatcagtata tgctgctgaa aaagttatgg gaggtggaaa accatggcat 1200

aagacatgtt ttagatgtgc aatttgtggt aaatctttgg aatctacaaa tgttacagat 1260

aaagatggag aattgtattg taaagtttgt tatgctaaaa attttggacc tacaggtata 1320

ggatttggag gtttgacaca acaagttgaa aaaaaagaat gatcaacctc tggattacaa 1380

aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata 1440

cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc 1500

cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg 1560

tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac 1620

ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat 1680

cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt 1740

ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat 1800

tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc 1860

ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag 1920

tcggatctcc ctttgggccg cctccccgca ctgcccgggt ggcatccctg tgacccctcc 1980

ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata 2040

aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg 2100

ggggtggtat ggagcaaggg gcccaagttg ggaagaaacc tgtagggcct gc 2152

<210> 11

<211> 1925

<212> DNA

<213> Artificial sequence

<220>

<223> Lab Made - hTnT-CSRP3 Codon Optimized Cassette

expressions

<400> 11

ctcagtccat taggagccag tagcctggaa gatgtcttta cccccagcat cagttcaagt 60

ggagcagcac ataactcttg ccctctgcct tccaagattc tggtgctgag acttatggag 120

tgtcttggag gttgccttct gccccccaac cctgctccca gctggccctc ccaggcctgg 180

gttgctggcc tctgctttat caggattctc aagagggaca gctggtttat gttgcatgac 240

tgttccctgc atatctgctc tggttttaaa tagcttatct gagcagctgg aggaccacat 300

gggcttatat ggcgtggggt acatgttcct gtagccttgt ccctggcacc tgccaaaata 360

gcagccaaca ccccccaccc ccaccgccat ccccctgccc cacccgtccc ctgtcgcaca 420

ttcctccctc cgcagggctg gctcaccagg ccccagccca catgcctgct taaagccctc 480

tccatcctct gcctcaccca gtccccgctg agactgagca gacgcctcca ggatctgtcg 540

gcaggccacc atgcccaatt ggggtggagg agctaaatgt ggagcttgtg aaaaaacagt 600

ttatcatgct gaagaaattc aatgtaatgg aagatctttt cataaaacat gttttcattg 660

tatggcttgt agaaaagcac ttgattctac aactgttgca gcacatgaaa gtgaaatcta 720

ttgtaaagta tgttatggaa gaagatatgg accaaaagga attggatatg gacaaggagc 780

aggatgtctt tctacagata ctggagaaca tttgggattg caatttcaac aaagtcctaa 840

accagctaga tctgttacaa caagtaatcc atcaaaattt actgctaaat ttggagaatc 900

cgaaaaatgt cctagatgtg gaaaatcagt atatgctgct gaaaaagtta tgggaggtgg 960

aaaaccatgg cataagacat gttttagatg tgcaatttgt ggtaaatctt tggaatctac 1020

aaatgttaca gataaagatg gagaattgta ttgtaaagtt tgttatgcta aaaattttgg 1080

acctacaggt ataggatttg gaggtttgac acaacaagtt gaaaaaaaag aatgatcaac 1140

ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta 1200

cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt 1260

tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg 1320

ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg 1380

gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca 1440

cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca 1500

ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg 1560

ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag 1620

cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc 1680

gccctcagac gagtcggatc tccctttggg ccgcctcccc gcactgcccg ggtggcatcc 1740

ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag 1800

ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat 1860

tatggggtgg aggggggtgg tatggagcaa ggggcccaag ttgggaagaa acctgtaggg 1920

cctgc 1925

<210> 12

<211> 2430

<212> DNA

<213> Artificial sequence

<220>

<223> Made in the Lab – Complete Polynucleotide Sequence

vector genome

<400> 12

ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg 60

cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg 120

gccaactcca tcactagggg ttcctaccct tcagattaaa aataactgag gtaagggcct 180

gggtagggga ggtggtgtga gacgctcctg tctctcctct atctgcccat cggccctttg 240

gggaggagga atgtgcccaa ggactaaaaa aaggccatgg agccagaggg gcgagggcaa 300

cagacctttc atgggcaaac cttggggccc tgctgtctag catgccccac tacgggtcta 360

ggctgcccat gtaaggaggc aaggcctggg gacacccgag atgcctggtt ataattaacc 420

cagacatgtg gctgcccccc cccccccaac acctgctgcc tctaaaaata accctgtccc 480

tggtggatcc cctgcatgcg aagatcttcg aacaaggctg tgggggactg agggcaggct 540

gtaacaggct tggggggccag ggcttatacg tgcctgggac tcccaaagta ttactgttcc 600

atgttcccgg cgaagggcca gctgtccccc gccagctaga ctcagcactt agtttaggaa 660

ccagtgagca agtcagccct tggggcagcc catacaaggc catggggctg ggcaagctgc 720

acgcctgggt ccggggtggg cacggtgccc gggcaacgag ctgaaagctc atctgctctc 780

aggggcccct ccctggggac agcccctcct ggctagtcac accctgtagg ctcctctata 840

taacccaggg gcacaggggc tgccctcatt ctaccaccac ctccacagca cagacagaca 900

ctcaggagcc agccaggcca ccatgccaaa ctggggcgga ggcgcaaaat gtggagcctg 960

tgaaaagacc gtctaccatg cagaagaaat ccagtgcaat ggaaggagtt tccacaagac 1020

gtgtttccac tgcatggcct gcaggaaggc tcttgacagc acgacagtcg cggctcatga 1080

gtcggagatc tactgcaagg tgtgctatgg gcgcagatat ggccccaaag ggatcgggta 1140

tggacaaggc gctggctgtc tcagcacaga cacgggcgag catctcggcc tgcagttcca 1200

acagtcccca aagccggcac gctcagttac caccagcaac ccttccaaat tcactgcgaa 1260

gtttggagag tccgagaagt gccctcgatg tggcaagtca gtctatgctg ctgagaaggt 1320

tatgggaggt ggcaagcctt ggcacaagac ctgtttccgc tgtgccatct gtgggaagag 1380

tctggagtcc acaaatgtca ctgacaaaga tggggaactt tattgcaaag tttgctatgc 1440

caaaaatttt ggccccacgg gtattgggtt tggaggcctt acacaacaag tggaaaagaa 1500

agaatgatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 1560

ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 1620

cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 1680

agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 1740

ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 1800

tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 1860

ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc 1920

tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 1980

ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 2040

gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcactgcc 2100

cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc 2160

agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc 2220

cttctataat attatggggt ggaggggggt ggtatggagc aaggggccca agttgggaag 2280

aaacctgtag ggcctgcagg aacccctagt gatggagact ccctctctgc gcgctcgctc 2340

gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc 2400

agtgagcgag cgagcgcgca gagagggagt 2430

<210> 13

<211> 2203

<212> DNA

<213> Artificial sequence

<220>

<223> Made in the Lab - Complete Polynucleotide Sequence

vector genome

<400> 13

ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg 60

cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg 120

gccaactcca tcactagggg ttcctctcag tccattagga gccagtagcc tggaagatgt 180

ctttaccccc agcatcagtt caagtggagc agcacataac tcttgccctc tgccttccaa 240

gattctggtg ctgagactta tggagtgtct tggaggttgc cttctgcccc ccaaccctgc 300

tcccagctgg ccctcccagg cctgggttgc tggcctctgc tttatcagga ttctcaagag 360

ggacagctgg tttatgttgc atgactgttc cctgcatatc tgctctggtt ttaaatagct 420

tatctgagca gctggaggac cacatgggct tatatggcgt ggggtacatg ttcctgtagc 480

cttgtccctg gcacctgcca aaatagcagc caacaccccc cacccccacc gccatccccc 540

tgccccaccc gtcccctgtc gcacattcct ccctccgcag ggctggctca ccaggcccca 600

gcccacatgc ctgcttaaag ccctctccat cctctgcctc acccagtccc cgctgagact 660

gagcagacgc ctccaggatc tgtcggcagg ccaccatgcc aaactggggc ggaggcgcaa 720

aatgtggagc ctgtgaaaag accgtctacc atgcagaaga aatccagtgc aatggaagga 780

gtttccacaa gacgtgtttc cactgcatgg cctgcaggaa ggctcttgac agcacgacag 840

tcgcggctca tgagtcggag atctactgca aggtgtgcta tgggcgcaga tatggcccca 900

aagggatcgg gtatggacaa ggcgctggct gtctcagcac agacacgggc gagcatctcg 960

gcctgcagtt ccaacagtcc ccaaagccgg cacgctcagt taccaccagc aacccttcca 1020

aattcactgc gaagtttgga gagtccgaga agtgccctcg atgtggcaag tcagtctatg 1080

ctgctgagaa ggttatggga ggtggcaagc cttggcacaa gacctgtttc cgctgtgcca 1140

tctgtgggaa gagtctggag tccacaaatg tcactgacaa agatggggaa ctttattgca 1200

aagtttgcta tgccaaaaat tttggcccca cgggtattgg gtttggaggc cttacacaac 1260

aagtggaaaa gaaagaatga tcaacctctg gattacaaaa tttgtgaaag attgactggt 1320

attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 1380

catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 1440

tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 1500

gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 1560

ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 1620

tggacaggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg 1680

tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 1740

tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 1800

cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 1860

tccccgcact gcccgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg 1920

aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt 1980

ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc 2040

ccaagttggg aagaaacctg tagggcctgc aggaacccct agtgatggag actccctctc 2100

tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg 2160

cccggggcggc ctcagtgagc gagcgagcgc gcagagaggg agt 2203

<210> 14

<211> 2430

<212> DNA

<213> Artificial sequence

<220>

<223> Made in the Lab - Complete Polynucleotide Sequence

vector genome

<400> 14

ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg 60

cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg 120

gccaactcca tcactagggg ttcctaccct tcagattaaa aataactgag gtaagggcct 180

gggtagggga ggtggtgtga gacgctcctg tctctcctct atctgcccat cggccctttg 240

gggaggagga atgtgcccaa ggactaaaaa aaggccatgg agccagaggg gcgagggcaa 300

cagacctttc atgggcaaac cttggggccc tgctgtctag catgccccac tacgggtcta 360

ggctgcccat gtaaggaggc aaggcctggg gacacccgag atgcctggtt ataattaacc 420

cagacatgtg gctgcccccc cccccccaac acctgctgcc tctaaaaata accctgtccc 480

tggtggatcc cctgcatgcg aagatcttcg aacaaggctg tgggggactg agggcaggct 540

gtaacaggct tggggggccag ggcttatacg tgcctgggac tcccaaagta ttactgttcc 600

atgttcccgg cgaagggcca gctgtccccc gccagctaga ctcagcactt agtttaggaa 660

ccagtgagca agtcagccct tggggcagcc catacaaggc catggggctg ggcaagctgc 720

acgcctgggt ccggggtggg cacggtgccc gggcaacgag ctgaaagctc atctgctctc 780

aggggcccct ccctggggac agcccctcct ggctagtcac accctgtagg ctcctctata 840

taacccaggg gcacaggggc tgccctcatt ctaccaccac ctccacagca cagacagaca 900

ctcaggagcc agccaggcca ccatgcccaa ttggggtgga ggagctaaat gtggagcttg 960

tgaaaaaaca gtttatcatg ctgaagaaat tcaatgtaat ggaagatctt ttcataaaac 1020

atgttttcat tgtatggctt gtagaaaagc acttgattct acaactgttg cagcacatga 1080

aagtgaaatc tattgtaaag tatgttatgg aagaagatat ggaccaaaag gaattggata 1140

tggacaagga gcaggatgtc tttctacaga tactggagaa catttgggat tgcaatttca 1200

acaaagtcct aaaccagcta gatctgttac aacaagtaat ccatcaaaat ttactgctaa 1260

atttggagaa tccgaaaaat gtcctagatg tggaaaatca gtatatgctg ctgaaaaagt 1320

tatgggaggt ggaaaaccat ggcataagac atgttttaga tgtgcaattt gtggtaaatc 1380

tttggaatct acaaatgtta cagataaaga tggagaattg tattgtaaag tttgttatgc 1440

taaaaatttt ggacctacag gtataggatt tggaggtttg acacaacaag ttgaaaaaaa 1500

agaatgatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 1560

ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 1620

cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 1680

agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 1740

ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 1800

tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 1860

ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc 1920

tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 1980

ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 2040

gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcactgcc 2100

cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc 2160

agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc 2220

cttctataat attatggggt ggaggggggt ggtatggagc aaggggccca agttgggaag 2280

aaacctgtag ggcctgcagg aacccctagt gatggagact ccctctctgc gcgctcgctc 2340

gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc 2400

agtgagcgag cgagcgcgca gagagggagt 2430

<210> 15

<211> 2203

<212> DNA

<213> Artificial sequence

<220>

<223> Made in the Lab - Complete Polynucleotide Sequence

vector genome

<400> 15

ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg 60

cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg 120

gccaactcca tcactagggg ttcctctcag tccattagga gccagtagcc tggaagatgt 180

ctttaccccc agcatcagtt caagtggagc agcacataac tcttgccctc tgccttccaa 240

gattctggtg ctgagactta tggagtgtct tggaggttgc cttctgcccc ccaaccctgc 300

tcccagctgg ccctcccagg cctgggttgc tggcctctgc tttatcagga ttctcaagag 360

ggacagctgg tttatgttgc atgactgttc cctgcatatc tgctctggtt ttaaatagct 420

tatctgagca gctggaggac cacatgggct tatatggcgt ggggtacatg ttcctgtagc 480

cttgtccctg gcacctgcca aaatagcagc caacaccccc cacccccacc gccatccccc 540

tgccccaccc gtcccctgtc gcacattcct ccctccgcag ggctggctca ccaggcccca 600

gcccacatgc ctgcttaaag ccctctccat cctctgcctc acccagtccc cgctgagact 660

gagcagacgc ctccaggatc tgtcggcagg cccacatgcc caattggggt ggaggagcta 720

aatgtggagc ttgtgaaaaa acagtttatc atgctgaaga aattcaatgt aatggaagat 780

cttttcataa aacatgtttt cattgtatgg cttgtagaaa agcacttgat tctacaactg 840

ttgcagcaca tgaaagtgaa atctattgta aagtatgtta tggaagaaga tatggaccaa 900

aaggaattgg atatggacaa ggagcaggat gtctttctac agatactgga gaacatttgg 960

gattgcaatt tcaacaaagt cctaaaccag ctagatctgt tacaacaagt aatccatcaa 1020

aatttactgc taaatttgga gaatccgaaa aatgtcctag atgtggaaaa tcagtatatg 1080

ctgctgaaaa agttatggga ggtggaaaac catggcataa gacatgtttt agatgtgcaa 1140

tttgtggtaa atctttggaa tctacaaatg ttacagataa agatggagaa ttgtattgta 1200

aagtttgtta tgctaaaaat tttggaccta caggtatagg atttggaggt ttgacacaac 1260

aagttgaaaa aaaagaatga tcaacctctg gattacaaaa tttgtgaaag attgactggt 1320

attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 1380

catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 1440

tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 1500

gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 1560

ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 1620

tggacaggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg 1680

tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 1740

tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 1800

cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 1860

tccccgcact gcccgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg 1920

aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt 1980

ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc 2040

ccaagttggg aagaaacctg tagggcctgc aggaacccct agtgatggag actccctctc 2100

tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg 2160

cccgggcggc ctcagtgagc gagcgagcgc gcagagaggg agt 2203

<210> 16

<211> 13

<212> DNA

<213> Artificial sequence

<220>

<223> Kozak sequence motif

<400> 16

gccgccrcca ugg 13

<210> 17

<211> 13

<212> DNA

<213> Artificial sequence

<220>

<223> Kozak sequence motif

<400> 17

gccgccrcca ugc 13

<210> 18

<211> 10

<212> DNA

<213> Artificial sequence

<220>

<223> Kozak sequence motif

<400> 18

gacaccaugg 10

<210> 19

<211> 10

<212> DNA

<213> Artificial sequence

<220>

<223> Kozak sequence motif

<400> 19

gacaccaugc 10

<210> 20

<211> 141

<212> DNA

<213> Adeno-associated virus

<400> 20

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc t 141

<210> 21

<211> 168

<212> DNA

<213> Adeno-associated virus 2

<400> 21

gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 60

tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag 120

gggttccttg tagttaatga ttaacccgcc atgctactta tctacgta 168

<210> 22

<211> 170

<212> DNA

<213> Adeno-associated virus

<400> 22

ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt 60

ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 120

aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta 170

<210> 23

<211> 145

<212> DNA

<213> Adeno-associated virus

<400> 23

ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg 60

cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg 120

gccaactcca tcactagggg ttcct 145

<210> 24

<211> 141

<212> DNA

<213> Adeno-associated virus

<400> 24

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 120

gagcgcgcag ctgcctgcag g 141

<210> 25

<211> 168

<212> DNA

<213> Adeno-associated virus 2

<400> 25

tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag 60

ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc 120

cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgc 168

<210> 26

<211> 133

<212> DNA

<213> Adeno-associated virus 2

<400> 26

aggaacccct agtgatggag actccctctc tgcgcgctcg ctcgctcact gaggccgggc 60

gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc 120

gcagaggg agt 133

<210> 27

<211> 124

<212> DNA

<213> Artificial sequence

<220>

<223> Made in the Lab - Vector Filler Sequence <400> 27

gcggcaattc agtcgataac tataacggtc ctaaggtagc gatttaaata cgcgctctct 60

taaggtagcc ccgggacgcg tcaattgact acaaaccgag tatctgcaga gggccctgcg 120

tag 124

<210> 28

<211> 84

<212> DNA

<213> Artificial sequence

<220>

<223> Made in the Lab - Vector Filler Sequence

<400> 28

cttctgaggc ggaaagaacc agatcctctc ttaaggtagc atcgagattt aaattaggga 60

taacagggta atggcgcggg ccgc 84

<210> 29

<211> 63

<212> DNA

<213> Artificial sequence

<220>

<223> Made in the Lab - Vector Filler Sequence

<400> 29

gttacccagg ctggagtgca gtggcacatt tctgctcact gcaacctcct cctccctggg 60

ttc 63

<210> 30

<211> 573

<212> DNA

<213> Artificial sequence

<220>

<223> Lab Made – CAG Promoter in Human Betaherpesvirus 5

<400> 30

acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat 60

aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga 120

gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc 180

ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt 240

atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga 300

ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt 360

gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggcgcg 420

cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg 480

cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc 540

ggccctataa aaagcgaagc gcgcggcggg cgg 573

<210> 31

<211> 771

<212> DNA

<213> Artificial sequence

<220>

<223> Lab Made – MHCK7 Promoter

<400> 31

acccttcaga ttaaaaataa ctgaggtaag ggcctgggta ggggaggtgg tgtgagacgc 60

tcctgtctct cctctatctg cccatcggcc ctttggggag gaggaatgtg cccaaggact 120

aaaaaaaggc catggagcca gaggggcgag ggcaacagac ctttcatggg caaaccttgg 180

ggccctgctg tctagcatgc cccactacgg gtctaggctg cccatgtaag gaggcaaggc 240

ctggggacac ccgagatgcc tggttataat taacccagac atgtggctgc cccccccccc 300

ccaacacctg ctgcctctaa aaataaccct gtccctggtg gatcccctgc atgcgaagat 360

cttcgaacaa ggctgtgggg gactgagggc aggctgtaac aggcttgggg gccagggctt 420

atacgtgcct gggactccca aagtattact gttccatgtt cccggcgaag ggccagctgt 480

cccccgccag ctagactcag cacttagttt aggaaccagt gagcaagtca gcccttgggg 540

cagcccatac aaggccatgg ggctgggcaa gctgcacgcc tgggtccggg gtgggcacgg 600

tgcccgggca acgagctgaa agctcatctg ctctcagggg cccctccctg gggacagccc 660

ctcctggcta gtcacaccct gtaggctcct ctatataacc caggggcaca ggggctgccc 720

tcattctacc accacctcca cagcacagac agacactcag gagccagcca g 771

<210> 32

<211> 544

<212> DNA

<213> Homo sapiens

<400> 32

ctcagtccat taggagccag tagcctggaa gatgtcttta cccccagcat cagttcaagt 60

ggagcagcac ataactcttg ccctctgcct tccaagattc tggtgctgag acttatggag 120

tgtcttggag gttgccttct gccccccaac cctgctccca gctggccctc ccaggcctgg 180

gttgctggcc tctgctttat caggattctc aagagggaca gctggtttat gttgcatgac 240

tgttccctgc atatctgctc tggttttaaa tagcttatct gagcagctgg aggaccacat 300

gggcttatat ggcgtggggt acatgttcct gtagccttgt ccctggcacc tgccaaaata 360

gcagccaaca ccccccaccc ccaccgccat ccccctgccc cacccgtccc ctgtcgcaca 420

ttcctccctc cgcagggctg gctcaccagg ccccagccca catgcctgct taaagccctc 480

tccatcctct gcctcaccca gtccccgctg agactgagca gacgcctcca ggatctgtcg 540

gcag 544

<210> 33

<211> 502

<212> DNA

<213> Homo sapiens

<400> 33

ctcagtccat taggagccag tagcctggaa gatgtcttta cccccagcat cagttcaagt 60

ggagcagcac ataactcttg ccctctgcct tccaagattc tggtgctgag acttatggag 120

tgtcttggag gttgccttct gccccccaac cctgctccca gctggccctc ccaggcctgg 180

gttgctggcc tctgctttat caggattctc aagagggaca gctggtttat gttgcatgac 240

tgttccctgc atatctgctc tggttttaaa tagcttatct gagcagctgg aggaccacat 300

gggcttatat ggcgtggggt acatgttcct gtagccttgt ccctggcacc tgccaaaata 360

gcagccaaca ccccccaccc ccaccgccat ccccctgccc cacccgtccc ctgtcgcaca 420

ttcctccctc cgcagggctg gctcaccagg ccccagccca catgcctgct taaagccctc 480

tccatcctct gcctcaccca gt 502

<210> 34

<211> 5464

<212> DNA

<213> Mus musculus

<400> 34

ggtaccggat cctgcaaggt cacacaaggg tctccaccca ccaggtgccc tagtctcaat 60

ttcagtttcc atgccttgtt ctcacaatgc tggcctcccc agagctaatt tggactttgt 120

ttttatttca aaagggcctg aatgaggagt agatcttgtg ctacccagct ctaagggtgc 180

ccgtgaagcc ctcagacctg gagcctttgc aacagccctt taggtggaag cagaataaag 240

caattttcct taaagccaaa atcctgcctc tagactcttc ttctctgacc tcggtccctg 300

ggctctaggg tggggaggtg gggcttggaa gaagaaggtg gggaagtggc aaaagccgat 360

ccctagggcc ctgtgaagtt cggagccttc cctgtacagc actggctcat agatcctcct 420

ccagccaaac atagcaagaa gtgatacctc ctttgtgact tccccaggcc cagtacctgt 480

caggttgaaa caggatttag agaagcctct gaactcacct gaactctgaa gctcatccac 540

caagcaagca cctaggtgcc actgctagtt agtatcctac gctgataata tgcagagctg 600

ggccacagaa gtcctggggt gtaggaactg accagtgact tttcagtcgg caaaggtatg 660

accccctcag cagatgtagt aatgtcccct tagatcccat cccaggcagg tctctaagag 720

gacatgggat gagagatgta gtcatgtggc attccaaaca cagctatcca cagtgtccct 780

tgccccttcc acttagccag gaggacagta accttagcct atctttcttc ctccccatcc 840

tccccaggaca caccccctgg tctgcagtat tcatttcttc cttcacgtcc cctctgtgac 900

ttccattgc aaggcttttg acctctgcag ctgctggaag atagagtttg gccctaggtg 960

tggcaagcca tctcaagaga aagcagacaa cagggggacc agattttgga aggatcagga 1020

actaaatcac tggcggggcct gggggtagaa aaaagagtga gtgagtccgc tccagctaag 1080

ccaagctagt ccccgagata ctctgccaca gctgggctgc tcggggtagc tttaggaatg 1140

tgggtctgaa agacaatggg attggaagac atctctttga gtctcccctc aaccccacct 1200

acagacacac tcgtgtgtgg ccagactcct gttcaacagc cctctgtgtt ctgaccactg 1260

agctaggcaa ccagagcatg ggccctgtgc tgaggatgaa gagttggtta ccaatagcaa 1320

aaacagcagg ggagggagaa cagagaacga aataaggaag gaagaaggaa aggccagtca 1380

atcagatgca gtcagaagag atgggaagcc aacacacagc ttgagcagag gaaacagaaa 1440

agggagagat tctgggcata aggaggccac agaaagaaga gcccaggccc cccaagtctc 1500

ctctttatac cctcatcccg tctcccaatt aagcccactc ttcttcctag atcagacctg 1560

agctgcagcg aagagacccg tagggaggat cacactggat gaaggagatg tgtggagaag 1620

tccagggaac ctaagagcca gagcctaaaa gagcaagaga taaaggtgct tcaaaggtgg 1680

ccaggctgtg cacacagagg gtcgaggact ggtggtagag cctcaagata aggatgatgc 1740

tcagaatggg cggggggggg gattctgggg gggggagaga gaaggtgaga aggagcctgg 1800

aacagagaat ctggaagcgc tggaaacgat accataaagg gaagaaccca ggctaccttt 1860

agatgtaaat catgaaagac agggagaagg gaagctggag agagtagaag gaccccgggg 1920

caagacattg aagcaaggac aagccaggtt gagcgctccg tgaaatcagc ctgctgaagg 1980

cagagccctg gtatgagcac cagaacagca gaggctaggg ttaatgtcga gacagggaac 2040

agaaggtaga cacaggaaca gacagagacg ggggagccag gtaacaaagg aatggtcctt 2100

ctcacctgtg gccagagcgt ccatctgtgt ccacatactc tagaatgttc atcagactgc 2160

agggctggct tgggaggcag ctggaaagag tatgtgagag ccaggggaga caagggggcc 2220

taggaaagga agaagagggc aaaccaggcc acacaagagg gcagagccca gaactgagtt 2280

aactccttcc ttgttgcatc ttccatagga ggcagtggga actctgtgac caccatcccc 2340

catgagcccc cactacccat accaagtttg gcctgagtgg cattctaggt tccctgagga 2400

cagagcctgg cctttgtctc ttggacctga cccaagctga cccaatgttc tcagtacctt 2460

atcatgccct caagagcttg agaaccaggc agtgacatat taggccatgg gctaaccctg 2520

gagcttgcac acaggagcct caagtgacct ccagggacac agctgcagac aggtggcctt 2580

tatccccaaa gagcaaccat ttggcatagg tggctgcaaa tgggaatgca aggttgaatc 2640

aggtcccttc aagaatactg catgcaagac ctaagacccc tggagagagg ggtatgctcc 2700

tgcccccacc caccataagg ggagtgaact atcctagggg gctggcgacc ttggggagac 2760

accacattac tgagagtgct gagcccagaa aaactgaccg ccctgtgtcc tgcccacctc 2820

cacactctag agctatattg agaggtgaca gtagataggg tgggagctgg tagcagggag 2880

agtgttcctg ggtgtgaggg tgtaggggaa agccagagca ggggagtctg gctttgtctc 2940

ctgaacacaa tgtctactta gttataacag gcatgacctg ctaaagaccc aacatctacg 3000

acctctgaaa agacagcagc cctggaggac aggggttgtc tctgagcctt gggtgcttga 3060

tggtgccaca aaggaggca tgagtgtgag tataaggccc caggagcgtt agagaagggc 3120

acttgggaag gggtcagtct gcagagcccc tatccatgga atctggagcc tggggccaac 3180

tggtgtaaat ctctgggcct gccaggcatt caaagcagca cctgcatcct ctggcagcct 3240

ggggaggcgg aagggagcaa ccccccactt ataccctttc tccctcagcc ccaggattaa 3300

cacctctggc cttccccctt cccacctccc atcaggagtg gagggttgca gagggagggt 3360

aaaaacctac atgtccaaac atcatggtgc acgatatatg gatcagtatg tgtagaggca 3420

agaaaggaaa tctgcaggct taactgggtt aatgtgtaaa gtctgtgtgc atgtgtgtgt 3480

gtctgactga aaacgggcat ggctgtgcag ctgttcagtt ctgtgcgtga ggttaccaga 3540

ctgcaggttt gtgtgtaaat tgcccaaggc aaagtgggtg aatcccttcc atggtttaaa 3600

gagatggat gatggcctgc atctcaagga ccatggaaaa tagaatggac actctatatg 3660

tgtctctaag ctaaggtagc aaggtctttg gaggacacct gtctagagat gtgggcaaca 3720

gagactacag acagtatctg tacagagtaa ggagagagag gagggggtgt agaattctct 3780

tactatcaaa gggaaactga gtcgtgcacc tgcaaagtgg atgctctccc tagacatcat 3840

gactttgtct ctggggagcc agcactgtgg aacttcaggt ctgagagagt aggaggctcc 3900

cctcagcctg aagctatgca gatagccagg gttgaaaggg ggaagggaga gcctgggatg 3960

ggagcttgtg tgttggaggc aggggacaga tattaagcct ggaagagaag gtgaccctta 4020

cccagttgtt caactcaccc ttcagattaa aaataactga ggtaagggcc tgggtagggg 4080

aggtggtgtg agacgctcct gtctctcctc tatctgccca tcggcccttt ggggaggagg 4140

aatgtgccca aggactaaaa aaaggccatg gagccagagg ggcgagggca acagaccttt 4200

catgggcaaa ccttggggcc ctgctgtcct cctgtcacct cgagccaa gggatcaaag 4260

gaggagggagc caggacagga gggaagtggg agggagggtc ccagcagagg actccaaatt 4320

taggcagcag gcatatggga tgggatataa aggggctgga gcactgagag ctgtcagaga 4380

tttctccaac ccaggtaaga gggagtttcg ggtggggggct cttcacccac accagacctc 4440

tccccaccta gaaggaaact gcctttcctg gaagtggggt tcaggccggt cagagatctg 4500

acagggtggc cttccaccag cctgggaagt tctcagtggc aggaggtttc cacaagaaac 4560

actggatgcc ccttccctta cgctgtcttc tccatcttcc tcctggggat gctcctcccc 4620

gtcttggttt atcttggctc ttcgtcttca gcaagatttg ccctgtgctg tccactccat 4680

ctttctctac tgtctccgtg ccttgccttg ccttcttgcg tgtccttcct ttccacccat 4740

ttctcacttc accttttctc cccttctcat ttgtattcat ccttccttcc ttccttcctt 4800

ccttccttcc ttccttcctt ccttcctttc tcccttcctt ccttccttcc ttccttcctt 4860

ccttccttcc ttcctgtgtc agagtgctga gaatcacacc tggggttccc acccttatgt 4920

aaacaatctt ccagtgagcc acagcttcag tgctgctggg tgctctctta ccttcctcac 4980

cccctggctt gtcctgttcc atcctggtca ggatctctag attggtctcc cagcctctgc 5040

tactcctctt cctgcctgtt cctctctctg tccagctgcg ccactgtggt gcctcgttcc 5100

agctgtggtc cacattcttc aggattctct gaaaagttaa ccaggtgaga atgtttcccc 5160

tgtagacagc agatcacgat tctcccggaa gtcaggcttc cagccctctc tttctctgcc 5220

cagctgcccg gcactcttag caaacctcag gcacccttac cccacataga cctctgacag 5280

agaagcaggc actttacatg gagtcctggt gggagcca taggctacgg tgtaaaagag 5340

gcagggaagt ggtggtgtag gaaagtcagg acttcacata gaagcctagc ccacaccaga 5400

aatgacagac agatccctcc tatctccccc ataagagttt gagtcgaccc gcggccccga 5460

attg 5464

<210> 35

<211> 413

<212> DNA

<213> Gallus gallus

<400> 35

gggataaaag cagtctgggc tttcacatga cagcatctgg ggctgcggca gagggtcggg 60

tccgaagcgc tgccttatca gcgtccccag ccctgggagg tgacagctgg ctggcttgtg 120

tcagcccctc gggcactcac gtatctccgt ccgacgggtt taaaatagca aaactctgag 180

gccacacaat agcttgggct tatatgggct cctgtggggg aagggggagc acggaggggg 240

ccggggccgc tgctgccaaa atagcagctc acaagtgttg cattcctctc tgggcgccgg 300

gcacattcct gctggctctg cccgccccgg ggtgggcgcc ggggggacct taaagcctct 360

gccccccaag gagcccttcc cagacagccg ccggcaccca ccgctccgtg gga 413

<210> 36

<211> 1090

<212> DNA

<213> Homo sapiens

<400> 36

ctctcagccc tggaagtcct tgctcacagc cgaggcgccg agagcgcttg ctctgcccag 60

atctgcgcga gtctggcgcc cgcgctctga acggcgtcgc tgcccagccc ccttccccgg 120

gaggtgggag cggccaccca gggccccgtg gctgcccttg taaggaggcg aggcccgagg 180

acacccgaga cgcccggtta taattaacca ggacacgtgg cgaacccccc tccaacacct 240

gcccccgaac ccccccatac ccagcgcctc gggtctcggc ctttgcggca gaggagacag 300

caaagcgccc tctaaaaata actcctttcc cggcgaccga gaccctccct gtcccccgca 360

cagcggaaat ctcccagtgg caccgagggg gcgagggtta agtgggggggg agggtgacca 420

ccgcctccca cccttgccct gagtttgaat ctctccaact cagccagcct cagtttcccc 480

tccactcagt ccctaggagg aaggggcgcc caagcgcggg tttctggggt tagactgccc 540

tccattgcaa ttggtccttc tcccggcctc tgcttcctcc agctcacagg gtatctgctc 600

ctcctggagc cacaccttgg ttccccgagg tgccgctggg actcgggtag gggtgagggc 660

ccaggggggca cagggggagc cgaggggccac aggaagggct ggtggctgaa ggagactcag 720

gggccagggg acggtggctt ctacgtgctt gggacgttcc cagccaccgt cccatgttcc 780

cggcgggggg ccagctgtcc ccaccgccag cccaactcag cacttggtca gggtatcagc 840

ttggtggggg ggcgtgagcc cagcccctgg ggcggctcag cccatacaag gccatggggc 900

tgggcgcaaa gcatgcctgg gttcagggtg ggtatggtgc gggagcaggg aggtgagagg 960

ctcagctgcc ctccagaact cctccctggg gacaacccct cccagccaat agcacagcct 1020

aggtccccct atataaggcc acggctgctg gcccttcctt tgggtcagtg tcacctccag 1080

gatacagaca 1090

<210> 37

<211> 253

<212> DNA

<213> Homo sapiens

<400> 37

gcccagcacc ccaaggcggc caacgccaaa actctccctc ctcctcttcc tcaatctcgc 60

tctcgctctt tttttttttc gcaaaaggag gggagagggg gtaaaaaaat gctgcactgt 120

gcggcgaagc cggtgagtga gcggcgcggg gccaatcagc gtgcgccgtt ccgaaagttg 180

ccttttatgg ctcgagcggc cgcggcggcg ccctataaaa cccagcggcg cgacgcgcca 240

ccaccgccga gtc 253

<210> 38

<211> 281

<212> DNA

<213> Gallus gallus

<400> 38

ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc 60

aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg 120

ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg 180

cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc 240

ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg a 281

<210> 39

<211> 220

<212> DNA

<213> Human betaherpesvirus 5

<400> 39

tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat 60

ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 120

actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac 180

ggtgggaggt ctatataagc agagctcgtt tagtgaaccg 220

<210> 40

<211> 583

<212> DNA

<213> Human betaherpesvirus 5

<400> 40

tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 60

cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 120

gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 180

atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 240

aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 300

catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 360

catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 420

atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 480

ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 540

acggtgggag gtctatataa gcagagctgg tttagtgaac cgt 583

<210> 41

<211> 508

<212> DNA

<213> Human betaherpesvirus 5

<400> 41

cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 60

gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 120

atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 180

aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 240

catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 300

catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 360

atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 420

ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 480

acggtgggag gtctatataa gcagagct 508

<210> 42

<211> 573

<212> DNA

<213> Artificial sequence

<220>

<223> Lab Made – CAG Promoter in Human Betaherpesvirus 5

<400> 42

acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat 60

aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga 120

gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc 180

ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt 240

atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga 300

ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt 360

gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggcgcg 420

cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg 480

cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc 540

ggccctataa aaagcgaagc gcgcggcggg cgg 573

<210> 43

<211> 580

<212> DNA

<213> Artificial sequence

<220>

<223> Lab Made – CAG Promoter in Human Betaherpesvirus 5

<400> 43

cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 60

gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 120

atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 180

aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 240

catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 300

catgtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc 360

ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg 420

ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg 480

tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg 540

gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg 580

<210> 44

<211> 455

<212> DNA

<213> Homo sapiens

<400> 44

caacctttgg agctaagcca gcaatggtag agggaagatt ctgcacgtcc cttccaggcg 60

gcctccccgt caccaccccc cccaacccgc cccgaccgga gctgagagta attcatacaa 120

aaggactcgc ccctgccttg gggaatccca gggaccgtcg ttaaactccc actaacgtag 180

aacccagaga tcgctgcgtt cccgccccct cacccgcccg ctctcgtcat cactgaggtg 240

gagaatagca tgcgtgaggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag 300

tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga aggtggcgcg 360

gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag 420

aaccgtatat aagtgcagta gtcgccgtga acgtt 455

<210> 45

<211> 401

<212> DNA

<213> Homo sapiens

<400> 45

agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga 60

ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga 120

gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg 180

gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc 240

gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg 300

cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg 360

cgaccatctg cgctgcggcg ccggcgactc agcgctgcct c 401

<210> 46

<211> 448

<212> DNA

<213> Homo sapiens

<400> 46

agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga 60

ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga 120

gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg 180

gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc 240

gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg 300

cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg 360

cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg 420

aggagtcgtg tcgtgcctga gagcgcag 448

<210> 47

<211> 422

<212> DNA

<213> Homo sapiens

<400> 47

ctgcagaggg ccctgcgtat gagtgcaagt gggttttagg accaggatga ggcggggtgg 60

gggtgcctac ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca 120

aattgcgcat cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc 180

actgccagct tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg 240

ccgcctcagc actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc 300

cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg 360

cgagataggg gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc 420

tc 422

<210> 48

<211> 281

<212> DNA

<213> Homo sapiens

<400> 48

acttgtggac aaagtttgct ctattccacc tcctccaggc cctccttggg tccatcaccc 60

caggggtgct gggtccatcc cacccccagg cccacacagg cttgcagtat tgtgtgcggt 120

atggtcaggg cgtccgagag caggtttcgc agtggaaggc aggcaggtgt tggggaggca 180

gttaccgggg caacgggaac agggcgtttt ggaggtggtt gccatgggga cctggatgct 240

gacgaaggct cgcgaggctg tgagcagcca cagtgccctg c 281

<210> 49

<211> 851

<212> DNA

<213> Artificial sequence

<220>

<223> Lab-Made – eSYN Promoter Polynucleotide

<400> 49

gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 60

catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 120

acgacccccg cccatgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 180

ctttccatg acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc 240

aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 300

ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 360

tagtcatcgc tattaccatg gctgcagagg gccctgcgta tgagtgcaag tgggttttag 420

gaccaggatg aggcggggtg ggggtgccta cctgacgacc gaccccgacc cactggacaa 480

gcacccaacc cccattcccc aaattgcgca tcccctatca gagaggggga ggggaaacag 540

gatgcggcga ggcgcgtcgc gactgccagc ttcagcaccg cggacagtgc cttcgccccc 600

gcctggcggc gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt cactcgccgg 660

tcccccgcaa actcccttc ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc 720

cggaccgcac cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg 780

cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggagagtcg tgtcgtgcct 840

gagagcgcag g 851

<210> 50

<211> 304

<212> DNA

<213> Human betaherpesvirus 5

<400> 50

cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 60

gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 120

atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 180

aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 240

catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 300

catg 304

<210> 51

<211> 953

<212> DNA

<213> Homo sapiens

<400> 51

cgcgtccgcc cgcgagcaca gagcctcgcc tttgccgatc cgccgcccgt ccacacccgc 60

cgccaggtaa gcccggccag ccgaccgggg catgcggccg cggcccttcg cccgtgcaga 120

gccgccgtct gggccgcagc ggggggcgca tggggcggaa ccggaccgcc gtggggggcg 180

cggggagaagc ccctgggcct ccggagatgg gggacacccc acgccagttc gcaggcgcga 240

ggccgcgctc gggcgggcgc gctccggggg tgccgctctc ggggcggggg caaccggcgg 300

ggtctttgtc tgagccgggc tcttgccaat ggggatcgca cggtgggcgc ggcgtagccc 360

ccgtcaggcc cggtgggggc tggggcgcca tgcgcgtgcg cgctggtcct ttgggcgcta 420

actgcgtgcg cgctgggaat tggcgctaat tgcgcgtgcg cgctgggact caatggcgct 480

aatcgcgcgt gcgttctggg gcccgggcgc ttgcgccact tcctgcccga gccgctggcg 540

cccgaggtg tggccgctgc gtgcgcgcgc gcgacccggt cgctgtttga accgggcgga 600

ggcggggctg gcgcccggtt gggagggggt tggggcctgg cttcctgccg cgcgccgcgg 660

ggacgcctcc gaccagtgtt tgccttttat ggtaataacg cggccggccc ggcttccttt 720

gtccccaatc tgggcgcgcg ccggcgcccc ctggcggcct aaggactcgg cgcgccggaa 780

gtggccaggg cggcagcggc tgctcttggc ggccccgagg tgactatagc cttcttttgt 840

gtcttgatag ttcgccagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc 900

tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttc 953

<210> 52

<211> 1068

<212> DNA

<213> Artificial sequence

<220>

<223> Lab Made - Chicken Beta Actin Exon/Intron Plus Intron

rabbit globin

<400> 52

gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc 60

cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg 120

ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc 180

cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg 240

tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc 300

gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc 360

ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc 420

gtggggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc 480

cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc 540

gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg 600

ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc 660

tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag 720

ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc 780

tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc 840

cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg 900

ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc 960

ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg 1020

ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattc 1068

<210> 53

<211> 149

<212> DNA

<213> Artificial sequence

<220>

<223> Lab Made - Chimeric Intron Sequence

<400> 53

ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg gtgcaaatca 60

aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa gtgttacttc 120

tgctctaaaa gctgcggaat tgtacccgc 149

<210> 54

<211> 126

<212> DNA

<213> Homo sapiens

<400> 54

agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagctg tgctcctggg 60

caccgcgcag tccgcccccg cggctcctgg ccagaccacc cctaggaccc cctgccccaa 120

gtcgca 126

<210> 55

<211> 121

<212> DNA

<213> Human betaherpesvirus 5

<400> 55

tcagatcgcc tggagaggcc atccacgctg ttttgacctc catagtggac accgggaccg 60

atccagcctc cgcggccggg aacggtgcat tggaacgcgg attccccgtg ccaagagtga 120

c 121

<210> 56

<211> 512

<212> DNA

<213> Artificial sequence

<220>

<223> Lab-Made - Adenovirus-Derived Enhancer Element

<400> 56

ctcactctct tccgcatcgc tgtctgcgag ggccagctgt tgggctcgcg gttgaggaca 60

aactcttcgc ggtctttcca gtactcttgg atcggaaacc cgtcggcctc cgaacggtac 120

tccgccaccg agggacctga gcgagtccgc atcgaccgga tcggaaaacc tctcgagaaa 180

ggcgtctaac cagtcacagt cgcaaggtag gctgagcacc gtggcgggcg gcagcgggtg 240

gcggtcgggg ttgtttctgg cggaggtgct gctgatgatg taattaaagt aggcggtctt 300

gagacggcgg atggtcgagg tgaggtgtgg caggcttgag atccagctgt tggggtgagt 360

actccctctc aaaagcgggc attacttctg cgctaagatt gtcagtttcc aaaaacgagg 420

aggatttgat attcacctgg cccgatctgg ccatacactt gagtgacaat gacatccact 480

ttgcctttct ctccacaggt gtccactccc ag 512

<210> 57

<211> 956

<212> DNA

<213> Homo sapiens

<400> 57

ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg 60

cctggcctct ttacggggtta tggcccttgc gtgccttgaa ttacttccac ctggctccag 120

tacgtgattc ttgatcccga gctggagcca ggggcggggcc ttgcgcttta ggagcccctt 180

cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt 240

ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat 300

gacgtgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caggatctgc 360

acactggtat ttcggttttt gggcccgcgg ccggcgacgg ggcccgtgcg tcccagcgca 420

catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc 480

aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg 540

cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc 600

ctgctccagg gggctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac 660

ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt 720

accgggcgcc gtccaggcac ctcgattagt tctggagctt ttggagtacg tcgtctttag 780

gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag 840

ttaggccagc ttggcacttg atgtaattct ccttggaatt tggccttttt gagtttggat 900

cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcag 956

<210> 58

<211> 939

<212> DNA

<213> Homo sapiens

<400> 58

gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg 60

ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga 120

agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt 180

gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt 240

ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt 300

tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt 360

tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg 420

ggcctgcgag cgcggccacc gagaatcgga cggggggtagt ctcaagctgg ccggcctgct 480

ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg 540

tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca 600

aaatggagga cgcggcgctc ggggagagcgg gcgggtgagt cacccacaca aaggaaaagg 660

gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg 720

cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt 780

tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac 840

ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag 900

cctcagacag tggttcaaag tttttttctt ccatttcag 939

<210> 59

<211> 83

<212> DNA

<213> Homo sapiens

<400> 59

tcagaagccc cgggctcgtc agtcaaaccg gttctctgtt tgcactcggc agcacgggca 60

ggcaagtggt ccctaggttc ggg 83

<210> 60

<211> 476

<212> DNA

<213> Homo sapiens

<400> 60

gtgagtctat gggacccttg atgttttctt tccccttctt ttctatggtt aagttcatgt 60

cataggaagg ggagaagtaa cagggtacac atattgacca aatcagggta attttgcatt 120

tgtaatttta aaaaatgctt tcttctttta atatactttt ttgtttatct tatttctaat 180

actttcccta atctctttct ttcagggcaa taatgataca atgtatcatg cctctttgca 240

ccattctaaa gaataacagt gataatttct gggttaaggc aatagcaata tttctgcata 300

taaatatttc tgcatataaa ttgtaactga tgtaagaggt ttcatattgc taatagcagc 360

tacaatccag ctaccattct gcttttattt tatggttggg ataaggctgg attattctga 420

gtccaagcta ggcccttttg ctaatcatgt tcatacctct tatcttcctc ccacag 476

<210> 61

<211> 196

<212> DNA

<213> monkey virus 40

<400> 61

tctagaggat ccggtactcg aggaactgaa aaaccagaaa gttaactggt aagtttagtc 60

tttttgtctt ttatttcagg tcccggatcc ggtggtggtg caaatcaaag aactgctcct 120

cagtggatgt tgcctttact tctaggcctg tacggaagtg ttacttctgc tctaaaagct 180

gcggaattgt acccgc 196

<210> 62

<211> 589

<212> DNA

<213> Artificial sequence

<220>

<223> Lab-Made - Mutated Regulatory Element of Woodchuck Hepatitis

<400> 62

aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 60

ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 120

atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 180

tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 240

ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 300

attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 360

ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc 420

gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 480

aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 540

cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgc 589

<210> 63

<211> 588

<212> DNA

<213> Artificial sequence

<220>

<223> Lab-Made - Mutated Regulatory Element of Woodchuck Hepatitis

<400> 63

tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 60

ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 120

ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 180

gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 240

ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 300

tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacaggg ctcggctgtt 360

gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 420

ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 480

tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 540

ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgca 588

<210> 64

<211> 755

<212> DNA

<213> Artificial sequence

<220>

<223> Lab-Made - Mutated Regulatory Element of Woodchuck Hepatitis

<400> 64

ttcctgttaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 60

atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 120

cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 180

aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 240

cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 300

ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacaggg 360

ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccgc 420

ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 480

cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 540

cgcctcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgccca 600

tgtatctttt tcacctgtgc cttgtttttg cctgtgttcc gcgtcctact tttcaagcct 660

ccaagctgtg ccttgggcgg ctttggggca tggacataga tccctataaa gaatttggtt 720

catcttatca gttgttgaat tttcttcctt tggac 755

<210> 65

<211> 12

<212> DNA

<213> Artificial sequence

<220>

<223> CAAX Motive

<400> 65

tgtgtgataa tg 12

<210> 66

<211> 810

<212> DNA

<213> Homo sapiens

<400> 66

ctgttctcat cacatcatat caaggttata taccatcaat attgccacag atgttactta 60

gccttttaat atttctctaa tttagtgtat atgcaatgat agttctctga tttctgagat 120

tgagtttctc atgtgtaatg attatttaga gtttctcttt catctgttca aatttttgtc 180

tagttttatt ttttactgat ttgtaagact tctttttata atctgcatat tacaattctc 240

tttactgggg tgttgcaaat attttctgtc attctatggc ctgacttttc ttaatggttt 300

tttaatttta aaaataagtc ttaatattca tgcaatctaa ttaacaatct tttctttgtg 360

gttaggactt tgagtcataa gaaatttttc tctacactga agtcatgatg gcatgcttct 420

atattatttt ctaaaagatt taaagttttg ccttctccat ttagacttat aattcactgg 480

aatttttttg tgtgtatggt atgacatatg ggttcccttt tattttttac atataaatat 540

atttccctgt ttttctaaaa aagaaaaaga tcatcatttt cccattgtaa aatgccatat 600

ttttttcata ggtcacttac atatatcaat gggtctgttt ctgagctcta ctctatttta 660

tcagcctcac tgtctatccc cacacatctc atgctttgct ctaaatcttg atatttagtg 720

gaacattctt tcccattttg ttctacaaga atatttttgt tattgtcttt gggctttcta 780

tatacatttt gaaatgaggt tgacaagtta 810

<210> 67

<211> 726

<212> DNA

<213> Hepatitis B virus

<400> 67

ataacaggcc tattgattgg aaagtttgtc aacgaattgt gggtcttttg gggtttgctg 60

ccccttttac gcaatgtgga tatcctgctt taatgccttt atatgcatgt atacaagcaa 120

aacaggcttt tactttctcg ccaacttaca aggcctttct cagtaaacag tatatgaccc 180

tttaccccgt tgctcggcaa cggcctggtc tgtgccaagt gtttgctgac gcaaccccca 240

ctggttgggg cttggccata ggccatcagc gcatgcgtgg aacctttgtg tctcctctgc 300

cgatccatac tgcggaactc ctagccgctt gttttgctcg cagcaggtct ggagcaaacc 360

tcatcgggac cgacaattct gtcgtactct cccgcaagta tacatcgttt ccatggctgc 420

taggctgtgc tgccaactgg atcctgcgcg ggacgtcctt tgtttacgtc ccgtcggcgc 480

tgaatcccgc ggacgacccc tcccggggcc gcttggggct ctaccgcccg cttctccgtc 540

tgccgtaccg tccgaccacg gggcgcacct ctctttacgc ggactccccg tctgtgcctt 600

ctcatctgcc ggaccgtgtg cacttcgctt cacctctgca cgtcgcatgg aggccaccgt 660

gaacgcccac cggaacctgc ccaaggtctt gcataagagg actcttggac tttcagcaat 720

gtcatc 726

<210> 68

<211> 755

<212> DNA

<213> Artificial sequence

<220>

<223> Lab-Made - Enhancer Element Derived from HepB

<400> 68

ttcctgtaaa caggcctatt gattggaaag tttgtcaacg aattgtgggt cttttggggt 60

ttgctgcccc ttttacgcaa tgtggatatc ctgctttaat gcctttatat gcatgtatac 120

aagcaaaaca ggcttttact ttctcgccaa cttacaaggc ctttctcagt aaacagtata 180

tgacccttta ccccgttgct cggcaacggc ctggtctgtg ccaagtgttt gctgacgcaa 240

cccccactgg ttggggcttg gccataggcc atcagcgcat gcgtggaacc tttgtgtctc 300

ctctgccgat ccatactgcg gaactcctag ccgcttgttt tgctcgcagc tggactggag 360

caaacctcat cgggaccgac aattctgtcg tactctcccg caagcactca ccgtttccgc 420

ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 480

cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 540

cgcctcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgccca 600

tgtatctttt tcacctgtgc cttgtttttg cctgtgttcc gcgtcctact tttcaagcct 660

ccaagctgtg ccttgggcgg ctttggggca tggacataga tccctataaa gaatttggtt 720

catcttatca gttgttgaat tttcttcctt tggac 755

<210> 69

<211> 94

<212> DNA

<213> Homo sapiens

<400> 69

gctggagcct cggtagccgt tcctcctgcc cgctgggcct cccaacgggc cctcctcccc 60

tccttgcacc ggcccttcct ggtctttgaa taaa 94

<210> 70

<211> 596

<212> DNA

<213> Marmot hepatitis virus

<400> 70

attcgagcat cttaccgcca tttattccca tatttgttct gtttttcttg atttgggtat 60

acatttaaat gttaataaaa caaaatggtg gggcaatcat ttacattttt agggatatgt 120

aattactagt tcaggtgtat tgccacaaga caaacatgtt aagaaacttt cccgttattt 180

acgctctgtt cctgttaatc aacctctgga ttacaaaatt tgtgaaagat tgactgatat 240

tcttaactat gttgctcctt ttacgctgtg tggatatgct gctttaatgc ctctgtatca 300

tgctattgct tcccgtacgg ctttcgtttt ctcctccttg tataaatcct ggttgctgtc 360

tctttatgag gagttgtggc ccgttgtccg tcaacgtggc gtggtgtgct ctgtgtttgc 420

tgacgcaacc cccactggct ggggcattgc caccacctgt caactccttt ctgggacttt 480

cgctttcccc ctcccgatcg ccacggcaga actcatcgcc gcctgccttg cccgctgctg 540

gacaggggct aggttgctgg gcactgataa ttccgtggtg ttgtcgggga agggcc 596

<210> 71

<211> 387

<212> DNA

<213> Oryctolagus cuniculus

<400> 71

tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc 60

actcggaaga acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta 120

gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg 180

tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac 240

ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa 300

attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat 360

agctgtccct cttctcttat ggagatc 387

<210> 72

<211> 251

<212> DNA

<213> Bos taurus

<400> 72

ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac 60

tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca 120

ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aatacaatag 180

caggcatgct ggggatgcgg tgggctctat gggtacccag gtgctgaaga attgacccgg 240

ttcctcctgg g 251

<210> 73

<211> 251

<212> DNA

<213> Bos taurus

<400> 73

ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac 60

tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca 120

ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag 180

caggcatgct ggggatgcgg tgggctctat gggtacccag gtgctgaaga attgacccgg 240

ttcctcctgg g 251

<210> 74

<211> 225

<212> DNA

<213> Bos taurus

<400> 74

ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc 60

tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc 120

tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt 180

gggaagacaa tagcaggcat gctggggatg cggtgggctc tatgg 225

<210> 75

<211> 202

<212> DNA

<213> Homo sapiens

<400> 75

ctgcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc 60

actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag 120

gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcccaagttg 180

ggaagaaacc tgtagggcct gc 202

<210> 76

<211> 735

<212> PROTEIN

<213> Adeno-associated virus 2

<400> 76

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Thr Leu Ser

1 5 10 15

Glu Gly Ile Arg Gln Trp Trp Lys Leu Lys Pro Gly Pro Pro Pro Pro

20 25 30

Lys Pro Ala Glu Arg His Lys Asp Asp Ser Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Glu Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Arg Gln Leu Asp Ser Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Pro Val Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu His Ser Pro Val Glu Pro Asp Ser Ser Ser Gly Thr Gly

145 150 155 160

Lys Ala Gly Gln Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Ala Asp Ser Val Pro Asp Pro Gln Pro Leu Gly Gln Pro Pro

180 185 190

Ala Ala Pro Ser Gly Leu Gly Thr Asn Thr Met Ala Thr Gly Ser Gly

195 200 205

Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Thr Trp Met Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Ser Gln Ser Gly Ala Ser Asn Asp Asn His Tyr

260 265 270

Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe His

275 280 285

Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn Trp

290 295 300

Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile Gln Val

305 310 315 320

Lys Glu Val Thr Gln Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn Leu

325 330 335

Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro Tyr

340 345 350

Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala Asp

355 360 365

Val Phe Met Val Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly Ser

370 375 380

Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro Ser

385 390 395 400

Gln Met Leu Arg Thr Gly Asn Asn Phe Thr Phe Ser Tyr Thr Phe Glu

405 410 415

Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp Arg

420 425 430

Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg Thr

435 440 445

Asn Thr Pro Ser Gly Thr Thr Thr Gln Ser Arg Leu Gln Phe Ser Gln

450 455 460

Ala Gly Ala Ser Asp Ile Arg Asp Gln Ser Arg Asn Trp Leu Pro Gly

465 470 475 480

Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Ser Ala Asp Asn Asn

485 490 495

Asn Ser Glu Tyr Ser Trp Thr Gly Ala Thr Lys Tyr His Leu Asn Gly

500 505 510

Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Ser His Lys Asp

515 520 525

Asp Glu Glu Lys Phe Phe Pro Gln Ser Gly Val Leu Ile Phe Gly Lys

530 535 540

Gln Gly Ser Glu Lys Thr Asn Val Asp Ile Glu Lys Val Met Ile Thr

545 550 555 560

Asp Glu Glu Glu Ile Arg Thr Thr Asn Pro Val Ala Thr Glu Gln Tyr

565 570 575

Gly Ser Val Ser Thr Asn Leu Gln Arg Gly Asn Arg Gln Ala Ala Thr

580 585 590

Ala Asp Val Asn Thr Gln Gly Val Leu Pro Gly Met Val Trp Gln Asp

595 600 605

Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His Thr

610 615 620

Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu Lys

625 630 635 640

His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala Asn

645 650 655

Pro Ser Thr Thr Phe Ser Ala Ala Lys Phe Ala Ser Phe Ile Thr Gln

660 665 670

Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln Lys

675 680 685

Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn Tyr

690 695 700

Asn Lys Ser Val Asn Val Asp Phe Thr Val Asp Thr Asn Gly Val Tyr

705 710 715 720

Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 77

<211> 736

<212> PROTEIN

<213> Adeno-associated virus 9

<400> 77

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Gly Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln

580 585 590

Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn

690 695 700

Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val

705 710 715 720

Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

725 730 735

<210> 78

<211> 736

<212> PROTEIN

<213> Adeno-associated virus 6

<400> 78

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro

20 25 30

Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro

115 120 125

Phe Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Ile Gly

145 150 155 160

Lys Thr Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro

180 185 190

Ala Thr Pro Ala Ala Val Gly Pro Thr Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ala

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Ser Ala Ser Thr Gly Ala Ser Asn Asp Asn His

260 265 270

Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe

275 280 285

His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn

290 295 300

Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile Gln

305 310 315 320

Val Lys Glu Val Thr Thr Asn Asp Gly Val Thr Thr Ile Ala Asn Asn

325 330 335

Leu Thr Ser Thr Val Gln Val Phe Ser Asp Ser Glu Tyr Gln Leu Pro

340 345 350

Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala

355 360 365

Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly

370 375 380

Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro

385 390 395 400

Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Thr Phe Ser Tyr Thr Phe

405 410 415

Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp

420 425 430

Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Asn Arg

435 440 445

Thr Gln Asn Gln Ser Gly Ser Ala Gln Asn Lys Asp Leu Leu Phe Ser

450 455 460

Arg Gly Ser Pro Ala Gly Met Ser Val Gln Pro Lys Asn Trp Leu Pro

465 470 475 480

Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Lys Thr Asp Asn

485 490 495

Asn Asn Ser Asn Phe Thr Trp Thr Gly Ala Ser Lys Tyr Asn Leu Asn

500 505 510

Gly Arg Glu Ser Ile Ile Asn Pro Gly Thr Ala Met Ala Ser His Lys

515 520 525

Asp Asp Lys Asp Lys Phe Phe Pro Met Ser Gly Val Met Ile Phe Gly

530 535 540

Lys Glu Ser Ala Gly Ala Ser Asn Thr Ala Leu Asp Asn Val Met Ile

545 550 555 560

Thr Asp Glu Glu Glu Ile Lys Ala Thr Asn Pro Val Ala Thr Glu Arg

565 570 575

Phe Gly Thr Val Ala Val Asn Leu Gln Ser Ser Ser Thr Asp Pro Ala

580 585 590

Thr Gly Asp Val His Val Met Gly Ala Leu Pro Gly Met Val Trp Gln

595 600 605

Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His

610 615 620

Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu

625 630 635 640

Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala

645 650 655

Asn Pro Pro Ala Glu Phe Ser Ala Thr Lys Phe Ala Ser Phe Ile Thr

660 665 670

Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln

675 680 685

Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Val Gln Tyr Thr Ser Asn

690 695 700

Tyr Ala Lys Ser Ala Asn Val Asp Phe Thr Val Asp Asn Asn Gly Leu

705 710 715 720

Tyr Thr Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Pro Leu

725 730 735

<210> 79

<211> 738

<212> PROTEIN

<213> Non-human primate adeno-associated virus

<400> 79

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro

20 25 30

Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Pro Ser Pro Gln Arg Ser Pro Asp Ser Ser Thr Gly Ile

145 150 155 160

Gly Lys Lys Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln

165 170 175

Thr Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro

180 185 190

Pro Ala Gly Pro Ser Gly Leu Gly Ser Gly Thr Met Ala Ala Gly Gly

195 200 205

Gly Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser

210 215 220

Ser Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val

225 230 235 240

Ile Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His

245 250 255

Leu Tyr Lys Gln Ile Ser Asn Gly Thr Ser Gly Gly Ser Thr Asn Asp

260 265 270

Asn Thr Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn

275 280 285

Arg Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn

290 295 300

Asn Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn

305 310 315 320

Ile Gln Val Lys Glu Val Thr Gln Asn Glu Gly Thr Lys Thr Ile Ala

325 330 335

Asn Asn Leu Thr Ser Thr Ile Gln Val Phe Thr Asp Ser Glu Tyr Gln

340 345 350

Leu Pro Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe

355 360 365

Pro Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn

370 375 380

Asn Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr

385 390 395 400

Phe Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr

405 410 415

Gln Phe Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser

420 425 430

Leu Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu

435 440 445

Ser Arg Thr Gln Ser Thr Gly Gly Thr Ala Gly Thr Gln Gln Leu Leu

450 455 460

Phe Ser Gln Ala Gly Pro Asn Asn Met Ser Ala Gln Ala Lys Asn Trp

465 470 475 480

Leu Pro Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Thr Thr Leu Ser

485 490 495

Gln Asn Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His

500 505 510

Leu Asn Gly Arg Asp Ser Leu Val Asn Pro Gly Val Ala Met Ala Thr

515 520 525

His Lys Asp Asp Glu Glu Arg Phe Phe Pro Ser Ser Gly Val Leu Met

530 535 540

Phe Gly Lys Gln Gly Ala Gly Lys Asp Asn Val Asp Tyr Ser Ser Val

545 550 555 560

Met Leu Thr Ser Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr

565 570 575

Glu Gln Tyr Gly Val Val Ala Asp Asn Leu Gln Gln Gln Asn Ala Ala

580 585 590

Pro Ile Val Gly Ala Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val

595 600 605

Trp Gln Asn Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile

610 615 620

Pro His Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe

625 630 635 640

Gly Leu Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val

645 650 655

Pro Ala Asp Pro Pro Thr Thr Phe Ser Gln Ala Lys Leu Ala Ser Phe

660 665 670

Ile Thr Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu

675 680 685

Leu Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr

690 695 700

Ser Asn Tyr Tyr Lys Ser Thr Asn Val Asp Phe Ala Val Asn Thr Asp

705 710 715 720

Gly Thr Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg

725 730 735

Asn Leu

<210> 80

<211> 2217

<212> DNA

<213> Non-human primate adeno-associated virus

<400> 80

atggctgccg atggttatct tccagattgg ctcgaggaca acctctctga gggcattcgc 60

gagtggtggg acctgaaacc tggagccccg aaacccaaag ccaaccagca aaagcaggac 120

aacggccggg gtctggtgct tcctggctac aagtacctcg gacccttcaa cggactcgac 180

aagggggagc ccgtcaacgc ggcggacgca gcggccctcg agcacgacaa ggcctacgac 240

cagcagctcc aagcgggtga caatccgtac ctgcggtata atcacgccga cgccgagttt 300

caggagcgtc tgcaagaaga tacgtctttt gggggcaacc tcgggcgcgc agtcttccag 360

gccaaaaagc gggttctcga acctctgggc ctggttgaat cgccggttaa gacggctcct 420

ggaaagaaga gaccggtaga gccatcaccc cagcgctctc cagactcctc tacgggcatc 480

ggcaagaaag gccagcagcc cgcaaaaaag agactcaatt ttgggcagac tggcgactca 540

gagtcagtcc ccgaccctca accaatcgga gaaccaccag caggcccctc tggtctggga 600

tctggtacaa tggctgcagg cggtggcgct ccaatggcag acaataacga aggcgccgac 660

ggagtgggta gttcctcagg aaattggcat tgcgattcca catggctggg cgacagagtc 720

atcaccacca gcacccgcac ctgggccctg cccacctaca acaaccacct ctacaagcaa 780

atctccaacg ggacctcggg aggaagcacc aacgacaaca cctacttcgg ctacagcacc 840

ccctggggggt attttgactt caacagattc cactgccact tttcaccacg tgactggcag 900

cgactcatca acaacaactg gggattccgg cccaagaggc tcaacttcaa gctcttcaac 960

atccaagtca aggaggtcac gcagaatgaa ggcaccaaga ccatcgccaa taaccttacc 1020

agcacgattc aggtctttac ggactcggaa taccagctcc cgtacgtgct cggctcggcg 1080

caccagggct gcctgcctcc gttcccggcg gacgtcttca tgattcctca gtacgggtac 1140

ctgactctga acaatggcag tcaggctgtg ggccggtcgt ccttctactg cctggagtac 1200

tttccttctc aaatgctgag aacgggcaac aactttgaat tcagctacaa cttcgaggac 1260

gtgcccttcc acagcagcta cgcgcacagc cagagcctgg accggctgat gaaccctctc 1320

atcgaccagt acttgtacta cctgtcccgg actcaaagca cgggcggtac tgcaggaact 1380

cagcagttgc tattttctca ggccggggcct aacaacatgt cggctcaggc caagaactgg 1440

ctacccggtc cctgctaccg gcagcaacgc gtctccacga cactgtcgca gaacaacaac 1500

agcaactttg cctggacggg tgccaccaag tatcatctga atggcagaga ctctctggtg 1560

aatcctggcg ttgccatggc tacccacaag gacgacgaag agcgattttt tccatccagc 1620

ggagtcttaa tgtttgggaa acagggagct ggaaaagaca acgtggacta tagcagcgtg 1680

atgctaacca gcgaggaaga aataaagacc accaacccag tggccacaga acagtacggc 1740

gtggtggccg ataacctgca acagcaaaac gccgctccta ttgtaggggc cgtcaatagt 1800

caagggagcct tacctggcat ggtgtggcag aaccgggacg tgtacctgca gggtcccatc 1860

tgggccaaga ttcctcatac ggacggcaac tttcatccct cgccgctgat gggaggcttt 1920

ggactgaagc atccgcctcc tcagatcctg attaaaaaca cacctgttcc cgcggatcct 1980

ccgaccacct tcaatcaggc caagctggct tctttcatca cgcagtacag taccggccag 2040

gtcagcgtgg agatcgagtg ggagctgcag aaggagaaca gcaaacgctg gaacccagag 2100

attcagtaca cttccaacta ctacaaatct acaaatgtgg actttgctgt caatactgag 2160

ggtacttatt ccgagcctcg ccccattggc acccgttacc tcacccgtaa tctgtaa 2217

<210> 81

<211> 535

<212> PROTEIN

<213> Non-human primate adeno-associated virus

<400> 81

Met Ala Ala Gly Gly Gly Ala Pro Met Ala Asp Asn Asn Glu Gly Ala

1 5 10 15

Asp Gly Val Gly Ser Ser Ser Gly Asn Trp His Cys Asp Ser Thr Trp

20 25 30

Leu Gly Asp Arg Val Ile Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro

35 40 45

Thr Tyr Asn Asn His Leu Tyr Lys Gln Ile Ser Asn Gly Thr Ser Gly

50 55 60

Gly Ser Thr Asn Asp Asn Thr Tyr Phe Gly Tyr Ser Thr Pro Trp Gly

65 70 75 80

Tyr Phe Asp Phe Asn Arg Phe His Cys His Phe Ser Pro Arg Asp Trp

85 90 95

Gln Arg Leu Ile Asn Asn Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn

100 105 110

Phe Lys Leu Phe Asn Ile Gln Val Lys Glu Val Thr Gln Asn Glu Gly

115 120 125

Thr Lys Thr Ile Ala Asn Asn Leu Thr Ser Thr Ile Gln Val Phe Thr

130 135 140

Asp Ser Glu Tyr Gln Leu Pro Tyr Val Leu Gly Ser Ala His Gln Gly

145 150 155 160

Cys Leu Pro Pro Phe Pro Ala Asp Val Phe Met Ile Pro Gln Tyr Gly

165 170 175

Tyr Leu Thr Leu Asn Asn Gly Ser Gln Ala Val Gly Arg Ser Ser Phe

180 185 190

Tyr Cys Leu Glu Tyr Phe Pro Ser Gln Met Leu Arg Thr Gly Asn Asn

195 200 205

Phe Glu Phe Ser Tyr Asn Phe Glu Asp Val Pro Phe His Ser Ser Tyr

210 215 220

Ala His Ser Gln Ser Leu Asp Arg Leu Met Asn Pro Leu Ile Asp Gln

225 230 235 240

Tyr Leu Tyr Tyr Leu Ser Arg Thr Gln Ser Thr Gly Gly Thr Ala Gly

245 250 255

Thr Gln Gln Leu Leu Phe Ser Gln Ala Gly Pro Asn Asn Met Ser Ala

260 265 270

Gln Ala Lys Asn Trp Leu Pro Gly Pro Cys Tyr Arg Gln Gln Arg Val

275 280 285

Ser Thr Thr Leu Ser Gln Asn Asn Asn Ser Asn Phe Ala Trp Thr Gly

290 295 300

Ala Thr Lys Tyr His Leu Asn Gly Arg Asp Ser Leu Val Asn Pro Gly

305 310 315 320

Val Ala Met Ala Thr His Lys Asp Asp Glu Glu Arg Phe Phe Pro Ser

325 330 335

Ser Gly Val Leu Met Phe Gly Lys Gln Gly Ala Gly Lys Asp Asn Val

340 345 350

Asp Tyr Ser Ser Val Met Leu Thr Ser Glu Glu Glu Ile Lys Thr Thr

355 360 365

Asn Pro Val Ala Thr Glu Gln Tyr Gly Val Val Ala Asp Asn Leu Gln

370 375 380

Gln Gln Asn Ala Ala Pro Ile Val Gly Ala Val Asn Ser Gln Gly Ala

385 390 395 400

Leu Pro Gly Met Val Trp Gln Asn Arg Asp Val Tyr Leu Gln Gly Pro

405 410 415

Ile Trp Ala Lys Ile Pro His Thr Asp Gly Asn Phe His Pro Ser Pro

420 425 430

Leu Met Gly Gly Phe Gly Leu Lys His Pro Pro Pro Gln Ile Leu Ile

435 440 445

Lys Asn Thr Pro Val Pro Ala Asp Pro Pro Thr Thr Phe Asn Gln Ala

450 455 460

Lys Leu Ala Ser Phe Ile Thr Gln Tyr Ser Thr Gly Gln Val Ser Val

465 470 475 480

Glu Ile Glu Trp Glu Leu Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro

485 490 495

Glu Ile Gln Tyr Thr Ser Asn Tyr Tyr Lys Ser Thr Asn Val Asp Phe

500 505 510

Ala Val Asn Thr Glu Gly Thr Tyr Ser Glu Pro Arg Pro Ile Gly Thr

515 520 525

Arg Tyr Leu Thr Arg Asn Leu

530 535

<210> 82

<211> 398

<212> PROTEIN

<213> Non-human primate adeno-associated virus

<400> 82

Ser Thr Ile Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro Tyr Val

1 5 10 15

Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala Asp Val

20 25 30

Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly Ser Gln

35 40 45

Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro Ser Gln

50 55 60

Met Leu Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr Asn Phe Glu Asp

65 70 75 80

Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp Arg Leu

85 90 95

Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg Thr Gln

100 105 110

Ser Thr Gly Gly Thr Ala Gly Thr Gln Gln Leu Leu Phe Ser Gln Ala

115 120 125

Gly Pro Asn Asn Met Ser Ala Gln Ala Lys Asn Trp Leu Pro Gly Pro

130 135 140

Cys Tyr Arg Gln Gln Arg Val Ser Thr Thr Leu Ser Gln Asn Asn Asn

145 150 155 160

Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His Leu Asn Gly Arg

165 170 175

Asp Ser Leu Val Asn Pro Gly Val Ala Met Ala Thr His Lys Asp Asp

180 185 190

Glu Glu Arg Phe Phe Pro Ser Ser Gly Val Leu Met Phe Gly Lys Gln

195 200 205

Gly Ala Gly Lys Asp Asn Val Asp Tyr Ser Ser Val Met Leu Thr Ser

210 215 220

Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Gln Tyr Gly

225 230 235 240

Val Val Ala Asp Asn Leu Gln Gln Gln Asn Ala Ala Pro Ile Val Gly

245 250 255

Ala Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val Trp Gln Asn Arg

260 265 270

Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His Thr Asp

275 280 285

Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu Lys His

290 295 300

Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala Asp Pro

305 310 315 320

Pro Thr Thr Phe Asn Gln Ala Lys Leu Ala Ser Phe Ile Thr Gln Tyr

325 330 335

Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln Lys Glu

340 345 350

Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn Tyr Tyr

355 360 365

Lys Ser Thr Asn Val Asp Phe Ala Val Asn Thr Glu Gly Thr Tyr Ser

370 375 380

Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

385 390 395

<210> 83

<211> 332

<212> PROTEIN

<213> Non-human primate adeno-associated virus

<400> 83

Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr Asn Phe Glu Asp Val Pro

1 5 10 15

Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp Arg Leu Met Asn

20 25 30

Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg Thr Gln Ser Thr

35 40 45

Gly Gly Thr Ala Gly Thr Gln Gln Leu Leu Phe Ser Gln Ala Gly Pro

50 55 60

Asn Asn Met Ser Ala Gln Ala Lys Asn Trp Leu Pro Gly Pro Cys Tyr

65 70 75 80

Arg Gln Gln Arg Val Ser Thr Thr Leu Ser Gln Asn Asn Asn Ser Asn

85 90 95

Phe Ala Trp Thr Gly Ala Thr Lys Tyr His Leu Asn Gly Arg Asp Ser

100 105 110

Leu Val Asn Pro Gly Val Ala Met Ala Thr His Lys Asp Asp Glu Glu

115 120 125

Arg Phe Phe Pro Ser Ser Gly Val Leu Met Phe Gly Lys Gln Gly Ala

130 135 140

Gly Lys Asp Asn Val Asp Tyr Ser Ser Val Met Leu Thr Ser Glu Glu

145 150 155 160

Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Gln Tyr Gly Val Val

165 170 175

Ala Asp Asn Leu Gln Gln Gln Asn Ala Ala Pro Ile Val Gly Ala Val

180 185 190

Asn Ser Gln Gly Ala Leu Pro Gly Met Val Trp Gln Asn Arg Asp Val

195 200 205

Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His Thr Asp Gly Asn

210 215 220

Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu Lys His Pro Pro

225 230 235 240

Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala Asp Pro Pro Thr

245 250 255

Thr Phe Asn Gln Ala Lys Leu Ala Ser Phe Ile Thr Gln Tyr Ser Thr

260 265 270

Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln Lys Glu Asn Ser

275 280 285

Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn Tyr Tyr Lys Ser

290 295 300

Thr Asn Val Asp Phe Ala Val Asn Thr Glu Gly Thr Tyr Ser Glu Pro

305 310 315 320

Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu

325 330

<210> 84

<211> 743

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic construct - AAV9 variant

<400> 84

Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser

1 5 10 15

Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro

20 25 30

Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro

35 40 45

Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro

50 55 60

Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp

65 70 75 80

Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala

85 90 95

Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly

100 105 110

Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro

115 120 125

Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg

130 135 140

Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly

145 150 155 160

Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr

165 170 175

Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro

180 185 190

Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly

195 200 205

Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser

210 215 220

Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile

225 230 235 240

Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu

245 250 255

Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn

260 265 270

Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg

275 280 285

Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn

290 295 300

Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile

305 310 315 320

Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn

325 330 335

Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu

340 345 350

Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro

355 360 365

Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp

370 375 380

Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe

385 390 395 400

Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu

405 410 415

Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu

420 425 430

Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser

435 440 445

Arg Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser

450 455 460

Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro

465 470 475 480

Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn

485 490 495

Asn Asn Ser Glu Phe Ala Trp Pro Gly Ala Ser Ser Trp Ala Leu Asn

500 505 510

Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys

515 520 525

Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly

530 535 540

Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile

545 550 555 560

Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser

565 570 575

Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Thr Leu Ala Val

580 585 590

Pro Phe Lys Ala Gln Ala Gln Thr Gly Trp Val Gln Asn Gln Gly Ile

595 600 605

Leu Pro Gly Met Val Trp Gln Asp Arg Asp Val Tyr Leu Gln Gly Pro

610 615 620

Ile Trp Ala Lys Ile Pro His Thr Asp Gly Asn Phe His Pro Ser Pro

625 630 635 640

Leu Met Gly Gly Phe Gly Met Lys His Pro Pro Pro Gln Ile Leu Ile

645 650 655

Lys Asn Thr Pro Val Pro Ala Asp Pro Pro Thr Ala Phe Asn Lys Asp

660 665 670

Lys Leu Asn Ser Phe Ile Thr Gln Tyr Ser Thr Gly Gln Val Ser Val

675 680 685

Glu Ile Glu Trp Glu Leu Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro

690 695 700

Glu Ile Gln Tyr Thr Ser Asn Tyr Tyr Lys Ser Asn Asn Val Glu Phe

705 710 715 720

Ala Val Asn Thr Glu Gly Val Tyr Ser Glu Pro Arg Pro Ile Gly Thr

725 730 735

Arg Tyr Leu Thr Arg Asn Leu

740

<210> 85

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Peptide insert

<400> 85

Thr Leu Ala Val Pro Phe Lys

1 5

<210> 86

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Peptide insert

<400> 86

Lys Phe Pro Val Ala Leu Thr

1 5

<---

Claims (70)

1. A polynucleotide for increasing the expression of muscle protein LIM (MLP), comprising an expression cassette, wherein the polynucleotide comprises a polynucleotide sequence encoding MLP, operably linked to a promoter. 2. The polynucleotide of claim 1, wherein the promoter is a heart-specific promoter. 3. The polynucleotide of claim 1 or 2, wherein the promoter is a muscle-specific promoter. 4. The polynucleotide of any one of claims 1 to 3, wherein the promoter is a cardiomyocyte-specific promoter. 5. The polynucleotide of any one of claims 1 to 4, wherein the promoter is the MHCK7 promoter. 6. The polynucleotide of claim 5, wherein the MHCK7 promoter has at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 31. 7. The polynucleotide of any one of claims 1-4, wherein the promoter is the cardiac troponin T (hTNNT2) promoter. 8. The polynucleotide of claim 7, wherein the hTNNT2 promoter has at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 32. 9. The polynucleotide of any one of claims 1-4, 7 or 8, wherein the expression cassette comprises exon 1 of the cardiac troponin T (hTNNT2) gene, wherein the hTNNT2 promoter and exon 1 together have at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 32. 10. The polynucleotide of any one of claims 1-4, wherein the promoter is a CMV promoter or a CAG promoter. 11. The polynucleotide of any one of claims 1-10, wherein the expression cassette comprises a polyA signal. 12. The polynucleotide of claim 11, wherein the polyA signal is human growth hormone (hGH) polyA. 13. The polynucleotide of any one of claims 1-12, wherein the expression cassette comprises a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE). 14. The polynucleotide of claim 1, wherein the MLP is a human MLP. 15. The polynucleotide of claim 14, wherein MLP is isoform A of MLP. 16. The polynucleotide of claim 14 or 15, wherein the MLP is characterized by at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 1. 17. The polynucleotide of claim 14 or 16, wherein the MLP is isoform B of MLP. 18. The polynucleotide of claim 15 or 17, wherein the MLP is characterized by at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 2. 19. The polynucleotide of claim 14, wherein the MLP is characterized by at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 3. 20. The polynucleotide of claim 14, wherein the MLP is characterized by at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 4. 21. The polynucleotide of any one of claims 1 to 20, wherein the polynucleotide sequence encoding the MLP is a cysteine-glycine-rich protein 3 (CSRP3) polynucleotide. 22. The polynucleotide of claim 21, wherein the CSRP3 polynucleotide is a human CSRP3 polynucleotide. 23. The polynucleotide of any one of claims 1-22, wherein the polynucleotide sequence encoding the MLP is characterized by at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 5. 24. The polynucleotide of any one of claims 1 to 23, wherein the polynucleotide sequence encoding the MLP is characterized by at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 7. 25. The polynucleotide of any one of claims 1 to 24, wherein the polynucleotide comprises at least about 2.4 kb, no more than about 2.6 kb, or from about 2.4 kb to about 2.6 kb. 26. The polynucleotide of any one of claims 1 to 25, wherein the polynucleotide comprises at least about 3.0 kb, no more than about 3.3 kb, or from about 3.0 kb to about 3.3 kb. 27. The polynucleotide of any one of claims 1 to 26, wherein the polynucleotide comprises at least about 2.4 kb, at least about 2.6 kb, at least about 3.0 kb, at least about 3.3 kb, at least about 3.5 kb, at least about 3.7 kb, at least about 3.9 kb, at least about 4.1 kb, or at least about 4.3 kb. 28. The polynucleotide of any one of claims 1 to 27, wherein the polynucleotide comprises at least about 2.6 kb, at least about 3.0 kb, no more than about 3.3 kb, no more than about 3.5 kb, no more than about 3.7 kb, no more than about 3.9 kb, no more than about 4.1 kb, no more than about 4.3 kb, or no more than about 4.5 kb. 29. The polynucleotide of any one of claims 1-28, wherein the expression cassette has at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identity to any one of SEQ ID NOs: 8-11. 30. The polynucleotide of any one of claims 1-29, wherein the polynucleotide has at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identity to any one of SEQ ID NOs: 12-15. 31. The polynucleotide of any one of claims 1-30, wherein the expression cassette is flanked by 5′ and 3′ AAV2 inverted terminal repeats (ITRs), wherein the ITRs have at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identity to any one of SEQ ID NOs: 20-26. 32. A polynucleotide according to any one of claims 1 to 31, wherein the polynucleotide is self-complementary. 33. The polynucleotide of any one of claims 1 to 32, wherein the polynucleotide comprises an expression cassette and a sequence reverse complementary to the expression cassette. 34. The polynucleotide of claim 33, wherein the expression cassette and the sequence reverse complementary to the expression cassette are flanked by 5′ and 3′ AAV2 inverted terminal repeats (ITRs), wherein the ITRs are characterized by at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 23 or SEQ ID NO: 26. 35. A gene therapy vector for increasing MLP expression, comprising a polynucleotide according to any one of claims 1-34. 36. The vector of claim 35, wherein the gene therapy vector is a recombinant adeno-associated virus (rAAV) vector. 37. The vector of claim 36, wherein the rAAV-based vector is AAV9. 38. The vector of claim 37, wherein the rAAV-based vector comprises a capsid protein that has 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to any of SEQ ID NO: 77. 39. The vector of claim 36, wherein the rAAV-based vector is an AAVrh10 vector. 40. The vector of claim 39, wherein the rAAV-based vector comprises a capsid protein that has 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to any of SEQ ID NO: 79. 41. The vector of claim 36, wherein the rAAV-based vector is an AAV6 vector. 42. The vector of claim 41, wherein the rAAV-based vector comprises a capsid protein that has 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to any of SEQ ID NO: 78. 43. The vector of claim 36, wherein the rAAV-based vector is an AAVrh74 vector. 44. The vector of claim 43, wherein the rAAV-based vector comprises a capsid protein that has 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to any of SEQ ID NO: 80. 45. The vector of any one of claims 36 to 44, wherein the rAAV-based vector is a self-complementary AAV-based vector. 46. A method of treating or preventing a disease or disorder in a subject having or suspected of having a disorder associated with defects in CSRP3, comprising administering to the subject a vector according to any one of claims 35-45. 47. The method of claim 46, wherein the disease or disorder is a cardiac disorder. 48. The method of claim 46 or 47, wherein the disease or disorder is heart failure. 49. The method of any one of claims 46-48, wherein the disease or disorder is hypertrophic cardiomyopathy. 50. The method according to any one of claims 46-48, wherein the disease or disorder is dilated cardiomyopathy. 51. The method according to any one of paragraphs 46-50, wherein the subject is a mammal. 52. The method according to claim 51, wherein the subject is a primate. 53. The method according to paragraph 52, where the subject is a human being. 54. The method according to any one of claims 46-53, wherein the subject has a mutation in the CSRP3 gene that causes an amino acid substitution selected from C58G, L44P, S54R, E55G and/or K69R, relative to human CSRP3 encoding a human MLP having the sequence of SEQ ID NO: 1. 55. The method according to any one of claims 46-54, wherein the vector is administered by intravenous injection, intracardiac injection, intracardiac infusion and/or cardiac catheterization. 56. The method of any one of claims 46-55, wherein the administration increases MLP expression by at least about 5%. 57. The method of any one of claims 46-56, wherein the administration increases MLP expression by at least about 30%. 58. The method of any one of claims 46-57, wherein the administration increases MLP expression by at least about 70%. 59. The method of any one of claims 46-58, wherein the administration increases MLP expression by about 5% to about 10%. 60. The method of any one of claims 46-57, wherein the administration increases MLP expression by about 30% to about 50%. 61. The method of any one of claims 46-58, wherein the administration increases MLP expression by about 70% to about 100%. 62. The method according to any one of claims 46-61, wherein the method provides for the treatment and/or prevention of a disease or disorder. 63. A pharmaceutical composition for increasing the expression of MLP, comprising a vector according to any one of claims 35–45. 64. A kit for increasing MLP expression, containing a vector according to any one of claims 35–45 or a pharmaceutical composition according to claim 63 and instructions for use. 65. The use of a composition according to claim 63 in the treatment of a disease or disorder by the method according to any of claims 46-62. 66. A method for expressing muscle LIM protein (MLP), comprising contacting a cell with a vector according to any one of claims 35-45. 67. The method according to claim 66, wherein the cell is a cardiomyocyte. 68. The method according to claim 67, wherein the cardiomyocyte is a human cardiomyocyte. 69. The method according to any one of claims 66-68, wherein the promoter is the MHCK7 promoter and wherein the level of expression of MLP is at least 2-fold higher than the level of expression of MLP in a cell transduced with a vector with the hTNNT2 promoter. 70. The method according to any one of claims 66-68, wherein the promoter is the MHCK7 promoter and wherein the level of expression of MLP is 2-10 times higher than the level of expression of MLP in a cell transduced with a vector with the hTNNT2 promoter.