CN110662835A - 工程化改造用于由增强的指导RNA优化的基因编辑和转录调节的最小化SaCas9 CRISPR/Cas系统 - Google Patents

Abstract

提供了通过利用一组紧凑Cas9衍生物对Cas9功能的可编程和精确调节，所述紧凑Cas9衍生物通过基于跨多种2类CRISPR效应子的结构信息来使保守的HNH和/或REC‑C结构域缺失而产生。还提供了通过分割的mini‑dSaCas9并且紧接分割点之后融合FokI结构域来工程化改造二聚gRNA指导的核酸酶的新策略，以便由于二聚FokI核酸酶与靶序列接近度更近而提高目标DNA切割效率并潜在地降低脱靶效应。通过将优化的紧凑gRNA表达盒与尺寸缩小的SaCas9衍生物组合，将具有用于反式激活、DNA切割和碱基编辑的不同效应子结构域的整个CRISPR/Cas系统装载到单个AAV病毒中。这样的仅用一个AAV‑CRISPR/Cas9系统在需要安全有效的体内递送的生物医学应用中特别有吸引力。

Description

工程化改造用于由增强的指导RNA优化的基因编辑和转录调 节的最小化SaCas9 CRISPR/Cas系统

技术领域

本申请涉及生物学领域，特别涉及遗传物质的定向修饰。更特别地，要求保护的发明涉及对Cas9 CRISPR相关蛋白及其相关产物的改进。

背景技术

从化脓性链球菌(Streptococcus pyogenes)中发现的CRISPR相关蛋白9(Cas9)是一种多结构域蛋白质，由于其卓越的模块化和多功能性，其已广泛用于哺乳动物细胞的基因组编辑和转录控制。由于尺寸限制，体内递送合成的基因线路受到限制，特别是对于有效负载能力几乎等于整个Cas9复合体的较小的递送系统。

发明内容

技术问题

已经开发了多种策略以通过调节dCas9和gRNA表达来工程化改造哺乳动物细胞中的模块化和分层基因线路。哺乳动物细胞中的转录控制可通过将转录调节结构域与核酸酶失活的 Cas9(dCas9)直接融合来实现。或者，可以通过用重复肽支架标记dCas9或通过将重复RNA 基序与同源gRNA融合来将多个转录调节结构域募集到dCas9。然而，CRISPR/Cas系统的生物医学应用需要探索新的平台以用于工程化改造整合并处理多个内源输入的哺乳动物合成线路。另外，由于现有病毒递送载体的限制性货物尺寸，CRISPR/Cas治疗线路的应用也具有挑战性。

问题的解决方案

分割的Cas9系统通常可用于绕过病毒递送载体的包装限制，并且在要求保护的发明中，将dCas9分割并且在人细胞中重构。治疗应用的挑战之一是找到可以将所有CRISPR/Cas9 组分携带到期望的器官或细胞群体以进行基因操纵的最佳递送系统。当合并至病毒递送系统中时，以最大潜力使用CRISPR/Cas系统受到其物理尺寸的极大限制。当在具有位点特异性整合的高价值递送系统(例如腺相关病毒/AAV)中用于合成生物学目的时，整个cas9复合体类似于计算机操纵系统，其占用了95％的可用内存，仅留下一小部分用于合成生物学编程目的。通过将CRISPR/CAS9分割成多个较小的区域并将各区域递送在分开的病毒传递载体中，这保留了强大的基因操纵功能，同时大幅增加了用于细胞编程目的的空间。所要求保护的发明显示了对现有CAS9递送技术的实质性改进，并且包括了对基因控制和编程的其它增强。

尽管已经采用了多种病毒递送系统并且具有不同的成功，但是依赖于替代病毒系统的系统的实施可能导致不希望的强烈免疫应答。使用重组腺相关病毒(rAAV)具有很高的基因转移效率和非常低的免疫应答。不幸的是，包装能力被限制在4.7kb至5kb，相对于超过4.2kb的人类优化Cas9尺寸并且加上启动子序列达到超过5kb的限制，这是存在问题的。使用内含肽介导的分割Cas9，内含肽起蛋白质内含子的作用并且被从序列中切除并通过肽键连接其余的侧翼区(外显肽)而不会留下疤痕。关于分割位点的选择，由于蛋白质剪接的空间需要，需特别留意表面暴露的分割位点。这种系统允许Cas9的编码序列分布在双载体或多载体系统上并且在翻译后重构。

所要求保护的发明经由基因线路的改进和更高水平的基因线路递送增强通过靶向特异性诊断和治疗应用而扩大了合成生物学的可及之处。本发明要求保护的实施例通过Cas9的最佳分割克服了现有的尺寸限制，从而允许由较小的递送系统容纳更高水平的合成基因线路。

本要求保护的发明利用了通过缺失保守功能结构域而保留了DNA结合活性的尺寸缩小的金黄色葡萄球菌Cas9变体(mini-SaCas9)。在一个优选的说明性实施例中，将FokI核酸酶结构域融合至分割mini-SaCas9的中间以触发有效的DNA切割。在另一个说明性实施例中，基因编辑系统足够小以被容纳在包含与尺寸缩小的反式激活结构域融合的mini-SaCas9 以及具有有效反式激活活性的优化的紧凑gRNA表达盒的单个AAV中。所要求保护的发明突出了一种产生用于体内应用的具有不同效应子结构域的仅用一个AAV-CRISPR/Cas9系统的实用方法。

为了绕过AAV有效负载限制，将来自化脓性链球菌(Streptococcus pyogenes)的4.2kb Cas9(SpCas9)分割并与指导RNA(gRNA)表达单元一起包装到两个单独的AAV中，其允许在体内对全长SpCas9进行功能重构。另一种策略是寻找尺寸减小的天然2类CRISPR 效应子，例如通过使用宏基因组数据集在未经培养的生物体中鉴定出的3.2-kb SaCas9和～3-kb CasX。为了进一步减小转基因尺寸，可以使用～70-bp谷氨酰胺tRNA代替～250bp的 RNA聚合酶III启动子来驱动tRNA:gRNA融合转录物的表达，所述tRNA:gRNA融合转录物被内源tRNase Z切割以产生活性gRNA。这些努力促进了用于CRISPR/Cas技术的体内应用的仅用一个AAV递送载体的构建。

对SpCas9、SaCas9和氨基酸球菌属(Acidaminococcus sp.)Cpf1(AsCpf1)的最近结构研究阐明了这些2类CRISPR效应子中保守结构域(包括分别切割互补DNA链和非互补DNA链的HNH/NUC和RuvC核酸酶结构域)、识别(REC)结构域和前间隔序列邻近基序 (PAM)相互作用(PI)结构域的功能。有趣的是，通过缺失HNH或REC2结构域的截短型SpCas9突变体基本上保留了完整的DNA结合活性或一半切割活性。这些结果突出了将野生型Cas9进一步尺寸缩小至仅具有DNA结合活性而没有DNA切割活性的最小化Cas9 (mini-Cas9)的可能性，从而允许在单个AAV载体中容纳其它DNA模板、效应子结构域和控制元件。

发明的有益效果

在优选具有限制性货物尺寸的病毒递送载体的生物医学应用中，这种CRISPR/Cas9系统具有特别的实用性。预见到的变体包括分割的Cas9/dCas9系统与rAAV递送系统的组合， Cas9/dCas9的活性可以被控制以在体内编辑和调节内源基因。

附图说明

包括附图以更好地说明所要求保护的发明的说明性实施例。

图1是与VPR结构域融合的紧凑Cas9衍生物的EBFP2转录激活测定的图。

图2是dSpCas9、mini-dSpCas9-1和mini-dSpCas9-2结构域组织以及相应的基因激活效率的图。

图3是dSaCas9及其衍生物的结构域组织以及所得基因激活效率的示意图。

图4是dAsCpf1和mini-AsCpf1-1结构域组织以及相应的基因激活效率的图。

图5是优化的gRNA和相应的基因激活效率的示意图。

图6是谷氨酰胺(Glu)tRNA用作表达优化的gRNA-2的启动子的图。

图7是包括与FokI结构域融合的分割的dSaCas9和分割的mini-dSaCas9-4的dSaCas9 变体的结构域组织的图。

图8是证明DNA切割效率的EYFP重构测定的图以及与分割的dSaCas9和分割的minidSaCas9-4融合的二聚FokI核酸酶对DNA切割的图示。

图9是间隔区长度在12-bp至24-bp范围内的分割的dSaCas9变体的DNA切割效率的图。

图10是VPR、VTR1、VTR2和VTR3转录激活结构域以及使用EBFP2报告系统评估的其相应基因激活效率的示意图。

图11是用于转录激活的SpyTag和MoonTag重复阵列以及相应基因激活效率数据的示意图。

图12是通过使用装载有紧凑CRISPR/Cas9系统的单个AAV的转录激活的示意图以及 EBFP2激活效率的图示。

图13是将mini-Cas9、效应子结构域、gRNA表达盒和其它部分包装在单个AAV载体中以用于转录激活、DNA切割和碱基编辑的示意图。

图14是miniSaCas9-3和分割的SaCas9以及相应miniSaCas9-3和分割SaCas9激活效率的示意图。

图15是示出间隔区的CCR5基因的基因序列图示以及相应的凝胶琼脂糖图数据。

具体实施方式

如在相应附图中进一步详述的以下实施例中，进一步详细说明了紧凑CRISPR/Cas9系统的合理设计。

图1是与VPR结构域融合的紧凑Cas9衍生物的EBFP2转录激活测定的图(101)。将组成型表达的mKate2用作转染对照。在第一个说明性实例中，将报告系统用在培养的人胚肾293(HEK293)细胞中。通过分别缺失对于核酸酶失活Cas9(dCas9)的DNA结合活性非必要的REC1结构域C末端区域(REC-C,△501-710)和HNH结构域(△777-891)产生了两种mini-dSpCas9，并且这两种mini-dSpCas9分别与VP64-p65-Rta(VPR)反式激活结构域融合。

图2是dSpCas9、mini-dSpCas9-1和mini-dSpCas9-2结构域组织的图(201)以及相应基因激活效率的图示(203)。在该说明性实例中，与dSpCas9:VPR相比，这两种 mini-dSpCas9:VPR变体保留了超过50％的反式激活能力。

图3是dSaCas9及其衍生物的结构域组织的示意图(301)和描述基因激活效率的图示 (303)。近来，野生型SaCas9已经被工程化改造以识别改变的PAM(“NNNRRT”)。基于SaCas9突变体，公开的实施例通过用“GGGGSGGGG”接头(GS-接头)替换保守的REC-C 结构域(△234-444)构建了mini-SaCas9-1:VPR，其仅保留了dSaCas9:VPR的～2.5％的反式激活活性。尽管GS接头被广泛用作柔性接头，但其仍可能会使SaCas9结构变形。受最近称为SEWING的计算方案的启发，使用邻近残基搜索(ARS)方案来搜索不连续Cas9片段之间的现有结构。通过使用ARS方案将GS接头替换为适合于填充在REC-C缺失间隙中的来自SaCas9 BH结构域的“KRRRRHR”(R接头)，导致mini-SaCas9-2的反式激活能力提高至11倍。作为所要求保护的发明的增强的实施例，通过使用SAR方案，使用来源于风产液菌(Aquifex aeolicus)中推定基因(登录号，O67859)的“GSK接头”，其适合于HNH结构域的缺失间隙(△479-649)。通过缺失REC-C结构域和HNH结构域两者产生了2-kb mini-SaCas9-3，其相对于对照仅保留了0.6％的反式激活活性。

图4是dAsCpf1和mini-AsCpf1-1结构域组织的图(401)以及相应基因激活效率的图示(403)。为了在该说明性实施例中评估mini-SaCas9变体的DNA切割效率，使用了报告重构测定，其中DNA切割可以触发从非活性形式重构活性增强的黄色荧光蛋白(EYFP)报告基因。缺失REC-C或缺失REC-C结构域和HNH结构域两者会导致背景EYFP表达，表明结构域缺失消除了mini-SaCas9变体的DNA切割活性。在该说明性实施例中，通过缺失 REC2结构域(△324-525)，dAsCpf1:VPR的反式激活能力保留了46％，表明该缺失策略适用于独特的2类CRISPR效应子。

图5是优化的gRNA的示意图(501)以及基因激活效率的相应图示(503)。可以引入A-U翻转或U-G转换以破坏gRNA支架的第一个茎环中推定的RNA Pol III终止子序列，从而增强dCas9介导的DNA标记的效率。推定的RNA Pol III终止子序列显示在方框中，在第三位和第四位用阴影标记A-U和G-C突变。还在右侧示出了流式细胞术数据的代表性散点图。将组成型表达的mKate2用作转染对照。在一个优选的说明性实施例中，通过在推定的 RNAPol III终止子序列中引入一个或两个点突变，miniSaCas9-3的反式激活效率提高至约100倍。

图6是谷氨酰胺(Glu)tRNA(601)用作表达优化的gRNA-2的启动子的图。在该说明性实例中，将HEK293细胞用表达mini-SaCas9-3:VPR和被Glu RNA或U6启动子驱动的 gRNA-2的质粒共转染。右侧示出了代表性的流式细胞术散点图(603)。将组成型表达的iRFP 用作转染对照。数据显示为在转染入HEK293细胞后48小时使用流式细胞仪测量的三次独立重复的EBFP2荧光的平均值±SEM倍数变化。在该说明性实施例中，当使用谷氨酰胺tRNA 代替U6启动子来驱动gRNA表达时，获得～50％的反式激活活性。

改善CRISPR/Cas9系统的DNA切割特异性对于未来的临床应用至关重要。其中FokI核酸酶结构域与dSpCas9的N-末端融合而不与C-末端融合的杂合蛋白的二聚化改善了PAM外方向(PAM-out orientation)上的DNA切割特异性。此外，具有较短靶互补性区域的截短gRNA降低了脱靶切割效率。

在以下说明性实施例中，进一步公开了紧凑SaCas9衍生物对DNA切割和碱基编辑的影响。

图7是dSaCas9变体的结构域组织的图(701)，其包括与FokI结构域融合的分割的dSaCas9和分割的mini-dSaCas9-4并且还包括融合至dSaCas9和mini-dSaCas9-4N末端的Fok1。

图8是证明DNA切割效率的EYFP重构测定的图(801)以及通过融合至分割的dSaCas9 或分割的minidSaCas9-4的二聚FokI核酸酶进行DNA切割的图示(803)。EYFP重构测定评估与FokI以及各自包含与靶互补的18-nt序列的两个截短gRNA融合的dSaCas9衍生物的 DNA切割效率。在一个说明性实施例中，当将FokI融合至mini-SaCas9-2的N-末端时，未检测到DNA切割活性。一个可能的原因是这种蛋白质结构中的FokI可能位于靶DNA的远端。但是，由于FokI核酸酶结构域的N-末端和C-末端之间的预测距离为这使得在dSaCas9的中间找到合适的插入位置具有挑战。在一个替代实施例中，将dSaCas9在残基733处分割，并在分割点之后利用一式三份的G-接头(“GGGGS”)融合FokI。在另一个说明性实施例中，将可能干扰两个分割片段重构的C-末端片段的前四个残基(△734-737)除去。没有HNH结构域的分裂的dSaCas9或分裂的mini-dSaCas9-4得到野生型SaCas9诱导的 EYFP表达水平的约8％至30％，其中在PAM外方向的间隔区在12bp至24bp的范围。

图9是间隔区长度为12-bp至24-bp的分割dSaCas9变体的DNA切割效率的图示(901)。间隔区长度为12-bp至24-bp的分割dSaCas9:FokI、分割mini-dSaCas9-4:FokI以及Fok1与 dSaCas9和mini-dSacas9-4的N末端融合的DNA切割效率。每个条形显示在HEK293细胞中转染后48小时通过流式细胞仪测量的EYFP荧光的平均倍数变化(平均值±SEM；n＝3)。

在接下来的说明性实施例中，进一步详细描述了用于转录激活的紧凑CRISPR/Cas系统的构建。

图10是VPR、VTR1、VTR2和VTR3转录激活结构域的示意图(1001)，以及通过使用EBFP2报告系统评估的相应基因激活效率的图示(1003)。在该说明性实施例中，设计了基于dCas9-VPR的紧凑转录激活因子。整个P65包含在N-末端的DNA结合结构域和在C- 末端的两个反式激活结构域(TA1和TA2)。但是，在由三部分组成的VPR结构域中仅包含 TA2和部分TA1。为了减小dCas9-VPR的尺寸，可以通过用TA1和TA2结构域替换VPR 中的P65结构域来构造mini-SaCas9-3:VTR1。在另一个说明性实施例中，用两个重复的TA1 结构域来构建VPR中P65结构域，并且称为VTR2。VTR1结构域和VTR2结构域分别保留了45％和21％的VPR结构域的反式激活效率。另外描述了VTR3，其比VTR2和VTR1短约200bp，从而使其成为最佳的说明性实施例。

图11是用于转录激活的SpyTag和MoonTag重复阵列的示意图(1101)，以及相应基因激活效率数据的图示(1103)。在说明性实施例中，通过使用利用优化的gRNA-2的EBFP2报告系统评估相应的基因激活效率。数据显示为在转染入HEK293细胞后72小时用流式细胞仪测量的EBFP2荧光的平均倍数变化(平均值±SEM；n＝3)。最近，已经显示来源于化脓性链球菌纤连蛋白结合蛋白(FbaB)的13个残基的肽标签(SpyTag)与被称为SpyCatcher 的116个残基的结合伴侣形成共价键。

在说明性实施例中，通过将SpyTag的四个串联重复序列融合至mini-SaCas9-3的C末端并将SpyCatcher与VPR结构域融合来构建尺寸比SunTag系统小的重复肽阵列，允许在细胞中自发组装VPR反式激活支架。与阴性对照相比，SpyTag系统将增强的蓝色荧光蛋白2(EBFP2)报告基因的表达诱导至100倍。在说明性实施例中，通过搜索SpyTag和SpyCatcher的同源物，从海豹链球菌(Streptococcus phocae)发现了与FbaB具有60％序列相似性的推定蛋白质(登录号WP_054278706)。在说明性实施例中，类似于SpyTag和SpyCatcher该蛋白质被分割。作为说明性实施例的直接和预期结果，本文公开了被称为MoonTag系统的类似支架系统，并且通过将13个残基的MoonTag的四个串联重复序列与mini-SaCas9-3融合并使MoonCatcher和VPR结构域形成复合体加以实现。尽管MoonTag系统与SpyTag系统不正交，但是MoonTag系统激活EBFP2表达的效率提高至5倍。

图12是通过使用装载有紧凑CRISPR/Cas9系统的单个AAV进行转录激活的示意图(1201)，以及EBFP2激活效率的图示(1203)。在说明性实例中，描绘了通过使用装载有紧凑CRISPR/Cas9系统的单个AAV的转录激活，所述紧凑CRISPR/Cas9系统包含 mini-SaCas9-3:VTR1和U6驱动的优化gRNA-2。通过瞬时转染然后AAV感染将编码EBFP2 报告基因和mKate2对照基因的质粒DNA引入至HEK293细胞。下方的图显示了在编码 mini-SaCas9-3:VTR1的AAV转染之后的EBFP2的激活效率和代表性显微图像(1205)。图像中的比例尺代表200μm。数据显示为转染和AAV感染之后72小时使用流式细胞仪测量的EBFP2荧光的平均倍数变化(平均值±SEM；n＝3)。在该说明性实施例中，产生了编码组成性表达的mini-SaCas9-3:VTR1和靶向TRE启动子的优化的gRNA的单个AAV病毒。通过瞬时转染，将TRE驱动的EBFP2报告基因引入至HEK293细胞。在说明性实施例中，与HEK293细胞中的阴性对照相比，AAV感染将EBFP2表达激活至高达～130倍，其也比装载有mini-SaCas9-3:VP64的AAV病毒更有效。

图13是将mini-Cas9、效应子结构域、gRNA表达盒和其它部分包装在单个AAV载体中以用于转录激活、DNA切割和碱基编辑的示意图(1301)。通过将说明性实施例与广泛的工作实例相结合，基于跨多种2类CRISPR效应子的结构信息，通过缺失保守的HNH和/ 或REC-C结构域工程化改造了一组紧凑Cas9衍生物。此外，还公开了通过分割mini-dSaCas9 并且在分割点正后方融合FokI结构域来工程化改造二聚gRNA指导的核酸酶的新策略，由于二聚FokI核酸酶与靶序列的接近度更近，该新策略可以提高目标DNA切割效率并潜在降低脱靶效率。通过将优化的紧凑gRNA表达盒与尺寸缩小的SaCas9衍生物组合，公开了将具有用于反式激活、DNA切割和碱基编辑的不同效应子结构域的整个CRISPR/Cas系统装载到单个AAV病毒中的实用方法。这样的仅用一个AAV-CRISPR/Cas9系统在需要安全有效的体内递送的生物医学应用中特别有吸引力。

图14是miniSaCas9-3和分割SaCas9的示意图(1401)以及相应miniSaCas9-3和分割 SaCas9激活效率的图示数据(1403)。

图15是示出了间隔区的CCR5基因的基因序列图示(1501)，以及示出了CCR5基因的相对切割百分比的凝胶电泳图示数据(1503)。

在描述中，阐述了许多具体细节以便提供对本发明实施例的透彻理解。然而，对于本领结构域的普通技术人员将显而易见的是，不需要采用具体细节来实践本实施例。在另一些情况下，没有详细描述公知的材料或方法，以避免使本发明的实施例不清楚。

在整个说明书中，对“一个实施例”、“实施例”，“一个实例”或“实例”的引用意味着结合该实施例或实例描述的特定特征、结构或特性包括在本发明的至少一个实施例中。因此，在整个说明书中各处出现的短语“在一个实施例中”、“在实施例中”、“一个实例”或“实例”不一定全部都指的是同一实施例或实例。此外，在一个或多个实施例或实例中，特定特征、结构或特性可以以任何合适的组合和/或子组合来组合。另外，应当理解，随此提供的附图是出于对本领结构域普通技术人员的解释目的，并且附图不一定按比例绘制。

如本文所使用的，术语“包括”、“包含”、“含有”、“含”、“具有”或其任何其它变体旨在覆盖非排他性的包含。例如，包括一系列元素的过程、制品或设备不一定仅限于那些元素，而是可以包括未明确列出或这类过程、制品或设备所固有的其它元素。

另外，本文给出的任何实例或示例均不应以任何方式视为对与它们一起使用的任何一个或多个术语的约束、限制或表达定义。相反，这些实例或示例应被视为是相对于一个特定实施例进行描述的，并且仅是示例性的。本领结构域普通技术人员将理解，与这些实例或示例一起使用的一个或多个术语将涵盖可能未与其一起或在说明书其它地方给出的其它实施例，并且所有这样的实施例旨在被包括在该一个或多个术语的范围内。指定这样的非限制性实例或示例的语言包括但不限于：“例如”、“诸如”、“如”和“在一个实施例中”。

工业适用性

要求保护的本发明在生物医学和工业生物技术应用中具有工业适用性。由于更高的特征性，改进的Cas9系统提供了比传统Cas9系统更强的基因编辑和调节控制能力。

序列表自由文本

本申请包含序列表，该序列表已经以ASCII格式提交，并且通过引用以其整体并入本文。所述ASCII副本创建于2017年4月28日，命名为ZX2seqlist_ST25.txt，大小为225KB。

本专利所涉及的序列如下。

序列表

Seq ID 1gRNAa TACGTTCTCTATCACTGATA

Seq ID 2gRNAb TACGTTCTCTATCACTGATA

Seq ID 3crRNAa CTCCCTATCAGTGATAGAGAACG

Seq ID 4gRNAc CGTTCTCTATCACTGATA

Seq ID 5gRNAd ACTAGAAATTCACCGAGC

Seq ID 6gRNA-15bp CTCACTCAACAGTGATAGAGA

Seq ID 7gRNA-12bp TTGCTCACTCAACAGTGATAG

Seq ID 8

dSpCas9:VPR

MPKKKRKVGGGSPGMDKKYSIGLAIGTNSVGWAVITDEYKVPS KKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYT RRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPI FGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMI KFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVD AKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNF KSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKN LSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDG TEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFY PFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITP WNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFT VYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVK QLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFL DNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQL KRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQT VKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMK RIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQ ELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDK AGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITL KSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKY PKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNF FKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMP QVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGF DSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPI DFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKG NELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYL DEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHL FTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYE TRIDLSQLGGDTSRADPKKKRKVEASGSGRADALDDFDLDMLG SDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRS QYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIA VPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASA LAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVA PPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDL ASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRP PDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSGSRDS REGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPL PASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDP DEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDE LTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLS IFDTSLF

Seq ID 9

mini-dSpCas9-1:VPR

MPKKKRKVGGGSPGMDKKYSIGLAIGTNSVGWAVITDEYKVPS KKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYT RRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPI FGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMI KFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVD AKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNF KSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKN LSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDG TEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFY PFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITP WNFEEVVDKGASAQSFIERMTNFDKAQVSGQGDSLHEHIANLA GSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQK GQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL QNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRS DKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTK AERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDEN DKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKAT AKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGR DFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIAR KKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELL GITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGR KRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNE QKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHR DKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGDTSRADPKKKRKVEASGSGRA DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDAL DDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPF SGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPT MVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAP VPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGAL LGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYP EAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDF SALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRP FHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAP AVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQM DLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDE CLLHAMHISTGLSIFDTSLF

Seq ID 10

mini-dSpCas9-2:VPR

MPKKKRKVGGGSPGMDKKYSIGLAIGTNSVGWAVITDEYKVPS KKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYT RRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPI FGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMI KFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVD AKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNF KSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKN LSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDG TEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFY PFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITP WNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFT VYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVK QLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFL DNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQL KRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQT VKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNITQRKFD NLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTK YDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEI GKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVW DKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDK LIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSV KELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFEL ENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSP EDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAY NKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTST KEVLDATLIHQSITGLYETRIDLSQLGGDTSRADPKKKRKVEASG SGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLG SDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIM KKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINY DEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALA QAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDE DLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPM LMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSI ADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQ PKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQP LDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAA ICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF

Seq ID 11

dSaCas9:VPR

MPKKKRKVGGGSPGKRNYILGLAIGITSVGYGIIDYETRDVIDAG VRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYN LLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVH NVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGE VRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLET RRTYYEGPGEGSPFGWKDIKEWYEMLMGHCTYFPEELRSVKYA YNADLYNALNDLNNLVITRDENEKLEYYEKFQIIENVFKQKKKP TLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARK EIIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLK GYTGTHNLSLKAINLILDELWHTNDNQIAIFNRLKLVPKKVDLS QQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELA REKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAKYLIEKI KLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSF NNKVLVKQEEASKKGNRTPFQYLSSSDSKISYETFKKHILNLAK GKGRISKTKKEYLLEERDINRFSVQKDFINRNLVDTRYATRGLM NLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKH HAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPE IETEQEYKEIFITPHQIKHIKDFKDYKYSHRVDKKPNRKLINDTLY STRKDDKGNTLIVNNLNGLYDKDNDKLKKLINKSPEKLLMYHH DPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGP VIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLD NGVYKFVTVKNLDVIKKENYYEVNSKCYEEAKKLKKISNQAEFI ASFYKNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYLENM NDKRPPHIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGT SRADPKKKRKVEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRI EEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKP APQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQA PAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEG TLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPG LPNGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAG SAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPV HEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALR EMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF

Seq ID 12

mini-SaCas9-1:VPR

MPKKKRKVGGGSPGKRNYILGLDIGITSVGYGIIDYETRDVIDAG VRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYN LLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVH NVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGE VRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLET RRTYYEGPGEGSPFGWKDIKEWYEMLGGGSGGGILSPVVKRSFI QSIKVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQ TNERIEEIIRTTGKENAKYLIEKIKLHDMQEGKCLYSLEAIPLEDL LNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQEENSKKGNRTPFQ YLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFS VQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGG FTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLD KAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIKDFK DYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYD KDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDITDD YPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENY YEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRVIGV NNDLLNRIEVNMIDITYREYLENMNDKRPPHIIKTIASKTQSIKKY STDILGNLYEVKSKKHPQIIKKGTSRADPKKKRKVEASGSGRAD ALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALD DFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFS GPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTM VFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPV PVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALL GNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPE AITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFS ALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPF HPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPA VTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMD LSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDEC LLHAMHISTGLSIFDTSLF

Seq ID 13

mini-SaCas9-2:VPR

MPKKKRKVGGGSPGKRNYILGLDIGITSVGYGIIDYETRDVIDAG VRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYN LLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVH NVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGE VRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLET RRTYYEGPGEGSPFGWKDIKEWYEMLKRRRRHRILSPVVKRSFI QSIKVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQ TNERIEEIIRTTGKENAKYLIEKIKLHDMQEGKCLYSLEAIPLEDL LNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQEENSKKGNRTPFQ YLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFS VQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGG FTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLD KAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIKDFK DYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYD KDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDITDD YPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENY YEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRVIGV NNDLLNRIEVNMIDITYREYLENMNDKRPPHIIKTIASKTQSIKKY STDILGNLYEVKSKKHPQIIKKGTSRADPKKKRKVEASGSGRAD ALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALD DFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFS GPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTM VFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPV PVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALL GNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPE AITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFS ALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPF HPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPA VTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMD LSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDEC LLHAMHISTGLSIFDTSLF

Seq ID 14

mini-SaCas9-3:VPR

MPKKKRKVGGGSPGKRNYILGLDIGITSVGYGIIDYETRDVIDAG VRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYN LLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVH NVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGE VRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLET RRTYYEGPGEGSPFGWKDIKEWYEMLKRRRRHRILSPVVKRSFI QSIKVINAIIKKYGLPNDIIIELGSKRYATRGLMNLLRSYFRVNNL DVKVKSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANAD FIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFIT PHQIKHIKDFKDYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTL IVNNLNGLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLI MEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGN KLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTV KNLDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYKNDLI KINGELYRVIGVNNDLLNRIEVNMIDITYREYLENMNDKRPPHII KTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGTSRADPKKK RKVEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDD FDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTY ETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTS SLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPA MVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQ LQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAP HTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSG DEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFE GREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLT PAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVI PQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTP ELNEILDTFLNDECLLHAMHISTGLSIFDTSLF

Seq ID 15

dAsCpf1:VPR

MPKKKRKVGGGSPGMTQFEGFTNLYQVSKTLRFELIPQGKTLK HIQEQGFIEEDKARNDHYKELKPIIDRIYKTYADQCLQLVQLDW ENLSAAIDSYRKEKTEETRNALIEEQATYRNAIHDYFIGRTDNLT DAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSF DKFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIF TRLITAVPSLREHFENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQI DLYNQLLGGISREAGTEKIKGLNEVLNLAIQKNDETAHIIASLPH RFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLRNENVLE TAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERR ISELTGKITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKT SEILSHAHAALDQPLPTTLKKQEEKEILKSQLDSLLGLYHLLDWF AVDESNEVDPEFSARLTGIKLEMEPSLSFYNKARNYATKKPYSV EKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQ KGRYKALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKA VTAHFQTHTTPILLSNNFIEPLEITKEIYDLNNPEKEPKKFQTAYA KKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDLSSLRPSSQY KDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNK DFAKGHHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPK SRMKRMAHRLGEKMLNKKLKDQKTPIPDTLYQELYDYVNHRL SHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFHVPITLNYQ AANSPSKFNQRVNAYLKEHPETPIIGIARGERNLIYITVIDSTGKIL EQRSLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQG YLSQVIHEIVDLMIHYQAVVVLENLNFGFKSKRTGIAEKAVYQQ FEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQFTSFAKMGT QSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFD FLHYDVKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNET QFDAKGTPFIAGKRIVPVIENHRFTGRYRDLYPANELIALLEEKGI VFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMRNSNAATGED YINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLL NHLKESKDLKLQNGISNQDWLAYIQELRNTSRADPKKKRKVEA SGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDM LGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSI MKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTIN YDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSAL AQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDD EDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEP MLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFS SIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVC QPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVP QPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEE AAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEI LDTFLNDECLLHAMHISTGLSIFDTSLF

Seq ID 16

mini-AsCpf1-1:VPR

MPKKKRKVGGGSPGMTQFEGFTNLYQVSKTLRFELIPQGKTLK HIQEQGFIEEDKARNDHYKELKPIIDRIYKTYADQCLQLVQLDW ENLSAAIDSYRKEKTEETRNALIEEQATYRNAIHDYFIGRTDNLT DAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSF DKFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIF TRLITAVPSLREHFENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQI DLYNQLLGGISREAGTEKIKGLNEVLNLAIQKNDETAHIIASLPH RFIPLFKQILSDRNTLSFILEEFGGGSYSVEKFKLNFQMPTLASGW DVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKALSFEPTEKTSE GFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNF IEPLEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKW IDFTRDFLSKYTKTTSIDLSSLRPSSQYKDLGEYYAELNPLLYHIS FQRIAEKEIMDAVETGKLYLFQIYNKDFAKGHHGKPNLHTLYW TGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLN KKLKDQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITK EVSHEIIKDRRFTSDKFFFHVPITLNYQAANSPSKFNQRVNAYLK EHPETPIIGIDRGERNLIYITVIDSTGKILEQRSLNTIQQFDYQKKL DNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQ AVVVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKD YPAEKVGGVLNPYQLTDQFTSFAKMGTQSGFLFYVPAPYTSKID PLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDVKTGDFILHFK MNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVP VIENHRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLEND DSHAIDTMVALIRSVLQMRNSNAATGEDYINSPVRDLNGVCFDS RFQNPEWPMDADANGAYHIALKGQLLLNHLKESKDLKLQNGIS NQDWLAYIQELRNTSRADPKKKRKVEASGSGRADALDDFDLD MLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLI NSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPP PRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQIS QASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAV FTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTG AQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSG SRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWA NRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRG HLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHI STGLSIFDTSLF

Seq ID 17

dSaCas9N:FokI

MPKKKRKVGGGSPGKRNYILGLAIGITSVGYGIIDYETRDVIDAG VRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYN LLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVH NVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGE VRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLET RRTYYEGPGEGSPFGWKDIKEWYEMLMGHCTYFPEELRSVKYA YNADLYNALNDLNNLVITRDENEKLEYYEKFQIIENVFKQKKKP TLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARK EIIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLK GYTGTHNLSLKAINLILDELWHTNDNQIAIFNRLKLVPKKVDLS QQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELA REKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAKYLIEKI KLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSF NNKVLVKQEEASKKGNRTPFQYLSSSDSKISYETFKKHILNLAK GKGRISKTKKEYLLEERDINRFSVQKDFINRNLVDTRYATRGLM NLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKH HAEDALIIANADFIFKEWKKLDKAKKVMENQMFGGGSGGGSG GGSLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEM KVMEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTK AYSGGYNLPIGQADEMQRYVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVLSVEELLIGG EMIKAGTLTLEEVRRKFNNGEINFTSGGGSKRPAATKKAGQAK KKKSR

Seq ID 18

dSaCas9C

MPKKKRKVGGGSPGAESMPEIETEQEYKEIFITPHQIKHIKDFKD YKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYDK DNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPL YKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDITDDY PNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRVIGVN NDLLNRIEVNMIDITYREYLENMNDKRPPHIIKTIASKTQSIKKYS TDILGNLYEVKSKKHPQIIKKGTSGGGSKRPAATKKAGQAKKK KSR

Seq ID 19

mini-dSaCas9-4N:FokI

MPKKKRKVGGGSPGKRNYILGLAIGITSVGYGIIDYETRDVIDAG VRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYN LLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVH NVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGE VRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLET RRTYYEGPGEGSPFGWKDIKEWYEMLMGHCTYFPEELRSVKYA YNADLYNALNDLNNLVITRDENEKLEYYEKFQIIENVFKQKKKP TLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARK EIIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLK GYTGTHNLSLKAINLILDELWHTNDNQIAIFNRLKLVPKKVDLS QQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELG SKRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKF KKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQM FGGGSGGGSGGGSLVKSELEEKKSELRHKLKYVPHEYIELIEIAR NSTQDRILEMKVMEFFMKVYGYRGKHLGGSRKPDGAIYTVGSP IDYGVIVDTKAYSGGYNLPIGQADEMQRYVEENQTRNKHINPNE WWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINFTSGGGSKRPAAT KKAGQAKKKKSR

Seq ID 20

AID:dSaCas9:UGI

MPKKKRKVGGGSPGMSSETGPVAVDPTLRRRIEPHEFEVFFDPR ELRKETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTER YFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLY HHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIA LQSCHYQRLPPHILWATGLKSGSETPGTSESATPEKRNYILGLAI GITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARR LKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQK LSEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSK ALEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKV QKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWY EMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENE KLEYYEKFQIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTST GKPEFTNLKVYHDIKDITARKEIIENAELLDQIAKILTIYQSSEDIQ EELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAINLILDELWHTN DNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSI KVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNE RIEEIIRTTGKENAKYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNN PFNYEVDHIIPRSVSFDNSFNNKVLVKQEEASKKGNRTPFQYLSS SDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFSVQK DFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSF LRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKK VMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIKDFKDYKY SHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYDKDND KLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKY YEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDITDDYPNSR NKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENYYEVNS KCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRVIGVNNDLL NRIEVNMIDITYREYLENMNDKRPPHIIKTIASKTQSIKKYSTDIL GNLYEVKSKKHPQIIKKGSGGSTNLSDIIEKETGKQLVIQESILML PEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSPPKKKRKV

Seq ID 21

AID:mini-dSaCas9-5:UGI

MPKKKRKVGGGSPGMSSETGPVAVDPTLRRRIEPHEFEVFFDPR ELRKETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTER YFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLY HHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIA LQSCHYQRLPPHILWATGLKSGSETPGTSESATPEKRNYILGLAI GITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARR LKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQK LSEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSK ALEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKV QKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWY EMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENE KLEYYEKFQIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTST GKPEFTNLKVYHDIKDITARKEIIENAELLDQIAKILTIYQSSEDIQ EELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAINLILDELWHTN DNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSI KVINAIIKKYGLPNDIIIELGSKRYATRGLMNLLRSYFRVNNLDV KVKSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIF KEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPH QIKHIKDFKDYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIV NNLNGLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIME QYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLN AHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNL DVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKING ELYRVIGVNNDLLNRIEVNMIDITYREYLENMNDKRPPHIIKTIAS KTQSIKKYSTDILGNLYEVKSKKHPQIIKKGSGGSTNLSDIIEKET GKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVML LTSDAPEYKPWALVIQDSNGENKIKMLSGGSPPKKKRKV

Seq ID 22

mini-dSaCas9-6N:AID

MPKKKRKVGGGSPGKRNYILGLAIGITSVGYGIIDYETRDVIDAG VRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYN LLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVH NVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGE VRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLET RRTYYEGPGEGSPFGWKDIKEWYEMLMGHCTYFPEELRSVKYA YNADLYNALNDLNNLVITRDENEKLEYYEKFQIIENVFKQKKKP TLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARK EIIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLK GYTGTHNLSLKAINLILDELWHTNDNQIAIFNRLKLVPKKVDLS QQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELA REKGGGSGGGSMSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRK ETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCP NTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHAD PRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWP RYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCH YQRLPPHILWATGLKSGSETPGTSESATPETSGGGSKRPAATKK AGQAKKKKSR

Seq ID 23

mini-dSaCas9-6C:UGI

MPKKKRKVGGGSPGTRYATRGLMNLLRSYFRVNNLDVKVKSI NGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWK KLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIK DFKDYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNG LYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDI TDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKK ENYYEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRV IGVNNDLLNRIEVNMIDITYREYLENMNDKRPPHIIKTIASKTQSI KKYSTDILGNLYEVKSKKHPQIIKKGSGGSTNLSDIIEKETGKQL VIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSD APEYKPWALVIQDSNGENKIKMLSGGSPPKKKRKV

Seq ID 24

VTR1

GSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDML GSDALDDFDLDMLINSALLQLQFDDEDLGALLGNSTDPAVFTDL ASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRP PDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISSGSGSG SRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWA NRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHL LEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRG HLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHI STGLSIFDTSLFTSGGGSKRPAATKKAGQAKKKKSR

Seq ID 25

VTR2

GSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDML GSDALDDFDLDMLINSRLVTGAQRPPDPAPAPLGAPGLPNGLLS GDEDFSSIADMDFSALLSQISSGGGSRLVTGAQRPPDPAPAPLGA PGLPNGLLSGDEDFSSIADMDFSALLSQISSGSGSGSRDSREGMF LPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLA PTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETS QAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTL ESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLFTSGGGSKRPAATKKAGQAKKKKSR

Seq ID 26

SpyCatcher:VPR

MPKKKRKVGGGSPGGAMVDTLSGLSSEQGQSGDMTIEEDSATH IKFSKRDEDGKELAGATMELRDSSGKTISTWISDGQVKDFYLYP GKYTFVETAAPDGYEVATAITFTVNEQGQVTVNGKATKGDAHI TGTSRADPKKKRKVEASGSGRADALDDFDLDMLGSDALDDFDL DMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASV PKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVL PQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQA GEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQ QLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKP EAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPT GPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVK ALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMT EDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF

Seq ID 27

4x SpyTag

GGGSGGAHIVMVDAYKPTKGGSGGGGSGGAHIVMVDAYKPTK GGSGGGGSGGAHIVMVDAYKPTKGGSGGGGSGGAHIVMVDAY KPTK

Seq ID 28

MoonCatcher:VPR

MPKKKRKVGGGSPGNHVIETEQNLPNEDGQSGNIIEQEDSKTLV KFSKRDIKGNELAGATIELRDLSGKSIQSWVSDGKAKDFYLLPG SYEFVETAAPEGYQIATKIMFTISTDGRITVDGQLVTGTGSGRAD ALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALD DFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFS GPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTM VFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPV PVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALL GNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPE AITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFS ALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPF HPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPA VTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMD LSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDEC LLHAMHISTGLSIFDTSLF

Seq ID 29

4x MoonTag

GGGSGGAHIVMVDNYKPIVGGSGGAHIVMVDNYKPIVGGSGGA HIVMVDNYKPIVGGSGGGGSGGAHIVMVDNYKPIV

Seq ID 30

VTR3序列

GSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINS RLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISSGGGSRLVTGAQR PPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISSGSGSQPLDPAPAVTPEASHLL EDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNL DSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLFTS

Seq ID 31

gRNA-ccr5-1

GCTGTGTTTGCGTCTCTCCC

Seq ID 32

gRNA-ccr5-2

GGGGTGGTGACAAGTGTGAT

Seq ID 33

Mini-saCas9-4:VPR

MPKKKRKVGGGSPGGGGSKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEAN VENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKL SEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKK DGEVRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPF GWKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEK FQIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEIIEN AELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAINLILDELW HTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPN DIIIELGSKRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKH HAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIK HIKDFKDYKYSHRVDKKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYDKDNDKLKKLIN KSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKI KYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKEN YYEVNSKCYEEAKKLKKISNQAEFIASFYKNDLIKINGELYRVIGVNNDLLNRIEVNMIDIT YREYLENMNDKRPPHIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGTSRADPKKK RKVEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDL DMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASV PKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALA QAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTD LASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNG LLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPF HPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQ AVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF

Seq ID 34

Mini-saCas9-5:VPR

MPKKKRKVGGGSPGGGGSKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEAN VENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKL SEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKK DGEVRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPF GWKDIKEWYEMLMGHCTKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGL PNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAKYLIEKIKLHDMQEG KCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQEENSKKGNRTPFQYLSS SDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFSVQKDFINRNLVDTRYATRGL MNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFK EWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIKDFKDYKYSHRVD KKPNRKLINDTLYSTRKDDKGNTLIVNNLNGLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLNAHLDITD DYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENYYEVNSKCYEEAKKL KKISNQAEFIASFYKNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYLENMNDKRPPH IIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGTSRADPKKKRKVEASGSGRADALD DFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLINSRSQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTI NYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQ AVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQ GIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDF SALLGSGSGSRDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASL APTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQ KEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHA MHISTGLSIFDTSLF

Seq ID 35

GSK接头

GSK。

序列表

<110> 清华大学

<120> 工程化改造用于由增强的指导RNA优化的基因编辑和转录调节的最小化SaCas9CRISPR/Cas系统

<130> PIDC3171800PCN

<160> 34

<170> PatentIn version 3.5

<210> 1

<211> 20

<212> DNA

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 1

tacgttctct atcactgata 20

<210> 2

<211> 20

<212> DNA

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 2

tacgttctct atcactgata 20

<210> 3

<211> 23

<212> DNA

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 3

ctccctatca gtgatagaga acg 23

<210> 4

<211> 18

<212> DNA

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 4

cgttctctat cactgata 18

<210> 5

<211> 18

<212> DNA

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 5

actagaaatt caccgagc 18

<210> 6

<211> 21

<212> DNA

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 6

ctcactcaac agtgatagag a 21

<210> 7

<211> 21

<212> DNA

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 7

ttgctcactc aacagtgata g 21

<210> 8

<211> 1913

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 8

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Met Asp

1 5 10 15

Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp

20 25 30

Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val

35 40 45

Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala

50 55 60

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

65 70 75 80

Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu

85 90 95

Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe

100 105 110

His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu

115 120 125

Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu

130 135 140

Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr

145 150 155 160

Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile

165 170 175

Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn

180 185 190

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

195 200 205

Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala

210 215 220

Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile

225 230 235 240

Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile

245 250 255

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

260 265 270

Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp

275 280 285

Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe

290 295 300

Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu

305 310 315 320

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

325 330 335

Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu

340 345 350

Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln

355 360 365

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

370 375 380

Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr

385 390 395 400

Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln

405 410 415

Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu

420 425 430

Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys

435 440 445

Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr

450 455 460

Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr

465 470 475 480

Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val

485 490 495

Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe

500 505 510

Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu Leu

515 520 525

Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val

530 535 540

Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys

545 550 555 560

Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys

565 570 575

Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val

580 585 590

Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr

595 600 605

His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu

610 615 620

Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe

625 630 635 640

Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu

645 650 655

Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr Gly

660 665 670

Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys Gln

675 680 685

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

690 695 700

Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu

705 710 715 720

Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu

725 730 735

His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu

740 745 750

Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg His

755 760 765

Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr

770 775 780

Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu

785 790 795 800

Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu

805 810 815

Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn

820 825 830

Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser

835 840 845

Asp Tyr Asp Val Asp Ala Ile Val Pro Gln Ser Phe Leu Lys Asp Asp

850 855 860

Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys

865 870 875 880

Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr

885 890 895

Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp

900 905 910

Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala

915 920 925

Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His

930 935 940

Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn

945 950 955 960

Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu

965 970 975

Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu Ile

980 985 990

Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val Val Gly

995 1000 1005

Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe Val

1010 1015 1020

Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys

1025 1030 1035

Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr

1040 1045 1050

Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn

1055 1060 1065

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

1070 1075 1080

Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val Arg

1085 1090 1095

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

1100 1105 1110

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

1115 1120 1125

Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro Lys

1130 1135 1140

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

1145 1150 1155

Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys Ser

1160 1165 1170

Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser Phe

1175 1180 1185

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

1190 1195 1200

Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe

1205 1210 1215

Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly Glu

1220 1225 1230

Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val Asn

1235 1240 1245

Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser Pro

1250 1255 1260

Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys His

1265 1270 1275

Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg

1280 1285 1290

Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala Tyr

1295 1300 1305

Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile

1310 1315 1320

Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe

1325 1330 1335

Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr

1340 1345 1350

Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly

1355 1360 1365

Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp Thr

1370 1375 1380

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

1385 1390 1395

Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu

1400 1405 1410

Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser

1415 1420 1425

Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala

1430 1435 1440

Leu Asp Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg Ser Gln

1445 1450 1455

Tyr Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg

1460 1465 1470

Lys Arg Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro

1475 1480 1485

Phe Ser Gly Pro Thr Asp Pro Arg Pro Pro Pro Arg Arg Ile Ala

1490 1495 1500

Val Pro Ser Arg Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln

1505 1510 1515

Pro Tyr Pro Phe Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu

1520 1525 1530

Phe Pro Thr Met Val Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser

1535 1540 1545

Ala Leu Ala Pro Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala

1550 1555 1560

Pro Ala Pro Ala Pro Ala Met Val Ser Ala Leu Ala Gln Ala Pro

1565 1570 1575

Ala Pro Val Pro Val Leu Ala Pro Gly Pro Pro Gln Ala Val Ala

1580 1585 1590

Pro Pro Ala Pro Lys Pro Thr Gln Ala Gly Glu Gly Thr Leu Ser

1595 1600 1605

Glu Ala Leu Leu Gln Leu Gln Phe Asp Asp Glu Asp Leu Gly Ala

1610 1615 1620

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

1625 1630 1635

Ser Val Asp Asn Ser Glu Phe Gln Gln Leu Leu Asn Gln Gly Ile

1640 1645 1650

Pro Val Ala Pro His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro

1655 1660 1665

Glu Ala Ile Thr Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp

1670 1675 1680

Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu

1685 1690 1695

Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe

1700 1705 1710

Ser Ala Leu Leu Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu

1715 1720 1725

Gly Met Phe Leu Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp

1730 1735 1740

Val Phe Glu Gly Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro

1745 1750 1755

Phe His Pro Pro Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala

1760 1765 1770

Ser Leu Ala Pro Thr Pro Thr Gly Pro Val His Glu Pro Val Gly

1775 1780 1785

Ser Leu Thr Pro Ala Pro Val Pro Gln Pro Leu Asp Pro Ala Pro

1790 1795 1800

Ala Val Thr Pro Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu

1805 1810 1815

Glu Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr

1820 1825 1830

Val Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp

1835 1840 1845

Leu Ser His Pro Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr

1850 1855 1860

Thr Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu

1865 1870 1875

Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu

1880 1885 1890

Cys Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe

1895 1900 1905

Asp Thr Ser Leu Phe

1910

<210> 9

<211> 1703

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 9

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Met Asp

1 5 10 15

Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp

20 25 30

Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val

35 40 45

Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala

50 55 60

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

65 70 75 80

Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu

85 90 95

Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe

100 105 110

His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu

115 120 125

Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu

130 135 140

Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr

145 150 155 160

Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile

165 170 175

Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn

180 185 190

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

195 200 205

Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala

210 215 220

Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile

225 230 235 240

Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile

245 250 255

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

260 265 270

Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp

275 280 285

Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe

290 295 300

Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu

305 310 315 320

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

325 330 335

Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu

340 345 350

Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln

355 360 365

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

370 375 380

Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr

385 390 395 400

Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln

405 410 415

Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu

420 425 430

Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys

435 440 445

Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr

450 455 460

Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr

465 470 475 480

Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val

485 490 495

Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe

500 505 510

Asp Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu His Ile

515 520 525

Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr

530 535 540

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

545 550 555 560

Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys

565 570 575

Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile

580 585 590

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

595 600 605

Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg

610 615 620

Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr

625 630 635 640

Asp Val Asp Ala Ile Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile

645 650 655

Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp

660 665 670

Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg

675 680 685

Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu

690 695 700

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

705 710 715 720

Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala

725 730 735

Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys

740 745 750

Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser

755 760 765

Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu Ile Asn Asn

770 775 780

Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val Val Gly Thr Ala

785 790 795 800

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

805 810 815

Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys Ser Glu Gln Glu

820 825 830

Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr Ser Asn Ile Met Asn

835 840 845

Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg

850 855 860

Pro Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu Ile Val Trp Asp Lys

865 870 875 880

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

885 890 895

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

900 905 910

Ser Ile Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys

915 920 925

Asp Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala

930 935 940

Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys

945 950 955 960

Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser

965 970 975

Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys

980 985 990

Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe

995 1000 1005

Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly Glu

1010 1015 1020

Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val Asn

1025 1030 1035

Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser Pro

1040 1045 1050

Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys His

1055 1060 1065

Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg

1070 1075 1080

Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala Tyr

1085 1090 1095

Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile

1100 1105 1110

Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe

1115 1120 1125

Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr

1130 1135 1140

Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly

1145 1150 1155

Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp Thr

1160 1165 1170

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

1175 1180 1185

Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu

1190 1195 1200

Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser

1205 1210 1215

Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala

1220 1225 1230

Leu Asp Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg Ser Gln

1235 1240 1245

Tyr Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg

1250 1255 1260

Lys Arg Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro

1265 1270 1275

Phe Ser Gly Pro Thr Asp Pro Arg Pro Pro Pro Arg Arg Ile Ala

1280 1285 1290

Val Pro Ser Arg Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln

1295 1300 1305

Pro Tyr Pro Phe Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu

1310 1315 1320

Phe Pro Thr Met Val Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser

1325 1330 1335

Ala Leu Ala Pro Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala

1340 1345 1350

Pro Ala Pro Ala Pro Ala Met Val Ser Ala Leu Ala Gln Ala Pro

1355 1360 1365

Ala Pro Val Pro Val Leu Ala Pro Gly Pro Pro Gln Ala Val Ala

1370 1375 1380

Pro Pro Ala Pro Lys Pro Thr Gln Ala Gly Glu Gly Thr Leu Ser

1385 1390 1395

Glu Ala Leu Leu Gln Leu Gln Phe Asp Asp Glu Asp Leu Gly Ala

1400 1405 1410

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

1415 1420 1425

Ser Val Asp Asn Ser Glu Phe Gln Gln Leu Leu Asn Gln Gly Ile

1430 1435 1440

Pro Val Ala Pro His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro

1445 1450 1455

Glu Ala Ile Thr Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp

1460 1465 1470

Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu

1475 1480 1485

Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe

1490 1495 1500

Ser Ala Leu Leu Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu

1505 1510 1515

Gly Met Phe Leu Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp

1520 1525 1530

Val Phe Glu Gly Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro

1535 1540 1545

Phe His Pro Pro Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala

1550 1555 1560

Ser Leu Ala Pro Thr Pro Thr Gly Pro Val His Glu Pro Val Gly

1565 1570 1575

Ser Leu Thr Pro Ala Pro Val Pro Gln Pro Leu Asp Pro Ala Pro

1580 1585 1590

Ala Val Thr Pro Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu

1595 1600 1605

Glu Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr

1610 1615 1620

Val Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp

1625 1630 1635

Leu Ser His Pro Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr

1640 1645 1650

Thr Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu

1655 1660 1665

Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu

1670 1675 1680

Cys Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe

1685 1690 1695

Asp Thr Ser Leu Phe

1700

<210> 10

<211> 1798

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 10

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Met Asp

1 5 10 15

Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp

20 25 30

Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val

35 40 45

Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala

50 55 60

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

65 70 75 80

Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu

85 90 95

Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe

100 105 110

His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu

115 120 125

Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu

130 135 140

Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr

145 150 155 160

Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile

165 170 175

Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn

180 185 190

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

195 200 205

Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala

210 215 220

Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile

225 230 235 240

Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile

245 250 255

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

260 265 270

Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp

275 280 285

Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe

290 295 300

Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu

305 310 315 320

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

325 330 335

Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu

340 345 350

Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln

355 360 365

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

370 375 380

Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr

385 390 395 400

Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln

405 410 415

Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu

420 425 430

Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys

435 440 445

Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr

450 455 460

Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr

465 470 475 480

Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val

485 490 495

Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe

500 505 510

Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu Leu

515 520 525

Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val

530 535 540

Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys

545 550 555 560

Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys

565 570 575

Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val

580 585 590

Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr

595 600 605

His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu

610 615 620

Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe

625 630 635 640

Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu

645 650 655

Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr Gly

660 665 670

Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys Gln

675 680 685

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

690 695 700

Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu

705 710 715 720

Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu

725 730 735

His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu

740 745 750

Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg His

755 760 765

Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr

770 775 780

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

785 790 795 800

Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile

805 810 815

Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln

820 825 830

Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu

835 840 845

Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp

850 855 860

Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu Ile Asn Asn Tyr

865 870 875 880

His His Ala His Asp Ala Tyr Leu Asn Ala Val Val Gly Thr Ala Leu

885 890 895

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

900 905 910

Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys Ser Glu Gln Glu Ile

915 920 925

Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe

930 935 940

Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro

945 950 955 960

Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly

965 970 975

Arg Asp Phe Ala Thr Val Arg Lys Val Leu Ser Met Pro Gln Val Asn

980 985 990

Ile Val Lys Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser

995 1000 1005

Ile Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys

1010 1015 1020

Asp Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val

1025 1030 1035

Ala Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser

1040 1045 1050

Lys Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met

1055 1060 1065

Glu Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala

1070 1075 1080

Lys Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro

1085 1090 1095

Lys Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu

1100 1105 1110

Ala Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro

1115 1120 1125

Ser Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys

1130 1135 1140

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

1145 1150 1155

Glu Gln His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser

1160 1165 1170

Glu Phe Ser Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys

1175 1180 1185

Val Leu Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu

1190 1195 1200

Gln Ala Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly

1205 1210 1215

Ala Pro Ala Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys

1220 1225 1230

Arg Tyr Thr Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His

1235 1240 1245

Gln Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln

1250 1255 1260

Leu Gly Gly Asp Thr Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys

1265 1270 1275

Val Glu Ala Ser Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe

1280 1285 1290

Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu

1295 1300 1305

Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met

1310 1315 1320

Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Ile

1325 1330 1335

Asn Ser Arg Ser Gln Tyr Leu Pro Asp Thr Asp Asp Arg His Arg

1340 1345 1350

Ile Glu Glu Lys Arg Lys Arg Thr Tyr Glu Thr Phe Lys Ser Ile

1355 1360 1365

Met Lys Lys Ser Pro Phe Ser Gly Pro Thr Asp Pro Arg Pro Pro

1370 1375 1380

Pro Arg Arg Ile Ala Val Pro Ser Arg Ser Ser Ala Ser Val Pro

1385 1390 1395

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

1400 1405 1410

Ile Asn Tyr Asp Glu Phe Pro Thr Met Val Phe Pro Ser Gly Gln

1415 1420 1425

Ile Ser Gln Ala Ser Ala Leu Ala Pro Ala Pro Pro Gln Val Leu

1430 1435 1440

Pro Gln Ala Pro Ala Pro Ala Pro Ala Pro Ala Met Val Ser Ala

1445 1450 1455

Leu Ala Gln Ala Pro Ala Pro Val Pro Val Leu Ala Pro Gly Pro

1460 1465 1470

Pro Gln Ala Val Ala Pro Pro Ala Pro Lys Pro Thr Gln Ala Gly

1475 1480 1485

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

1490 1495 1500

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

1505 1510 1515

Phe Thr Asp Leu Ala Ser Val Asp Asn Ser Glu Phe Gln Gln Leu

1520 1525 1530

Leu Asn Gln Gly Ile Pro Val Ala Pro His Thr Thr Glu Pro Met

1535 1540 1545

Leu Met Glu Tyr Pro Glu Ala Ile Thr Arg Leu Val Thr Gly Ala

1550 1555 1560

Gln Arg Pro Pro Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly

1565 1570 1575

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

1580 1585 1590

Ala Asp Met Asp Phe Ser Ala Leu Leu Gly Ser Gly Ser Gly Ser

1595 1600 1605

Arg Asp Ser Arg Glu Gly Met Phe Leu Pro Lys Pro Glu Ala Gly

1610 1615 1620

Ser Ala Ile Ser Asp Val Phe Glu Gly Arg Glu Val Cys Gln Pro

1625 1630 1635

Lys Arg Ile Arg Pro Phe His Pro Pro Gly Ser Pro Trp Ala Asn

1640 1645 1650

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

1655 1660 1665

His Glu Pro Val Gly Ser Leu Thr Pro Ala Pro Val Pro Gln Pro

1670 1675 1680

Leu Asp Pro Ala Pro Ala Val Thr Pro Glu Ala Ser His Leu Leu

1685 1690 1695

Glu Asp Pro Asp Glu Glu Thr Ser Gln Ala Val Lys Ala Leu Arg

1700 1705 1710

Glu Met Ala Asp Thr Val Ile Pro Gln Lys Glu Glu Ala Ala Ile

1715 1720 1725

Cys Gly Gln Met Asp Leu Ser His Pro Pro Pro Arg Gly His Leu

1730 1735 1740

Asp Glu Leu Thr Thr Thr Leu Glu Ser Met Thr Glu Asp Leu Asn

1745 1750 1755

Leu Asp Ser Pro Leu Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr

1760 1765 1770

Phe Leu Asn Asp Glu Cys Leu Leu His Ala Met His Ile Ser Thr

1775 1780 1785

Gly Leu Ser Ile Phe Asp Thr Ser Leu Phe

1790 1795

<210> 11

<211> 1597

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 11

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Lys Arg

1 5 10 15

Asn Tyr Ile Leu Gly Leu Ala Ile Gly Ile Thr Ser Val Gly Tyr Gly

20 25 30

Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly Val Arg Leu

35 40 45

Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg Ser Lys Arg

50 55 60

Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile Gln Arg Val

65 70 75 80

Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His Ser Glu Leu

85 90 95

Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu Ser Gln Lys

100 105 110

Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu Ala Lys Arg

115 120 125

Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr Gly Asn Glu

130 135 140

Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala Leu Glu Glu

145 150 155 160

Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys Asp Gly Glu

165 170 175

Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr Val Lys Glu

180 185 190

Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln Leu Asp Gln

195 200 205

Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg Arg Thr Tyr

210 215 220

Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys Asp Ile Lys

225 230 235 240

Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe Pro Glu Glu

245 250 255

Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr Asn Ala Leu

260 265 270

Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn Glu Lys Leu

275 280 285

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

290 295 300

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

305 310 315 320

Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys Pro Glu Phe

325 330 335

Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr Ala Arg Lys

340 345 350

Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala Lys Ile Leu

355 360 365

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

370 375 380

Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser Asn Leu Lys

385 390 395 400

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

405 410 415

Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala Ile Phe Asn

420 425 430

Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln Gln Lys Glu

435 440 445

Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro Val Val Lys

450 455 460

Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile Ile Lys Lys

465 470 475 480

Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg Glu Lys Asn

485 490 495

Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys Arg Asn Arg

500 505 510

Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr Gly Lys Glu

515 520 525

Asn Ala Lys Tyr Leu Ile Glu Lys Ile Lys Leu His Asp Met Gln Glu

530 535 540

Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu Asp Leu Leu

545 550 555 560

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

565 570 575

Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys Gln Glu Glu

580 585 590

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

595 600 605

Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile Leu Asn Leu

610 615 620

Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu Tyr Leu Leu

625 630 635 640

Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp Phe Ile Asn

645 650 655

Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu Met Asn Leu

660 665 670

Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys Val Lys Ser

675 680 685

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

690 695 700

Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp Ala Leu Ile

705 710 715 720

Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys Leu Asp Lys

725 730 735

Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys Gln Ala Glu

740 745 750

Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu Ile Phe Ile

755 760 765

Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp Tyr Lys Tyr

770 775 780

Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile Asn Asp Thr

785 790 795 800

Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu Ile Val Asn

805 810 815

Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu Lys Lys Leu

820 825 830

Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His Asp Pro Gln

835 840 845

Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly Asp Glu Lys

850 855 860

Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr Leu Thr Lys

865 870 875 880

Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile Lys Tyr Tyr

885 890 895

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

900 905 910

Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr Arg Phe Asp

915 920 925

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

930 935 940

Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser Lys Cys Tyr

945 950 955 960

Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala Glu Phe Ile

965 970 975

Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly Glu Leu Tyr

980 985 990

Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg Ile Glu Val Asn

995 1000 1005

Met Ile Asp Ile Thr Tyr Arg Glu Tyr Leu Glu Asn Met Asn Asp

1010 1015 1020

Lys Arg Pro Pro His Ile Ile Lys Thr Ile Ala Ser Lys Thr Gln

1025 1030 1035

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

1040 1045 1050

Val Lys Ser Lys Lys His Pro Gln Ile Ile Lys Lys Gly Thr Ser

1055 1060 1065

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

1070 1075 1080

Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly

1085 1090 1095

Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp

1100 1105 1110

Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu

1115 1120 1125

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

1130 1135 1140

Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg Lys

1145 1150 1155

Arg Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro Phe

1160 1165 1170

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

1175 1180 1185

Pro Ser Arg Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln Pro

1190 1195 1200

Tyr Pro Phe Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu Phe

1205 1210 1215

Pro Thr Met Val Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser Ala

1220 1225 1230

Leu Ala Pro Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala Pro

1235 1240 1245

Ala Pro Ala Pro Ala Met Val Ser Ala Leu Ala Gln Ala Pro Ala

1250 1255 1260

Pro Val Pro Val Leu Ala Pro Gly Pro Pro Gln Ala Val Ala Pro

1265 1270 1275

Pro Ala Pro Lys Pro Thr Gln Ala Gly Glu Gly Thr Leu Ser Glu

1280 1285 1290

Ala Leu Leu Gln Leu Gln Phe Asp Asp Glu Asp Leu Gly Ala Leu

1295 1300 1305

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

1310 1315 1320

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

1325 1330 1335

Val Ala Pro His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro Glu

1340 1345 1350

Ala Ile Thr Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp Pro

1355 1360 1365

Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu Leu

1370 1375 1380

Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe Ser

1385 1390 1395

Ala Leu Leu Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu Gly

1400 1405 1410

Met Phe Leu Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp Val

1415 1420 1425

Phe Glu Gly Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro Phe

1430 1435 1440

His Pro Pro Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala Ser

1445 1450 1455

Leu Ala Pro Thr Pro Thr Gly Pro Val His Glu Pro Val Gly Ser

1460 1465 1470

Leu Thr Pro Ala Pro Val Pro Gln Pro Leu Asp Pro Ala Pro Ala

1475 1480 1485

Val Thr Pro Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu Glu

1490 1495 1500

Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr Val

1505 1510 1515

Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp Leu

1520 1525 1530

Ser His Pro Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr Thr

1535 1540 1545

Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu Thr

1550 1555 1560

Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu Cys

1565 1570 1575

Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe Asp

1580 1585 1590

Thr Ser Leu Phe

1595

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<220>

<223> 通过合理设计产生的合成构建体

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Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Lys Arg

1 5 10 15

Asn Tyr Ile Leu Gly Leu Asp Ile Gly Ile Thr Ser Val Gly Tyr Gly

20 25 30

Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly Val Arg Leu

35 40 45

Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg Ser Lys Arg

50 55 60

Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile Gln Arg Val

65 70 75 80

Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His Ser Glu Leu

85 90 95

Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu Ser Gln Lys

100 105 110

Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu Ala Lys Arg

115 120 125

Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr Gly Asn Glu

130 135 140

Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala Leu Glu Glu

145 150 155 160

Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys Asp Gly Glu

165 170 175

Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr Val Lys Glu

180 185 190

Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln Leu Asp Gln

195 200 205

Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg Arg Thr Tyr

210 215 220

Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys Asp Ile Lys

225 230 235 240

Glu Trp Tyr Glu Met Leu Gly Gly Gly Ser Gly Gly Gly Ile Leu Ser

245 250 255

Pro Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala

260 265 270

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

275 280 285

Arg Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln

290 295 300

Lys Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr

305 310 315 320

Thr Gly Lys Glu Asn Ala Lys Tyr Leu Ile Glu Lys Ile Lys Leu His

325 330 335

Asp Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu

340 345 350

Glu Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile

355 360 365

Pro Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val

370 375 380

Lys Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr

385 390 395 400

Leu Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His

405 410 415

Ile Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys

420 425 430

Glu Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys

435 440 445

Asp Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly

450 455 460

Leu Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val

465 470 475 480

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

485 490 495

Trp Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu

500 505 510

Asp Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys

515 520 525

Lys Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu

530 535 540

Lys Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys

545 550 555 560

Glu Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys

565 570 575

Asp Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu

580 585 590

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

595 600 605

Leu Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys

610 615 620

Leu Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His

625 630 635 640

His Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr

645 650 655

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

660 665 670

Tyr Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys

675 680 685

Ile Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp

690 695 700

Asp Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro

705 710 715 720

Tyr Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr

725 730 735

Val Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn

740 745 750

Ser Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln

755 760 765

Ala Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn

770 775 780

Gly Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg

785 790 795 800

Ile Glu Val Asn Met Ile Asp Ile Thr Tyr Arg Glu Tyr Leu Glu Asn

805 810 815

Met Asn Asp Lys Arg Pro Pro His Ile Ile Lys Thr Ile Ala Ser Lys

820 825 830

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

835 840 845

Glu Val Lys Ser Lys Lys His Pro Gln Ile Ile Lys Lys Gly Thr Ser

850 855 860

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

865 870 875 880

Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp

885 890 895

Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp

900 905 910

Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp

915 920 925

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

930 935 940

Asp Arg His Arg Ile Glu Glu Lys Arg Lys Arg Thr Tyr Glu Thr Phe

945 950 955 960

Lys Ser Ile Met Lys Lys Ser Pro Phe Ser Gly Pro Thr Asp Pro Arg

965 970 975

Pro Pro Pro Arg Arg Ile Ala Val Pro Ser Arg Ser Ser Ala Ser Val

980 985 990

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

995 1000 1005

Ile Asn Tyr Asp Glu Phe Pro Thr Met Val Phe Pro Ser Gly Gln

1010 1015 1020

Ile Ser Gln Ala Ser Ala Leu Ala Pro Ala Pro Pro Gln Val Leu

1025 1030 1035

Pro Gln Ala Pro Ala Pro Ala Pro Ala Pro Ala Met Val Ser Ala

1040 1045 1050

Leu Ala Gln Ala Pro Ala Pro Val Pro Val Leu Ala Pro Gly Pro

1055 1060 1065

Pro Gln Ala Val Ala Pro Pro Ala Pro Lys Pro Thr Gln Ala Gly

1070 1075 1080

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

1085 1090 1095

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

1100 1105 1110

Phe Thr Asp Leu Ala Ser Val Asp Asn Ser Glu Phe Gln Gln Leu

1115 1120 1125

Leu Asn Gln Gly Ile Pro Val Ala Pro His Thr Thr Glu Pro Met

1130 1135 1140

Leu Met Glu Tyr Pro Glu Ala Ile Thr Arg Leu Val Thr Gly Ala

1145 1150 1155

Gln Arg Pro Pro Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly

1160 1165 1170

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

1175 1180 1185

Ala Asp Met Asp Phe Ser Ala Leu Leu Gly Ser Gly Ser Gly Ser

1190 1195 1200

Arg Asp Ser Arg Glu Gly Met Phe Leu Pro Lys Pro Glu Ala Gly

1205 1210 1215

Ser Ala Ile Ser Asp Val Phe Glu Gly Arg Glu Val Cys Gln Pro

1220 1225 1230

Lys Arg Ile Arg Pro Phe His Pro Pro Gly Ser Pro Trp Ala Asn

1235 1240 1245

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

1250 1255 1260

His Glu Pro Val Gly Ser Leu Thr Pro Ala Pro Val Pro Gln Pro

1265 1270 1275

Leu Asp Pro Ala Pro Ala Val Thr Pro Glu Ala Ser His Leu Leu

1280 1285 1290

Glu Asp Pro Asp Glu Glu Thr Ser Gln Ala Val Lys Ala Leu Arg

1295 1300 1305

Glu Met Ala Asp Thr Val Ile Pro Gln Lys Glu Glu Ala Ala Ile

1310 1315 1320

Cys Gly Gln Met Asp Leu Ser His Pro Pro Pro Arg Gly His Leu

1325 1330 1335

Asp Glu Leu Thr Thr Thr Leu Glu Ser Met Thr Glu Asp Leu Asn

1340 1345 1350

Leu Asp Ser Pro Leu Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr

1355 1360 1365

Phe Leu Asn Asp Glu Cys Leu Leu His Ala Met His Ile Ser Thr

1370 1375 1380

Gly Leu Ser Ile Phe Asp Thr Ser Leu Phe

1385 1390

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<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 13

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Lys Arg

1 5 10 15

Asn Tyr Ile Leu Gly Leu Asp Ile Gly Ile Thr Ser Val Gly Tyr Gly

20 25 30

Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly Val Arg Leu

35 40 45

Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg Ser Lys Arg

50 55 60

Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile Gln Arg Val

65 70 75 80

Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His Ser Glu Leu

85 90 95

Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu Ser Gln Lys

100 105 110

Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu Ala Lys Arg

115 120 125

Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr Gly Asn Glu

130 135 140

Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala Leu Glu Glu

145 150 155 160

Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys Asp Gly Glu

165 170 175

Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr Val Lys Glu

180 185 190

Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln Leu Asp Gln

195 200 205

Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg Arg Thr Tyr

210 215 220

Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys Asp Ile Lys

225 230 235 240

Glu Trp Tyr Glu Met Leu Lys Arg Arg Arg Arg His Arg Ile Leu Ser

245 250 255

Pro Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala

260 265 270

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

275 280 285

Arg Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln

290 295 300

Lys Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr

305 310 315 320

Thr Gly Lys Glu Asn Ala Lys Tyr Leu Ile Glu Lys Ile Lys Leu His

325 330 335

Asp Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu

340 345 350

Glu Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile

355 360 365

Pro Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val

370 375 380

Lys Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr

385 390 395 400

Leu Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His

405 410 415

Ile Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys

420 425 430

Glu Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys

435 440 445

Asp Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly

450 455 460

Leu Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val

465 470 475 480

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

485 490 495

Trp Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu

500 505 510

Asp Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys

515 520 525

Lys Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu

530 535 540

Lys Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys

545 550 555 560

Glu Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys

565 570 575

Asp Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu

580 585 590

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

595 600 605

Leu Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys

610 615 620

Leu Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His

625 630 635 640

His Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr

645 650 655

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

660 665 670

Tyr Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys

675 680 685

Ile Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp

690 695 700

Asp Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro

705 710 715 720

Tyr Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr

725 730 735

Val Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn

740 745 750

Ser Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln

755 760 765

Ala Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn

770 775 780

Gly Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg

785 790 795 800

Ile Glu Val Asn Met Ile Asp Ile Thr Tyr Arg Glu Tyr Leu Glu Asn

805 810 815

Met Asn Asp Lys Arg Pro Pro His Ile Ile Lys Thr Ile Ala Ser Lys

820 825 830

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

835 840 845

Glu Val Lys Ser Lys Lys His Pro Gln Ile Ile Lys Lys Gly Thr Ser

850 855 860

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

865 870 875 880

Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp

885 890 895

Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp

900 905 910

Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp

915 920 925

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

930 935 940

Asp Arg His Arg Ile Glu Glu Lys Arg Lys Arg Thr Tyr Glu Thr Phe

945 950 955 960

Lys Ser Ile Met Lys Lys Ser Pro Phe Ser Gly Pro Thr Asp Pro Arg

965 970 975

Pro Pro Pro Arg Arg Ile Ala Val Pro Ser Arg Ser Ser Ala Ser Val

980 985 990

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

995 1000 1005

Ile Asn Tyr Asp Glu Phe Pro Thr Met Val Phe Pro Ser Gly Gln

1010 1015 1020

Ile Ser Gln Ala Ser Ala Leu Ala Pro Ala Pro Pro Gln Val Leu

1025 1030 1035

Pro Gln Ala Pro Ala Pro Ala Pro Ala Pro Ala Met Val Ser Ala

1040 1045 1050

Leu Ala Gln Ala Pro Ala Pro Val Pro Val Leu Ala Pro Gly Pro

1055 1060 1065

Pro Gln Ala Val Ala Pro Pro Ala Pro Lys Pro Thr Gln Ala Gly

1070 1075 1080

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

1085 1090 1095

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

1100 1105 1110

Phe Thr Asp Leu Ala Ser Val Asp Asn Ser Glu Phe Gln Gln Leu

1115 1120 1125

Leu Asn Gln Gly Ile Pro Val Ala Pro His Thr Thr Glu Pro Met

1130 1135 1140

Leu Met Glu Tyr Pro Glu Ala Ile Thr Arg Leu Val Thr Gly Ala

1145 1150 1155

Gln Arg Pro Pro Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly

1160 1165 1170

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

1175 1180 1185

Ala Asp Met Asp Phe Ser Ala Leu Leu Gly Ser Gly Ser Gly Ser

1190 1195 1200

Arg Asp Ser Arg Glu Gly Met Phe Leu Pro Lys Pro Glu Ala Gly

1205 1210 1215

Ser Ala Ile Ser Asp Val Phe Glu Gly Arg Glu Val Cys Gln Pro

1220 1225 1230

Lys Arg Ile Arg Pro Phe His Pro Pro Gly Ser Pro Trp Ala Asn

1235 1240 1245

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

1250 1255 1260

His Glu Pro Val Gly Ser Leu Thr Pro Ala Pro Val Pro Gln Pro

1265 1270 1275

Leu Asp Pro Ala Pro Ala Val Thr Pro Glu Ala Ser His Leu Leu

1280 1285 1290

Glu Asp Pro Asp Glu Glu Thr Ser Gln Ala Val Lys Ala Leu Arg

1295 1300 1305

Glu Met Ala Asp Thr Val Ile Pro Gln Lys Glu Glu Ala Ala Ile

1310 1315 1320

Cys Gly Gln Met Asp Leu Ser His Pro Pro Pro Arg Gly His Leu

1325 1330 1335

Asp Glu Leu Thr Thr Thr Leu Glu Ser Met Thr Glu Asp Leu Asn

1340 1345 1350

Leu Asp Ser Pro Leu Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr

1355 1360 1365

Phe Leu Asn Asp Glu Cys Leu Leu His Ala Met His Ile Ser Thr

1370 1375 1380

Gly Leu Ser Ile Phe Asp Thr Ser Leu Phe

1385 1390

<210> 14

<211> 1225

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 14

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Lys Arg

1 5 10 15

Asn Tyr Ile Leu Gly Leu Asp Ile Gly Ile Thr Ser Val Gly Tyr Gly

20 25 30

Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly Val Arg Leu

35 40 45

Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg Ser Lys Arg

50 55 60

Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile Gln Arg Val

65 70 75 80

Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His Ser Glu Leu

85 90 95

Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu Ser Gln Lys

100 105 110

Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu Ala Lys Arg

115 120 125

Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr Gly Asn Glu

130 135 140

Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala Leu Glu Glu

145 150 155 160

Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys Asp Gly Glu

165 170 175

Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr Val Lys Glu

180 185 190

Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln Leu Asp Gln

195 200 205

Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg Arg Thr Tyr

210 215 220

Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys Asp Ile Lys

225 230 235 240

Glu Trp Tyr Glu Met Leu Lys Arg Arg Arg Arg His Arg Ile Leu Ser

245 250 255

Pro Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala

260 265 270

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

275 280 285

Ser Lys Arg Tyr Ala Thr Arg Gly Leu Met Asn Leu Leu Arg Ser Tyr

290 295 300

Phe Arg Val Asn Asn Leu Asp Val Lys Val Lys Ser Ile Asn Gly Gly

305 310 315 320

Phe Thr Ser Phe Leu Arg Arg Lys Trp Lys Phe Lys Lys Glu Arg Asn

325 330 335

Lys Gly Tyr Lys His His Ala Glu Asp Ala Leu Ile Ile Ala Asn Ala

340 345 350

Asp Phe Ile Phe Lys Glu Trp Lys Lys Leu Asp Lys Ala Lys Lys Val

355 360 365

Met Glu Asn Gln Met Phe Glu Glu Lys Gln Ala Glu Ser Met Pro Glu

370 375 380

Ile Glu Thr Glu Gln Glu Tyr Lys Glu Ile Phe Ile Thr Pro His Gln

385 390 395 400

Ile Lys His Ile Lys Asp Phe Lys Asp Tyr Lys Tyr Ser His Arg Val

405 410 415

Asp Lys Lys Pro Asn Arg Lys Leu Ile Asn Asp Thr Leu Tyr Ser Thr

420 425 430

Arg Lys Asp Asp Lys Gly Asn Thr Leu Ile Val Asn Asn Leu Asn Gly

435 440 445

Leu Tyr Asp Lys Asp Asn Asp Lys Leu Lys Lys Leu Ile Asn Lys Ser

450 455 460

Pro Glu Lys Leu Leu Met Tyr His His Asp Pro Gln Thr Tyr Gln Lys

465 470 475 480

Leu Lys Leu Ile Met Glu Gln Tyr Gly Asp Glu Lys Asn Pro Leu Tyr

485 490 495

Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr Leu Thr Lys Tyr Ser Lys Lys

500 505 510

Asp Asn Gly Pro Val Ile Lys Lys Ile Lys Tyr Tyr Gly Asn Lys Leu

515 520 525

Asn Ala His Leu Asp Ile Thr Asp Asp Tyr Pro Asn Ser Arg Asn Lys

530 535 540

Val Val Lys Leu Ser Leu Lys Pro Tyr Arg Phe Asp Val Tyr Leu Asp

545 550 555 560

Asn Gly Val Tyr Lys Phe Val Thr Val Lys Asn Leu Asp Val Ile Lys

565 570 575

Lys Glu Asn Tyr Tyr Glu Val Asn Ser Lys Cys Tyr Glu Glu Ala Lys

580 585 590

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

595 600 605

Lys Asn Asp Leu Ile Lys Ile Asn Gly Glu Leu Tyr Arg Val Ile Gly

610 615 620

Val Asn Asn Asp Leu Leu Asn Arg Ile Glu Val Asn Met Ile Asp Ile

625 630 635 640

Thr Tyr Arg Glu Tyr Leu Glu Asn Met Asn Asp Lys Arg Pro Pro His

645 650 655

Ile Ile Lys Thr Ile Ala Ser Lys Thr Gln Ser Ile Lys Lys Tyr Ser

660 665 670

Thr Asp Ile Leu Gly Asn Leu Tyr Glu Val Lys Ser Lys Lys His Pro

675 680 685

Gln Ile Ile Lys Lys Gly Thr Ser Arg Ala Asp Pro Lys Lys Lys Arg

690 695 700

Lys Val Glu Ala Ser Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe

705 710 715 720

Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp

725 730 735

Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly

740 745 750

Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg

755 760 765

Ser Gln Tyr Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys

770 775 780

Arg Lys Arg Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro

785 790 795 800

Phe Ser Gly Pro Thr Asp Pro Arg Pro Pro Pro Arg Arg Ile Ala Val

805 810 815

Pro Ser Arg Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln Pro Tyr

820 825 830

Pro Phe Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu Phe Pro Thr

835 840 845

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

850 855 860

Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala Pro Ala Pro Ala Pro

865 870 875 880

Ala Met Val Ser Ala Leu Ala Gln Ala Pro Ala Pro Val Pro Val Leu

885 890 895

Ala Pro Gly Pro Pro Gln Ala Val Ala Pro Pro Ala Pro Lys Pro Thr

900 905 910

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

915 920 925

Asp Asp Glu Asp Leu Gly Ala Leu Leu Gly Asn Ser Thr Asp Pro Ala

930 935 940

Val Phe Thr Asp Leu Ala Ser Val Asp Asn Ser Glu Phe Gln Gln Leu

945 950 955 960

Leu Asn Gln Gly Ile Pro Val Ala Pro His Thr Thr Glu Pro Met Leu

965 970 975

Met Glu Tyr Pro Glu Ala Ile Thr Arg Leu Val Thr Gly Ala Gln Arg

980 985 990

Pro Pro Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn

995 1000 1005

Gly Leu Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met

1010 1015 1020

Asp Phe Ser Ala Leu Leu Gly Ser Gly Ser Gly Ser Arg Asp Ser

1025 1030 1035

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

1040 1045 1050

Ser Asp Val Phe Glu Gly Arg Glu Val Cys Gln Pro Lys Arg Ile

1055 1060 1065

Arg Pro Phe His Pro Pro Gly Ser Pro Trp Ala Asn Arg Pro Leu

1070 1075 1080

Pro Ala Ser Leu Ala Pro Thr Pro Thr Gly Pro Val His Glu Pro

1085 1090 1095

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

1100 1105 1110

Ala Pro Ala Val Thr Pro Glu Ala Ser His Leu Leu Glu Asp Pro

1115 1120 1125

Asp Glu Glu Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met Ala

1130 1135 1140

Asp Thr Val Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln

1145 1150 1155

Met Asp Leu Ser His Pro Pro Pro Arg Gly His Leu Asp Glu Leu

1160 1165 1170

Thr Thr Thr Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp Ser

1175 1180 1185

Pro Leu Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn

1190 1195 1200

Asp Glu Cys Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser

1205 1210 1215

Ile Phe Asp Thr Ser Leu Phe

1220 1225

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<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 15

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Met Thr

1 5 10 15

Gln Phe Glu Gly Phe Thr Asn Leu Tyr Gln Val Ser Lys Thr Leu Arg

20 25 30

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

35 40 45

Gly Phe Ile Glu Glu Asp Lys Ala Arg Asn Asp His Tyr Lys Glu Leu

50 55 60

Lys Pro Ile Ile Asp Arg Ile Tyr Lys Thr Tyr Ala Asp Gln Cys Leu

65 70 75 80

Gln Leu Val Gln Leu Asp Trp Glu Asn Leu Ser Ala Ala Ile Asp Ser

85 90 95

Tyr Arg Lys Glu Lys Thr Glu Glu Thr Arg Asn Ala Leu Ile Glu Glu

100 105 110

Gln Ala Thr Tyr Arg Asn Ala Ile His Asp Tyr Phe Ile Gly Arg Thr

115 120 125

Asp Asn Leu Thr Asp Ala Ile Asn Lys Arg His Ala Glu Ile Tyr Lys

130 135 140

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

145 150 155 160

Gly Thr Val Thr Thr Thr Glu His Glu Asn Ala Leu Leu Arg Ser Phe

165 170 175

Asp Lys Phe Thr Thr Tyr Phe Ser Gly Phe Tyr Glu Asn Arg Lys Asn

180 185 190

Val Phe Ser Ala Glu Asp Ile Ser Thr Ala Ile Pro His Arg Ile Val

195 200 205

Gln Asp Asn Phe Pro Lys Phe Lys Glu Asn Cys His Ile Phe Thr Arg

210 215 220

Leu Ile Thr Ala Val Pro Ser Leu Arg Glu His Phe Glu Asn Val Lys

225 230 235 240

Lys Ala Ile Gly Ile Phe Val Ser Thr Ser Ile Glu Glu Val Phe Ser

245 250 255

Phe Pro Phe Tyr Asn Gln Leu Leu Thr Gln Thr Gln Ile Asp Leu Tyr

260 265 270

Asn Gln Leu Leu Gly Gly Ile Ser Arg Glu Ala Gly Thr Glu Lys Ile

275 280 285

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

290 295 300

Thr Ala His Ile Ile Ala Ser Leu Pro His Arg Phe Ile Pro Leu Phe

305 310 315 320

Lys Gln Ile Leu Ser Asp Arg Asn Thr Leu Ser Phe Ile Leu Glu Glu

325 330 335

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

340 345 350

Leu Leu Arg Asn Glu Asn Val Leu Glu Thr Ala Glu Ala Leu Phe Asn

355 360 365

Glu Leu Asn Ser Ile Asp Leu Thr His Ile Phe Ile Ser His Lys Lys

370 375 380

Leu Glu Thr Ile Ser Ser Ala Leu Cys Asp His Trp Asp Thr Leu Arg

385 390 395 400

Asn Ala Leu Tyr Glu Arg Arg Ile Ser Glu Leu Thr Gly Lys Ile Thr

405 410 415

Lys Ser Ala Lys Glu Lys Val Gln Arg Ser Leu Lys His Glu Asp Ile

420 425 430

Asn Leu Gln Glu Ile Ile Ser Ala Ala Gly Lys Glu Leu Ser Glu Ala

435 440 445

Phe Lys Gln Lys Thr Ser Glu Ile Leu Ser His Ala His Ala Ala Leu

450 455 460

Asp Gln Pro Leu Pro Thr Thr Leu Lys Lys Gln Glu Glu Lys Glu Ile

465 470 475 480

Leu Lys Ser Gln Leu Asp Ser Leu Leu Gly Leu Tyr His Leu Leu Asp

485 490 495

Trp Phe Ala Val Asp Glu Ser Asn Glu Val Asp Pro Glu Phe Ser Ala

500 505 510

Arg Leu Thr Gly Ile Lys Leu Glu Met Glu Pro Ser Leu Ser Phe Tyr

515 520 525

Asn Lys Ala Arg Asn Tyr Ala Thr Lys Lys Pro Tyr Ser Val Glu Lys

530 535 540

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

545 550 555 560

Asn Lys Glu Lys Asn Asn Gly Ala Ile Leu Phe Val Lys Asn Gly Leu

565 570 575

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

580 585 590

Ser Phe Glu Pro Thr Glu Lys Thr Ser Glu Gly Phe Asp Lys Met Tyr

595 600 605

Tyr Asp Tyr Phe Pro Asp Ala Ala Lys Met Ile Pro Lys Cys Ser Thr

610 615 620

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

625 630 635 640

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

645 650 655

Tyr Asp Leu Asn Asn Pro Glu Lys Glu Pro Lys Lys Phe Gln Thr Ala

660 665 670

Tyr Ala Lys Lys Thr Gly Asp Gln Lys Gly Tyr Arg Glu Ala Leu Cys

675 680 685

Lys Trp Ile Asp Phe Thr Arg Asp Phe Leu Ser Lys Tyr Thr Lys Thr

690 695 700

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

705 710 715 720

Leu Gly Glu Tyr Tyr Ala Glu Leu Asn Pro Leu Leu Tyr His Ile Ser

725 730 735

Phe Gln Arg Ile Ala Glu Lys Glu Ile Met Asp Ala Val Glu Thr Gly

740 745 750

Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ala Lys Gly His

755 760 765

His Gly Lys Pro Asn Leu His Thr Leu Tyr Trp Thr Gly Leu Phe Ser

770 775 780

Pro Glu Asn Leu Ala Lys Thr Ser Ile Lys Leu Asn Gly Gln Ala Glu

785 790 795 800

Leu Phe Tyr Arg Pro Lys Ser Arg Met Lys Arg Met Ala His Arg Leu

805 810 815

Gly Glu Lys Met Leu Asn Lys Lys Leu Lys Asp Gln Lys Thr Pro Ile

820 825 830

Pro Asp Thr Leu Tyr Gln Glu Leu Tyr Asp Tyr Val Asn His Arg Leu

835 840 845

Ser His Asp Leu Ser Asp Glu Ala Arg Ala Leu Leu Pro Asn Val Ile

850 855 860

Thr Lys Glu Val Ser His Glu Ile Ile Lys Asp Arg Arg Phe Thr Ser

865 870 875 880

Asp Lys Phe Phe Phe His Val Pro Ile Thr Leu Asn Tyr Gln Ala Ala

885 890 895

Asn Ser Pro Ser Lys Phe Asn Gln Arg Val Asn Ala Tyr Leu Lys Glu

900 905 910

His Pro Glu Thr Pro Ile Ile Gly Ile Ala Arg Gly Glu Arg Asn Leu

915 920 925

Ile Tyr Ile Thr Val Ile Asp Ser Thr Gly Lys Ile Leu Glu Gln Arg

930 935 940

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

945 950 955 960

Arg Glu Lys Glu Arg Val Ala Ala Arg Gln Ala Trp Ser Val Val Gly

965 970 975

Thr Ile Lys Asp Leu Lys Gln Gly Tyr Leu Ser Gln Val Ile His Glu

980 985 990

Ile Val Asp Leu Met Ile His Tyr Gln Ala Val Val Val Leu Glu Asn

995 1000 1005

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

1010 1015 1020

Ala Val Tyr Gln Gln Phe Glu Lys Met Leu Ile Asp Lys Leu Asn

1025 1030 1035

Cys Leu Val Leu Lys Asp Tyr Pro Ala Glu Lys Val Gly Gly Val

1040 1045 1050

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

1055 1060 1065

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

1070 1075 1080

Thr Ser Lys Ile Asp Pro Leu Thr Gly Phe Val Asp Pro Phe Val

1085 1090 1095

Trp Lys Thr Ile Lys Asn His Glu Ser Arg Lys His Phe Leu Glu

1100 1105 1110

Gly Phe Asp Phe Leu His Tyr Asp Val Lys Thr Gly Asp Phe Ile

1115 1120 1125

Leu His Phe Lys Met Asn Arg Asn Leu Ser Phe Gln Arg Gly Leu

1130 1135 1140

Pro Gly Phe Met Pro Ala Trp Asp Ile Val Phe Glu Lys Asn Glu

1145 1150 1155

Thr Gln Phe Asp Ala Lys Gly Thr Pro Phe Ile Ala Gly Lys Arg

1160 1165 1170

Ile Val Pro Val Ile Glu Asn His Arg Phe Thr Gly Arg Tyr Arg

1175 1180 1185

Asp Leu Tyr Pro Ala Asn Glu Leu Ile Ala Leu Leu Glu Glu Lys

1190 1195 1200

Gly Ile Val Phe Arg Asp Gly Ser Asn Ile Leu Pro Lys Leu Leu

1205 1210 1215

Glu Asn Asp Asp Ser His Ala Ile Asp Thr Met Val Ala Leu Ile

1220 1225 1230

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

1235 1240 1245

Asp Tyr Ile Asn Ser Pro Val Arg Asp Leu Asn Gly Val Cys Phe

1250 1255 1260

Asp Ser Arg Phe Gln Asn Pro Glu Trp Pro Met Asp Ala Asp Ala

1265 1270 1275

Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Gln Leu Leu Leu Asn

1280 1285 1290

His Leu Lys Glu Ser Lys Asp Leu Lys Leu Gln Asn Gly Ile Ser

1295 1300 1305

Asn Gln Asp Trp Leu Ala Tyr Ile Gln Glu Leu Arg Asn Thr Ser

1310 1315 1320

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

1325 1330 1335

Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly

1340 1345 1350

Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp

1355 1360 1365

Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu

1370 1375 1380

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

1385 1390 1395

Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg Lys

1400 1405 1410

Arg Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro Phe

1415 1420 1425

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

1430 1435 1440

Pro Ser Arg Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln Pro

1445 1450 1455

Tyr Pro Phe Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu Phe

1460 1465 1470

Pro Thr Met Val Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser Ala

1475 1480 1485

Leu Ala Pro Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala Pro

1490 1495 1500

Ala Pro Ala Pro Ala Met Val Ser Ala Leu Ala Gln Ala Pro Ala

1505 1510 1515

Pro Val Pro Val Leu Ala Pro Gly Pro Pro Gln Ala Val Ala Pro

1520 1525 1530

Pro Ala Pro Lys Pro Thr Gln Ala Gly Glu Gly Thr Leu Ser Glu

1535 1540 1545

Ala Leu Leu Gln Leu Gln Phe Asp Asp Glu Asp Leu Gly Ala Leu

1550 1555 1560

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

1565 1570 1575

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

1580 1585 1590

Val Ala Pro His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro Glu

1595 1600 1605

Ala Ile Thr Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp Pro

1610 1615 1620

Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu Leu

1625 1630 1635

Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe Ser

1640 1645 1650

Ala Leu Leu Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu Gly

1655 1660 1665

Met Phe Leu Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp Val

1670 1675 1680

Phe Glu Gly Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro Phe

1685 1690 1695

His Pro Pro Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala Ser

1700 1705 1710

Leu Ala Pro Thr Pro Thr Gly Pro Val His Glu Pro Val Gly Ser

1715 1720 1725

Leu Thr Pro Ala Pro Val Pro Gln Pro Leu Asp Pro Ala Pro Ala

1730 1735 1740

Val Thr Pro Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu Glu

1745 1750 1755

Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr Val

1760 1765 1770

Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp Leu

1775 1780 1785

Ser His Pro Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr Thr

1790 1795 1800

Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu Thr

1805 1810 1815

Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu Cys

1820 1825 1830

Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe Asp

1835 1840 1845

Thr Ser Leu Phe

1850

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<220>

<223> 通过合理设计产生的合成构建体

<400> 16

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Met Thr

1 5 10 15

Gln Phe Glu Gly Phe Thr Asn Leu Tyr Gln Val Ser Lys Thr Leu Arg

20 25 30

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

35 40 45

Gly Phe Ile Glu Glu Asp Lys Ala Arg Asn Asp His Tyr Lys Glu Leu

50 55 60

Lys Pro Ile Ile Asp Arg Ile Tyr Lys Thr Tyr Ala Asp Gln Cys Leu

65 70 75 80

Gln Leu Val Gln Leu Asp Trp Glu Asn Leu Ser Ala Ala Ile Asp Ser

85 90 95

Tyr Arg Lys Glu Lys Thr Glu Glu Thr Arg Asn Ala Leu Ile Glu Glu

100 105 110

Gln Ala Thr Tyr Arg Asn Ala Ile His Asp Tyr Phe Ile Gly Arg Thr

115 120 125

Asp Asn Leu Thr Asp Ala Ile Asn Lys Arg His Ala Glu Ile Tyr Lys

130 135 140

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

145 150 155 160

Gly Thr Val Thr Thr Thr Glu His Glu Asn Ala Leu Leu Arg Ser Phe

165 170 175

Asp Lys Phe Thr Thr Tyr Phe Ser Gly Phe Tyr Glu Asn Arg Lys Asn

180 185 190

Val Phe Ser Ala Glu Asp Ile Ser Thr Ala Ile Pro His Arg Ile Val

195 200 205

Gln Asp Asn Phe Pro Lys Phe Lys Glu Asn Cys His Ile Phe Thr Arg

210 215 220

Leu Ile Thr Ala Val Pro Ser Leu Arg Glu His Phe Glu Asn Val Lys

225 230 235 240

Lys Ala Ile Gly Ile Phe Val Ser Thr Ser Ile Glu Glu Val Phe Ser

245 250 255

Phe Pro Phe Tyr Asn Gln Leu Leu Thr Gln Thr Gln Ile Asp Leu Tyr

260 265 270

Asn Gln Leu Leu Gly Gly Ile Ser Arg Glu Ala Gly Thr Glu Lys Ile

275 280 285

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

290 295 300

Thr Ala His Ile Ile Ala Ser Leu Pro His Arg Phe Ile Pro Leu Phe

305 310 315 320

Lys Gln Ile Leu Ser Asp Arg Asn Thr Leu Ser Phe Ile Leu Glu Glu

325 330 335

Phe Gly Gly Gly Ser Tyr Ser Val Glu Lys Phe Lys Leu Asn Phe Gln

340 345 350

Met Pro Thr Leu Ala Ser Gly Trp Asp Val Asn Lys Glu Lys Asn Asn

355 360 365

Gly Ala Ile Leu Phe Val Lys Asn Gly Leu Tyr Tyr Leu Gly Ile Met

370 375 380

Pro Lys Gln Lys Gly Arg Tyr Lys Ala Leu Ser Phe Glu Pro Thr Glu

385 390 395 400

Lys Thr Ser Glu Gly Phe Asp Lys Met Tyr Tyr Asp Tyr Phe Pro Asp

405 410 415

Ala Ala Lys Met Ile Pro Lys Cys Ser Thr Gln Leu Lys Ala Val Thr

420 425 430

Ala His Phe Gln Thr His Thr Thr Pro Ile Leu Leu Ser Asn Asn Phe

435 440 445

Ile Glu Pro Leu Glu Ile Thr Lys Glu Ile Tyr Asp Leu Asn Asn Pro

450 455 460

Glu Lys Glu Pro Lys Lys Phe Gln Thr Ala Tyr Ala Lys Lys Thr Gly

465 470 475 480

Asp Gln Lys Gly Tyr Arg Glu Ala Leu Cys Lys Trp Ile Asp Phe Thr

485 490 495

Arg Asp Phe Leu Ser Lys Tyr Thr Lys Thr Thr Ser Ile Asp Leu Ser

500 505 510

Ser Leu Arg Pro Ser Ser Gln Tyr Lys Asp Leu Gly Glu Tyr Tyr Ala

515 520 525

Glu Leu Asn Pro Leu Leu Tyr His Ile Ser Phe Gln Arg Ile Ala Glu

530 535 540

Lys Glu Ile Met Asp Ala Val Glu Thr Gly Lys Leu Tyr Leu Phe Gln

545 550 555 560

Ile Tyr Asn Lys Asp Phe Ala Lys Gly His His Gly Lys Pro Asn Leu

565 570 575

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

580 585 590

Thr Ser Ile Lys Leu Asn Gly Gln Ala Glu Leu Phe Tyr Arg Pro Lys

595 600 605

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

610 615 620

Lys Lys Leu Lys Asp Gln Lys Thr Pro Ile Pro Asp Thr Leu Tyr Gln

625 630 635 640

Glu Leu Tyr Asp Tyr Val Asn His Arg Leu Ser His Asp Leu Ser Asp

645 650 655

Glu Ala Arg Ala Leu Leu Pro Asn Val Ile Thr Lys Glu Val Ser His

660 665 670

Glu Ile Ile Lys Asp Arg Arg Phe Thr Ser Asp Lys Phe Phe Phe His

675 680 685

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

690 695 700

Asn Gln Arg Val Asn Ala Tyr Leu Lys Glu His Pro Glu Thr Pro Ile

705 710 715 720

Ile Gly Ile Asp Arg Gly Glu Arg Asn Leu Ile Tyr Ile Thr Val Ile

725 730 735

Asp Ser Thr Gly Lys Ile Leu Glu Gln Arg Ser Leu Asn Thr Ile Gln

740 745 750

Gln Phe Asp Tyr Gln Lys Lys Leu Asp Asn Arg Glu Lys Glu Arg Val

755 760 765

Ala Ala Arg Gln Ala Trp Ser Val Val Gly Thr Ile Lys Asp Leu Lys

770 775 780

Gln Gly Tyr Leu Ser Gln Val Ile His Glu Ile Val Asp Leu Met Ile

785 790 795 800

His Tyr Gln Ala Val Val Val Leu Glu Asn Leu Asn Phe Gly Phe Lys

805 810 815

Ser Lys Arg Thr Gly Ile Ala Glu Lys Ala Val Tyr Gln Gln Phe Glu

820 825 830

Lys Met Leu Ile Asp Lys Leu Asn Cys Leu Val Leu Lys Asp Tyr Pro

835 840 845

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

850 855 860

Phe Thr Ser Phe Ala Lys Met Gly Thr Gln Ser Gly Phe Leu Phe Tyr

865 870 875 880

Val Pro Ala Pro Tyr Thr Ser Lys Ile Asp Pro Leu Thr Gly Phe Val

885 890 895

Asp Pro Phe Val Trp Lys Thr Ile Lys Asn His Glu Ser Arg Lys His

900 905 910

Phe Leu Glu Gly Phe Asp Phe Leu His Tyr Asp Val Lys Thr Gly Asp

915 920 925

Phe Ile Leu His Phe Lys Met Asn Arg Asn Leu Ser Phe Gln Arg Gly

930 935 940

Leu Pro Gly Phe Met Pro Ala Trp Asp Ile Val Phe Glu Lys Asn Glu

945 950 955 960

Thr Gln Phe Asp Ala Lys Gly Thr Pro Phe Ile Ala Gly Lys Arg Ile

965 970 975

Val Pro Val Ile Glu Asn His Arg Phe Thr Gly Arg Tyr Arg Asp Leu

980 985 990

Tyr Pro Ala Asn Glu Leu Ile Ala Leu Leu Glu Glu Lys Gly Ile Val

995 1000 1005

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

1010 1015 1020

Asp Ser His Ala Ile Asp Thr Met Val Ala Leu Ile Arg Ser Val

1025 1030 1035

Leu Gln Met Arg Asn Ser Asn Ala Ala Thr Gly Glu Asp Tyr Ile

1040 1045 1050

Asn Ser Pro Val Arg Asp Leu Asn Gly Val Cys Phe Asp Ser Arg

1055 1060 1065

Phe Gln Asn Pro Glu Trp Pro Met Asp Ala Asp Ala Asn Gly Ala

1070 1075 1080

Tyr His Ile Ala Leu Lys Gly Gln Leu Leu Leu Asn His Leu Lys

1085 1090 1095

Glu Ser Lys Asp Leu Lys Leu Gln Asn Gly Ile Ser Asn Gln Asp

1100 1105 1110

Trp Leu Ala Tyr Ile Gln Glu Leu Arg Asn Thr Ser Arg Ala Asp

1115 1120 1125

Pro Lys Lys Lys Arg Lys Val Glu Ala Ser Gly Ser Gly Arg Ala

1130 1135 1140

Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala

1145 1150 1155

Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp

1160 1165 1170

Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe

1175 1180 1185

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

1190 1195 1200

Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg Lys Arg Thr Tyr

1205 1210 1215

Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro Phe Ser Gly Pro

1220 1225 1230

Thr Asp Pro Arg Pro Pro Pro Arg Arg Ile Ala Val Pro Ser Arg

1235 1240 1245

Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln Pro Tyr Pro Phe

1250 1255 1260

Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu Phe Pro Thr Met

1265 1270 1275

Val Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser Ala Leu Ala Pro

1280 1285 1290

Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala Pro Ala Pro Ala

1295 1300 1305

Pro Ala Met Val Ser Ala Leu Ala Gln Ala Pro Ala Pro Val Pro

1310 1315 1320

Val Leu Ala Pro Gly Pro Pro Gln Ala Val Ala Pro Pro Ala Pro

1325 1330 1335

Lys Pro Thr Gln Ala Gly Glu Gly Thr Leu Ser Glu Ala Leu Leu

1340 1345 1350

Gln Leu Gln Phe Asp Asp Glu Asp Leu Gly Ala Leu Leu Gly Asn

1355 1360 1365

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

1370 1375 1380

Ser Glu Phe Gln Gln Leu Leu Asn Gln Gly Ile Pro Val Ala Pro

1385 1390 1395

His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro Glu Ala Ile Thr

1400 1405 1410

Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp Pro Ala Pro Ala

1415 1420 1425

Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu Leu Ser Gly Asp

1430 1435 1440

Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe Ser Ala Leu Leu

1445 1450 1455

Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu Gly Met Phe Leu

1460 1465 1470

Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp Val Phe Glu Gly

1475 1480 1485

Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro Phe His Pro Pro

1490 1495 1500

Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala Ser Leu Ala Pro

1505 1510 1515

Thr Pro Thr Gly Pro Val His Glu Pro Val Gly Ser Leu Thr Pro

1520 1525 1530

Ala Pro Val Pro Gln Pro Leu Asp Pro Ala Pro Ala Val Thr Pro

1535 1540 1545

Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu Glu Thr Ser Gln

1550 1555 1560

Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr Val Ile Pro Gln

1565 1570 1575

Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp Leu Ser His Pro

1580 1585 1590

Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr Thr Leu Glu Ser

1595 1600 1605

Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu Thr Pro Glu Leu

1610 1615 1620

Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu Cys Leu Leu His

1625 1630 1635

Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe Asp Thr Ser Leu

1640 1645 1650

Phe

<210> 17

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<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 17

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Lys Arg

1 5 10 15

Asn Tyr Ile Leu Gly Leu Ala Ile Gly Ile Thr Ser Val Gly Tyr Gly

20 25 30

Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly Val Arg Leu

35 40 45

Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg Ser Lys Arg

50 55 60

Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile Gln Arg Val

65 70 75 80

Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His Ser Glu Leu

85 90 95

Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu Ser Gln Lys

100 105 110

Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu Ala Lys Arg

115 120 125

Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr Gly Asn Glu

130 135 140

Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala Leu Glu Glu

145 150 155 160

Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys Asp Gly Glu

165 170 175

Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr Val Lys Glu

180 185 190

Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln Leu Asp Gln

195 200 205

Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg Arg Thr Tyr

210 215 220

Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys Asp Ile Lys

225 230 235 240

Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe Pro Glu Glu

245 250 255

Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr Asn Ala Leu

260 265 270

Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn Glu Lys Leu

275 280 285

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

290 295 300

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

305 310 315 320

Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys Pro Glu Phe

325 330 335

Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr Ala Arg Lys

340 345 350

Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala Lys Ile Leu

355 360 365

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

370 375 380

Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser Asn Leu Lys

385 390 395 400

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

405 410 415

Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala Ile Phe Asn

420 425 430

Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln Gln Lys Glu

435 440 445

Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro Val Val Lys

450 455 460

Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile Ile Lys Lys

465 470 475 480

Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg Glu Lys Asn

485 490 495

Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys Arg Asn Arg

500 505 510

Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr Gly Lys Glu

515 520 525

Asn Ala Lys Tyr Leu Ile Glu Lys Ile Lys Leu His Asp Met Gln Glu

530 535 540

Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu Asp Leu Leu

545 550 555 560

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

565 570 575

Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys Gln Glu Glu

580 585 590

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

595 600 605

Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile Leu Asn Leu

610 615 620

Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu Tyr Leu Leu

625 630 635 640

Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp Phe Ile Asn

645 650 655

Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu Met Asn Leu

660 665 670

Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys Val Lys Ser

675 680 685

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

690 695 700

Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp Ala Leu Ile

705 710 715 720

Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys Leu Asp Lys

725 730 735

Ala Lys Lys Val Met Glu Asn Gln Met Phe Gly Gly Gly Ser Gly Gly

740 745 750

Gly Ser Gly Gly Gly Ser Leu Val Lys Ser Glu Leu Glu Glu Lys Lys

755 760 765

Ser Glu Leu Arg His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu

770 775 780

Leu Ile Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met

785 790 795 800

Lys Val Met Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His

805 810 815

Leu Gly Gly Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser

820 825 830

Pro Ile Asp Tyr Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly

835 840 845

Tyr Asn Leu Pro Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu

850 855 860

Glu Asn Gln Thr Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys

865 870 875 880

Val Tyr Pro Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly

885 890 895

His Phe Lys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile

900 905 910

Thr Asn Cys Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly

915 920 925

Gly Glu Met Ile Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg

930 935 940

Lys Phe Asn Asn Gly Glu Ile Asn Phe Thr Ser Gly Gly Gly Ser Lys

945 950 955 960

Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys Ser

965 970 975

Arg

<210> 18

<211> 354

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 18

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Ala Glu

1 5 10 15

Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu Ile Phe Ile

20 25 30

Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp Tyr Lys Tyr

35 40 45

Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile Asn Asp Thr

50 55 60

Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu Ile Val Asn

65 70 75 80

Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu Lys Lys Leu

85 90 95

Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His Asp Pro Gln

100 105 110

Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly Asp Glu Lys

115 120 125

Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr Leu Thr Lys

130 135 140

Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile Lys Tyr Tyr

145 150 155 160

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

165 170 175

Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr Arg Phe Asp

180 185 190

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

195 200 205

Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser Lys Cys Tyr

210 215 220

Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala Glu Phe Ile

225 230 235 240

Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly Glu Leu Tyr

245 250 255

Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg Ile Glu Val Asn

260 265 270

Met Ile Asp Ile Thr Tyr Arg Glu Tyr Leu Glu Asn Met Asn Asp Lys

275 280 285

Arg Pro Pro His Ile Ile Lys Thr Ile Ala Ser Lys Thr Gln Ser Ile

290 295 300

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

305 310 315 320

Lys Lys His Pro Gln Ile Ile Lys Lys Gly Thr Ser Gly Gly Gly Ser

325 330 335

Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys

340 345 350

Ser Arg

<210> 19

<211> 809

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 19

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Lys Arg

1 5 10 15

Asn Tyr Ile Leu Gly Leu Ala Ile Gly Ile Thr Ser Val Gly Tyr Gly

20 25 30

Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly Val Arg Leu

35 40 45

Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg Ser Lys Arg

50 55 60

Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile Gln Arg Val

65 70 75 80

Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His Ser Glu Leu

85 90 95

Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu Ser Gln Lys

100 105 110

Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu Ala Lys Arg

115 120 125

Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr Gly Asn Glu

130 135 140

Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala Leu Glu Glu

145 150 155 160

Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys Asp Gly Glu

165 170 175

Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr Val Lys Glu

180 185 190

Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln Leu Asp Gln

195 200 205

Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg Arg Thr Tyr

210 215 220

Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys Asp Ile Lys

225 230 235 240

Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe Pro Glu Glu

245 250 255

Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr Asn Ala Leu

260 265 270

Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn Glu Lys Leu

275 280 285

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

290 295 300

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

305 310 315 320

Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys Pro Glu Phe

325 330 335

Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr Ala Arg Lys

340 345 350

Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala Lys Ile Leu

355 360 365

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

370 375 380

Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser Asn Leu Lys

385 390 395 400

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

405 410 415

Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala Ile Phe Asn

420 425 430

Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln Gln Lys Glu

435 440 445

Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro Val Val Lys

450 455 460

Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile Ile Lys Lys

465 470 475 480

Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Gly Ser Lys Arg Tyr

485 490 495

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

500 505 510

Asn Leu Asp Val Lys Val Lys Ser Ile Asn Gly Gly Phe Thr Ser Phe

515 520 525

Leu Arg Arg Lys Trp Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys

530 535 540

His His Ala Glu Asp Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe

545 550 555 560

Lys Glu Trp Lys Lys Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln

565 570 575

Met Phe Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Leu Val

580 585 590

Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu Lys

595 600 605

Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn Ser

610 615 620

Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met Lys

625 630 635 640

Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro Asp

645 650 655

Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile Val

660 665 670

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

675 680 685

Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys His

690 695 700

Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr Glu

705 710 715 720

Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys Ala

725 730 735

Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val Leu

740 745 750

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

755 760 765

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

770 775 780

Phe Thr Ser Gly Gly Gly Ser Lys Arg Pro Ala Ala Thr Lys Lys Ala

785 790 795 800

Gly Gln Ala Lys Lys Lys Lys Ser Arg

805

<210> 20

<211> 1409

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 20

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Met Ser

1 5 10 15

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

20 25 30

Glu Pro His Glu Phe Glu Val Phe Phe Asp Pro Arg Glu Leu Arg Lys

35 40 45

Glu Thr Cys Leu Leu Tyr Glu Ile Asn Trp Gly Gly Arg His Ser Ile

50 55 60

Trp Arg His Thr Ser Gln Asn Thr Asn Lys His Val Glu Val Asn Phe

65 70 75 80

Ile Glu Lys Phe Thr Thr Glu Arg Tyr Phe Cys Pro Asn Thr Arg Cys

85 90 95

Ser Ile Thr Trp Phe Leu Ser Trp Ser Pro Cys Gly Glu Cys Ser Arg

100 105 110

Ala Ile Thr Glu Phe Leu Ser Arg Tyr Pro His Val Thr Leu Phe Ile

115 120 125

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

130 135 140

Leu Arg Asp Leu Ile Ser Ser Gly Val Thr Ile Gln Ile Met Thr Glu

145 150 155 160

Gln Glu Ser Gly Tyr Cys Trp Arg Asn Phe Val Asn Tyr Ser Pro Ser

165 170 175

Asn Glu Ala His Trp Pro Arg Tyr Pro His Leu Trp Val Arg Leu Tyr

180 185 190

Val Leu Glu Leu Tyr Cys Ile Ile Leu Gly Leu Pro Pro Cys Leu Asn

195 200 205

Ile Leu Arg Arg Lys Gln Pro Gln Leu Thr Phe Phe Thr Ile Ala Leu

210 215 220

Gln Ser Cys His Tyr Gln Arg Leu Pro Pro His Ile Leu Trp Ala Thr

225 230 235 240

Gly Leu Lys Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr

245 250 255

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

260 265 270

Val Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala

275 280 285

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

290 295 300

Arg Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg

305 310 315 320

Ile Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp

325 330 335

His Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly

340 345 350

Leu Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His

355 360 365

Leu Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp

370 375 380

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

385 390 395 400

Ala Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys

405 410 415

Lys Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp

420 425 430

Tyr Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His

435 440 445

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

450 455 460

Arg Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp

465 470 475 480

Lys Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr

485 490 495

Phe Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu

500 505 510

Tyr Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu

515 520 525

Asn Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val

530 535 540

Phe Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile

545 550 555 560

Leu Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly

565 570 575

Lys Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile

580 585 590

Thr Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile

595 600 605

Ala Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu

610 615 620

Leu Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile

625 630 635 640

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

645 650 655

Ile Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile

660 665 670

Ala Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser

675 680 685

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

690 695 700

Pro Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala

705 710 715 720

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

725 730 735

Arg Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln

740 745 750

Lys Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr

755 760 765

Thr Gly Lys Glu Asn Ala Lys Tyr Leu Ile Glu Lys Ile Lys Leu His

770 775 780

Asp Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu

785 790 795 800

Glu Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile

805 810 815

Pro Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val

820 825 830

Lys Gln Glu Glu Ala Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr

835 840 845

Leu Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His

850 855 860

Ile Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys

865 870 875 880

Glu Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys

885 890 895

Asp Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly

900 905 910

Leu Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val

915 920 925

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

930 935 940

Trp Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu

945 950 955 960

Asp Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys

965 970 975

Lys Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu

980 985 990

Lys Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys

995 1000 1005

Glu Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe

1010 1015 1020

Lys Asp Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg

1025 1030 1035

Lys Leu Ile Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys

1040 1045 1050

Gly Asn Thr Leu Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys

1055 1060 1065

Asp Asn Asp Lys Leu Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys

1070 1075 1080

Leu Leu Met Tyr His His Asp Pro Gln Thr Tyr Gln Lys Leu Lys

1085 1090 1095

Leu Ile Met Glu Gln Tyr Gly Asp Glu Lys Asn Pro Leu Tyr Lys

1100 1105 1110

Tyr Tyr Glu Glu Thr Gly Asn Tyr Leu Thr Lys Tyr Ser Lys Lys

1115 1120 1125

Asp Asn Gly Pro Val Ile Lys Lys Ile Lys Tyr Tyr Gly Asn Lys

1130 1135 1140

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

1145 1150 1155

Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr Arg Phe Asp Val

1160 1165 1170

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

1175 1180 1185

Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser Lys Cys

1190 1195 1200

Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala Glu

1205 1210 1215

Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly

1220 1225 1230

Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg

1235 1240 1245

Ile Glu Val Asn Met Ile Asp Ile Thr Tyr Arg Glu Tyr Leu Glu

1250 1255 1260

Asn Met Asn Asp Lys Arg Pro Pro His Ile Ile Lys Thr Ile Ala

1265 1270 1275

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

1280 1285 1290

Asn Leu Tyr Glu Val Lys Ser Lys Lys His Pro Gln Ile Ile Lys

1295 1300 1305

Lys Gly Ser Gly Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys

1310 1315 1320

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

1325 1330 1335

Pro Glu Glu Val Glu Glu Val Ile Gly Asn Lys Pro Glu Ser Asp

1340 1345 1350

Ile Leu Val His Thr Ala Tyr Asp Glu Ser Thr Asp Glu Asn Val

1355 1360 1365

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

1370 1375 1380

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

1385 1390 1395

Gly Gly Ser Pro Pro Lys Lys Lys Arg Lys Val

1400 1405

<210> 21

<211> 1241

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 21

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Met Ser

1 5 10 15

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

20 25 30

Glu Pro His Glu Phe Glu Val Phe Phe Asp Pro Arg Glu Leu Arg Lys

35 40 45

Glu Thr Cys Leu Leu Tyr Glu Ile Asn Trp Gly Gly Arg His Ser Ile

50 55 60

Trp Arg His Thr Ser Gln Asn Thr Asn Lys His Val Glu Val Asn Phe

65 70 75 80

Ile Glu Lys Phe Thr Thr Glu Arg Tyr Phe Cys Pro Asn Thr Arg Cys

85 90 95

Ser Ile Thr Trp Phe Leu Ser Trp Ser Pro Cys Gly Glu Cys Ser Arg

100 105 110

Ala Ile Thr Glu Phe Leu Ser Arg Tyr Pro His Val Thr Leu Phe Ile

115 120 125

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

130 135 140

Leu Arg Asp Leu Ile Ser Ser Gly Val Thr Ile Gln Ile Met Thr Glu

145 150 155 160

Gln Glu Ser Gly Tyr Cys Trp Arg Asn Phe Val Asn Tyr Ser Pro Ser

165 170 175

Asn Glu Ala His Trp Pro Arg Tyr Pro His Leu Trp Val Arg Leu Tyr

180 185 190

Val Leu Glu Leu Tyr Cys Ile Ile Leu Gly Leu Pro Pro Cys Leu Asn

195 200 205

Ile Leu Arg Arg Lys Gln Pro Gln Leu Thr Phe Phe Thr Ile Ala Leu

210 215 220

Gln Ser Cys His Tyr Gln Arg Leu Pro Pro His Ile Leu Trp Ala Thr

225 230 235 240

Gly Leu Lys Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr

245 250 255

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

260 265 270

Val Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala

275 280 285

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

290 295 300

Arg Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg

305 310 315 320

Ile Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp

325 330 335

His Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly

340 345 350

Leu Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His

355 360 365

Leu Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp

370 375 380

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

385 390 395 400

Ala Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys

405 410 415

Lys Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp

420 425 430

Tyr Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His

435 440 445

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

450 455 460

Arg Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp

465 470 475 480

Lys Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr

485 490 495

Phe Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu

500 505 510

Tyr Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu

515 520 525

Asn Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val

530 535 540

Phe Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile

545 550 555 560

Leu Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly

565 570 575

Lys Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile

580 585 590

Thr Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile

595 600 605

Ala Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu

610 615 620

Leu Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile

625 630 635 640

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

645 650 655

Ile Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile

660 665 670

Ala Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser

675 680 685

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

690 695 700

Pro Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala

705 710 715 720

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

725 730 735

Ser Lys Arg Tyr Ala Thr Arg Gly Leu Met Asn Leu Leu Arg Ser Tyr

740 745 750

Phe Arg Val Asn Asn Leu Asp Val Lys Val Lys Ser Ile Asn Gly Gly

755 760 765

Phe Thr Ser Phe Leu Arg Arg Lys Trp Lys Phe Lys Lys Glu Arg Asn

770 775 780

Lys Gly Tyr Lys His His Ala Glu Asp Ala Leu Ile Ile Ala Asn Ala

785 790 795 800

Asp Phe Ile Phe Lys Glu Trp Lys Lys Leu Asp Lys Ala Lys Lys Val

805 810 815

Met Glu Asn Gln Met Phe Glu Glu Lys Gln Ala Glu Ser Met Pro Glu

820 825 830

Ile Glu Thr Glu Gln Glu Tyr Lys Glu Ile Phe Ile Thr Pro His Gln

835 840 845

Ile Lys His Ile Lys Asp Phe Lys Asp Tyr Lys Tyr Ser His Arg Val

850 855 860

Asp Lys Lys Pro Asn Arg Lys Leu Ile Asn Asp Thr Leu Tyr Ser Thr

865 870 875 880

Arg Lys Asp Asp Lys Gly Asn Thr Leu Ile Val Asn Asn Leu Asn Gly

885 890 895

Leu Tyr Asp Lys Asp Asn Asp Lys Leu Lys Lys Leu Ile Asn Lys Ser

900 905 910

Pro Glu Lys Leu Leu Met Tyr His His Asp Pro Gln Thr Tyr Gln Lys

915 920 925

Leu Lys Leu Ile Met Glu Gln Tyr Gly Asp Glu Lys Asn Pro Leu Tyr

930 935 940

Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr Leu Thr Lys Tyr Ser Lys Lys

945 950 955 960

Asp Asn Gly Pro Val Ile Lys Lys Ile Lys Tyr Tyr Gly Asn Lys Leu

965 970 975

Asn Ala His Leu Asp Ile Thr Asp Asp Tyr Pro Asn Ser Arg Asn Lys

980 985 990

Val Val Lys Leu Ser Leu Lys Pro Tyr Arg Phe Asp Val Tyr Leu Asp

995 1000 1005

Asn Gly Val Tyr Lys Phe Val Thr Val Lys Asn Leu Asp Val Ile

1010 1015 1020

Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser Lys Cys Tyr Glu Glu

1025 1030 1035

Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala Glu Phe Ile Ala

1040 1045 1050

Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly Glu Leu Tyr

1055 1060 1065

Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg Ile Glu Val

1070 1075 1080

Asn Met Ile Asp Ile Thr Tyr Arg Glu Tyr Leu Glu Asn Met Asn

1085 1090 1095

Asp Lys Arg Pro Pro His Ile Ile Lys Thr Ile Ala Ser Lys Thr

1100 1105 1110

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

1115 1120 1125

Glu Val Lys Ser Lys Lys His Pro Gln Ile Ile Lys Lys Gly Ser

1130 1135 1140

Gly Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly

1145 1150 1155

Lys Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu

1160 1165 1170

Val Glu Glu Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val

1175 1180 1185

His Thr Ala Tyr Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu

1190 1195 1200

Thr Ser Asp Ala Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln

1205 1210 1215

Asp Ser Asn Gly Glu Asn Lys Ile Lys Met Leu Ser Gly Gly Ser

1220 1225 1230

Pro Pro Lys Lys Lys Arg Lys Val

1235 1240

<210> 22

<211> 771

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 22

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Lys Arg

1 5 10 15

Asn Tyr Ile Leu Gly Leu Ala Ile Gly Ile Thr Ser Val Gly Tyr Gly

20 25 30

Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly Val Arg Leu

35 40 45

Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg Ser Lys Arg

50 55 60

Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile Gln Arg Val

65 70 75 80

Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His Ser Glu Leu

85 90 95

Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu Ser Gln Lys

100 105 110

Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu Ala Lys Arg

115 120 125

Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr Gly Asn Glu

130 135 140

Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala Leu Glu Glu

145 150 155 160

Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys Asp Gly Glu

165 170 175

Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr Val Lys Glu

180 185 190

Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln Leu Asp Gln

195 200 205

Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg Arg Thr Tyr

210 215 220

Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys Asp Ile Lys

225 230 235 240

Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe Pro Glu Glu

245 250 255

Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr Asn Ala Leu

260 265 270

Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn Glu Lys Leu

275 280 285

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

290 295 300

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

305 310 315 320

Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys Pro Glu Phe

325 330 335

Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr Ala Arg Lys

340 345 350

Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala Lys Ile Leu

355 360 365

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

370 375 380

Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser Asn Leu Lys

385 390 395 400

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

405 410 415

Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala Ile Phe Asn

420 425 430

Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln Gln Lys Glu

435 440 445

Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro Val Val Lys

450 455 460

Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile Ile Lys Lys

465 470 475 480

Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg Glu Lys Gly

485 490 495

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

500 505 510

Val Asp Pro Thr Leu Arg Arg Arg Ile Glu Pro His Glu Phe Glu Val

515 520 525

Phe Phe Asp Pro Arg Glu Leu Arg Lys Glu Thr Cys Leu Leu Tyr Glu

530 535 540

Ile Asn Trp Gly Gly Arg His Ser Ile Trp Arg His Thr Ser Gln Asn

545 550 555 560

Thr Asn Lys His Val Glu Val Asn Phe Ile Glu Lys Phe Thr Thr Glu

565 570 575

Arg Tyr Phe Cys Pro Asn Thr Arg Cys Ser Ile Thr Trp Phe Leu Ser

580 585 590

Trp Ser Pro Cys Gly Glu Cys Ser Arg Ala Ile Thr Glu Phe Leu Ser

595 600 605

Arg Tyr Pro His Val Thr Leu Phe Ile Tyr Ile Ala Arg Leu Tyr His

610 615 620

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

625 630 635 640

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

645 650 655

Arg Asn Phe Val Asn Tyr Ser Pro Ser Asn Glu Ala His Trp Pro Arg

660 665 670

Tyr Pro His Leu Trp Val Arg Leu Tyr Val Leu Glu Leu Tyr Cys Ile

675 680 685

Ile Leu Gly Leu Pro Pro Cys Leu Asn Ile Leu Arg Arg Lys Gln Pro

690 695 700

Gln Leu Thr Phe Phe Thr Ile Ala Leu Gln Ser Cys His Tyr Gln Arg

705 710 715 720

Leu Pro Pro His Ile Leu Trp Ala Thr Gly Leu Lys Ser Gly Ser Glu

725 730 735

Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Thr Ser Gly Gly Gly

740 745 750

Ser Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys

755 760 765

Lys Ser Arg

770

<210> 23

<211> 518

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<400> 23

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Thr Arg

1 5 10 15

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

20 25 30

Asn Asn Leu Asp Val Lys Val Lys Ser Ile Asn Gly Gly Phe Thr Ser

35 40 45

Phe Leu Arg Arg Lys Trp Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr

50 55 60

Lys His His Ala Glu Asp Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile

65 70 75 80

Phe Lys Glu Trp Lys Lys Leu Asp Lys Ala Lys Lys Val Met Glu Asn

85 90 95

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

100 105 110

Glu Gln Glu Tyr Lys Glu Ile Phe Ile Thr Pro His Gln Ile Lys His

115 120 125

Ile Lys Asp Phe Lys Asp Tyr Lys Tyr Ser His Arg Val Asp Lys Lys

130 135 140

Pro Asn Arg Lys Leu Ile Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp

145 150 155 160

Asp Lys Gly Asn Thr Leu Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp

165 170 175

Lys Asp Asn Asp Lys Leu Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys

180 185 190

Leu Leu Met Tyr His His Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu

195 200 205

Ile Met Glu Gln Tyr Gly Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr

210 215 220

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

225 230 235 240

Pro Val Ile Lys Lys Ile Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His

245 250 255

Leu Asp Ile Thr Asp Asp Tyr Pro Asn Ser Arg Asn Lys Val Val Lys

260 265 270

Leu Ser Leu Lys Pro Tyr Arg Phe Asp Val Tyr Leu Asp Asn Gly Val

275 280 285

Tyr Lys Phe Val Thr Val Lys Asn Leu Asp Val Ile Lys Lys Glu Asn

290 295 300

Tyr Tyr Glu Val Asn Ser Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys

305 310 315 320

Lys Ile Ser Asn Gln Ala Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp

325 330 335

Leu Ile Lys Ile Asn Gly Glu Leu Tyr Arg Val Ile Gly Val Asn Asn

340 345 350

Asp Leu Leu Asn Arg Ile Glu Val Asn Met Ile Asp Ile Thr Tyr Arg

355 360 365

Glu Tyr Leu Glu Asn Met Asn Asp Lys Arg Pro Pro His Ile Ile Lys

370 375 380

Thr Ile Ala Ser Lys Thr Gln Ser Ile Lys Lys Tyr Ser Thr Asp Ile

385 390 395 400

Leu Gly Asn Leu Tyr Glu Val Lys Ser Lys Lys His Pro Gln Ile Ile

405 410 415

Lys Lys Gly Ser Gly Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys

420 425 430

Glu Thr Gly Lys Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro

435 440 445

Glu Glu Val Glu Glu Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu

450 455 460

Val His Thr Ala Tyr Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu

465 470 475 480

Thr Ser Asp Ala Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp

485 490 495

Ser Asn Gly Glu Asn Lys Ile Lys Met Leu Ser Gly Gly Ser Pro Pro

500 505 510

Lys Lys Lys Arg Lys Val

515

<210> 24

<211> 391

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<223> 通过合理设计产生的合成构建体

<400> 24

Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu

1 5 10 15

Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp

20 25 30

Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp

35 40 45

Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Ala Leu Leu Gln Leu Gln

50 55 60

Phe Asp Asp Glu Asp Leu Gly Ala Leu Leu Gly Asn Ser Thr Asp Pro

65 70 75 80

Ala Val Phe Thr Asp Leu Ala Ser Val Asp Asn Ser Glu Phe Gln Gln

85 90 95

Leu Leu Asn Gln Gly Ile Pro Val Ala Pro His Thr Thr Glu Pro Met

100 105 110

Leu Met Glu Tyr Pro Glu Ala Ile Thr Arg Leu Val Thr Gly Ala Gln

115 120 125

Arg Pro Pro Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro

130 135 140

Asn Gly Leu Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met

145 150 155 160

Asp Phe Ser Ala Leu Leu Ser Gln Ile Ser Ser Gly Ser Gly Ser Gly

165 170 175

Ser Arg Asp Ser Arg Glu Gly Met Phe Leu Pro Lys Pro Glu Ala Gly

180 185 190

Ser Ala Ile Ser Asp Val Phe Glu Gly Arg Glu Val Cys Gln Pro Lys

195 200 205

Arg Ile Arg Pro Phe His Pro Pro Gly Ser Pro Trp Ala Asn Arg Pro

210 215 220

Leu Pro Ala Ser Leu Ala Pro Thr Pro Thr Gly Pro Val His Glu Pro

225 230 235 240

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

245 250 255

Pro Ala Val Thr Pro Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu

260 265 270

Glu Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr Val

275 280 285

Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp Leu Ser

290 295 300

His Pro Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr Thr Leu Glu

305 310 315 320

Ser Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu Thr Pro Glu Leu

325 330 335

Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu Cys Leu Leu His Ala

340 345 350

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

355 360 365

Ser Gly Gly Gly Ser Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln

370 375 380

Ala Lys Lys Lys Lys Ser Arg

385 390

<210> 25

<211> 382

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 25

Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu

1 5 10 15

Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp

20 25 30

Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp

35 40 45

Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg Leu Val Thr Gly Ala

50 55 60

Gln Arg Pro Pro Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu

65 70 75 80

Pro Asn Gly Leu Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp

85 90 95

Met Asp Phe Ser Ala Leu Leu Ser Gln Ile Ser Ser Gly Gly Gly Ser

100 105 110

Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp Pro Ala Pro Ala Pro

115 120 125

Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu Leu Ser Gly Asp Glu Asp

130 135 140

Phe Ser Ser Ile Ala Asp Met Asp Phe Ser Ala Leu Leu Ser Gln Ile

145 150 155 160

Ser Ser Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu Gly Met Phe

165 170 175

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

180 185 190

Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro Phe His Pro Pro Gly

195 200 205

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

210 215 220

Thr Gly Pro Val His Glu Pro Val Gly Ser Leu Thr Pro Ala Pro Val

225 230 235 240

Pro Gln Pro Leu Asp Pro Ala Pro Ala Val Thr Pro Glu Ala Ser His

245 250 255

Leu Leu Glu Asp Pro Asp Glu Glu Thr Ser Gln Ala Val Lys Ala Leu

260 265 270

Arg Glu Met Ala Asp Thr Val Ile Pro Gln Lys Glu Glu Ala Ala Ile

275 280 285

Cys Gly Gln Met Asp Leu Ser His Pro Pro Pro Arg Gly His Leu Asp

290 295 300

Glu Leu Thr Thr Thr Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp

305 310 315 320

Ser Pro Leu Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn

325 330 335

Asp Glu Cys Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser Ile

340 345 350

Phe Asp Thr Ser Leu Phe Thr Ser Gly Gly Gly Ser Lys Arg Pro Ala

355 360 365

Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys Ser Arg

370 375 380

<210> 26

<211> 663

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<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 26

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Gly Ala

1 5 10 15

Met Val Asp Thr Leu Ser Gly Leu Ser Ser Glu Gln Gly Gln Ser Gly

20 25 30

Asp Met Thr Ile Glu Glu Asp Ser Ala Thr His Ile Lys Phe Ser Lys

35 40 45

Arg Asp Glu Asp Gly Lys Glu Leu Ala Gly Ala Thr Met Glu Leu Arg

50 55 60

Asp Ser Ser Gly Lys Thr Ile Ser Thr Trp Ile Ser Asp Gly Gln Val

65 70 75 80

Lys Asp Phe Tyr Leu Tyr Pro Gly Lys Tyr Thr Phe Val Glu Thr Ala

85 90 95

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

100 105 110

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

115 120 125

His Ile Thr Gly Thr Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val

130 135 140

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

145 150 155 160

Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu

165 170 175

Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp

180 185 190

Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg Ser Gln

195 200 205

Tyr Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg Lys

210 215 220

Arg Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro Phe Ser

225 230 235 240

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

245 250 255

Arg Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln Pro Tyr Pro Phe

260 265 270

Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu Phe Pro Thr Met Val

275 280 285

Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser Ala Leu Ala Pro Ala Pro

290 295 300

Pro Gln Val Leu Pro Gln Ala Pro Ala Pro Ala Pro Ala Pro Ala Met

305 310 315 320

Val Ser Ala Leu Ala Gln Ala Pro Ala Pro Val Pro Val Leu Ala Pro

325 330 335

Gly Pro Pro Gln Ala Val Ala Pro Pro Ala Pro Lys Pro Thr Gln Ala

340 345 350

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

355 360 365

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

370 375 380

Thr Asp Leu Ala Ser Val Asp Asn Ser Glu Phe Gln Gln Leu Leu Asn

385 390 395 400

Gln Gly Ile Pro Val Ala Pro His Thr Thr Glu Pro Met Leu Met Glu

405 410 415

Tyr Pro Glu Ala Ile Thr Arg Leu Val Thr Gly Ala Gln Arg Pro Pro

420 425 430

Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu

435 440 445

Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe Ser

450 455 460

Ala Leu Leu Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu Gly Met

465 470 475 480

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

485 490 495

Gly Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro Phe His Pro Pro

500 505 510

Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala Ser Leu Ala Pro Thr

515 520 525

Pro Thr Gly Pro Val His Glu Pro Val Gly Ser Leu Thr Pro Ala Pro

530 535 540

Val Pro Gln Pro Leu Asp Pro Ala Pro Ala Val Thr Pro Glu Ala Ser

545 550 555 560

His Leu Leu Glu Asp Pro Asp Glu Glu Thr Ser Gln Ala Val Lys Ala

565 570 575

Leu Arg Glu Met Ala Asp Thr Val Ile Pro Gln Lys Glu Glu Ala Ala

580 585 590

Ile Cys Gly Gln Met Asp Leu Ser His Pro Pro Pro Arg Gly His Leu

595 600 605

Asp Glu Leu Thr Thr Thr Leu Glu Ser Met Thr Glu Asp Leu Asn Leu

610 615 620

Asp Ser Pro Leu Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu

625 630 635 640

Asn Asp Glu Cys Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser

645 650 655

Ile Phe Asp Thr Ser Leu Phe

660

<210> 27

<211> 88

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 27

Gly Gly Gly Ser Gly Gly Ala His Ile Val Met Val Asp Ala Tyr Lys

1 5 10 15

Pro Thr Lys Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Ala His Ile

20 25 30

Val Met Val Asp Ala Tyr Lys Pro Thr Lys Gly Gly Ser Gly Gly Gly

35 40 45

Gly Ser Gly Gly Ala His Ile Val Met Val Asp Ala Tyr Lys Pro Thr

50 55 60

Lys Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Ala His Ile Val Met

65 70 75 80

Val Asp Ala Tyr Lys Pro Thr Lys

85

<210> 28

<211> 643

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 28

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Asn His

1 5 10 15

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

20 25 30

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

35 40 45

Arg Asp Ile Lys Gly Asn Glu Leu Ala Gly Ala Thr Ile Glu Leu Arg

50 55 60

Asp Leu Ser Gly Lys Ser Ile Gln Ser Trp Val Ser Asp Gly Lys Ala

65 70 75 80

Lys Asp Phe Tyr Leu Leu Pro Gly Ser Tyr Glu Phe Val Glu Thr Ala

85 90 95

Ala Pro Glu Gly Tyr Gln Ile Ala Thr Lys Ile Met Phe Thr Ile Ser

100 105 110

Thr Asp Gly Arg Ile Thr Val Asp Gly Gln Leu Val Thr Gly Thr Gly

115 120 125

Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly

130 135 140

Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala

145 150 155 160

Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp

165 170 175

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

180 185 190

Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg Lys Arg Thr Tyr Glu

195 200 205

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

210 215 220

Pro Arg Pro Pro Pro Arg Arg Ile Ala Val Pro Ser Arg Ser Ser Ala

225 230 235 240

Ser Val Pro Lys Pro Ala Pro Gln Pro Tyr Pro Phe Thr Ser Ser Leu

245 250 255

Ser Thr Ile Asn Tyr Asp Glu Phe Pro Thr Met Val Phe Pro Ser Gly

260 265 270

Gln Ile Ser Gln Ala Ser Ala Leu Ala Pro Ala Pro Pro Gln Val Leu

275 280 285

Pro Gln Ala Pro Ala Pro Ala Pro Ala Pro Ala Met Val Ser Ala Leu

290 295 300

Ala Gln Ala Pro Ala Pro Val Pro Val Leu Ala Pro Gly Pro Pro Gln

305 310 315 320

Ala Val Ala Pro Pro Ala Pro Lys Pro Thr Gln Ala Gly Glu Gly Thr

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

Val Ala Pro His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro Glu Ala

385 390 395 400

Ile Thr Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp Pro Ala Pro

405 410 415

Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu Leu Ser Gly Asp

420 425 430

Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe Ser Ala Leu Leu Gly

435 440 445

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

450 455 460

Pro Glu Ala Gly Ser Ala Ile Ser Asp Val Phe Glu Gly Arg Glu Val

465 470 475 480

Cys Gln Pro Lys Arg Ile Arg Pro Phe His Pro Pro Gly Ser Pro Trp

485 490 495

Ala Asn Arg Pro Leu Pro Ala Ser Leu Ala Pro Thr Pro Thr Gly Pro

500 505 510

Val His Glu Pro Val Gly Ser Leu Thr Pro Ala Pro Val Pro Gln Pro

515 520 525

Leu Asp Pro Ala Pro Ala Val Thr Pro Glu Ala Ser His Leu Leu Glu

530 535 540

Asp Pro Asp Glu Glu Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met

545 550 555 560

Ala Asp Thr Val Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln

565 570 575

Met Asp Leu Ser His Pro Pro Pro Arg Gly His Leu Asp Glu Leu Thr

580 585 590

Thr Thr Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu

595 600 605

Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu Cys

610 615 620

Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe Asp Thr

625 630 635 640

Ser Leu Phe

<210> 29

<211> 78

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 29

Gly Gly Gly Ser Gly Gly Ala His Ile Val Met Val Asp Asn Tyr Lys

1 5 10 15

Pro Ile Val Gly Gly Ser Gly Gly Ala His Ile Val Met Val Asp Asn

20 25 30

Tyr Lys Pro Ile Val Gly Gly Ser Gly Gly Ala His Ile Val Met Val

35 40 45

Asp Asn Tyr Lys Pro Ile Val Gly Gly Ser Gly Gly Gly Gly Ser Gly

50 55 60

Gly Ala His Ile Val Met Val Asp Asn Tyr Lys Pro Ile Val

65 70 75

<210> 30

<211> 285

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 30

Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu

1 5 10 15

Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp

20 25 30

Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp

35 40 45

Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg Leu Val Thr Gly Ala

50 55 60

Gln Arg Pro Pro Asp Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu

65 70 75 80

Pro Asn Gly Leu Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp

85 90 95

Met Asp Phe Ser Ala Leu Leu Ser Gln Ile Ser Ser Gly Gly Gly Ser

100 105 110

Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp Pro Ala Pro Ala Pro

115 120 125

Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu Leu Ser Gly Asp Glu Asp

130 135 140

Phe Ser Ser Ile Ala Asp Met Asp Phe Ser Ala Leu Leu Ser Gln Ile

145 150 155 160

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

165 170 175

Pro Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu Glu Thr Ser Gln

180 185 190

Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr Val Ile Pro Gln Lys

195 200 205

Glu Glu Ala Ala Ile Cys Gly Gln Met Asp Leu Ser His Pro Pro Pro

210 215 220

Arg Gly His Leu Asp Glu Leu Thr Thr Thr Leu Glu Ser Met Thr Glu

225 230 235 240

Asp Leu Asn Leu Asp Ser Pro Leu Thr Pro Glu Leu Asn Glu Ile Leu

245 250 255

Asp Thr Phe Leu Asn Asp Glu Cys Leu Leu His Ala Met His Ile Ser

260 265 270

Thr Gly Leu Ser Ile Phe Asp Thr Ser Leu Phe Thr Ser

275 280 285

<210> 31

<211> 20

<212> DNA

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 31

gctgtgtttg cgtctctccc 20

<210> 32

<211> 20

<212> DNA

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 32

ggggtggtga caagtgtgat 20

<210> 33

<211> 1433

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 33

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Gly Gly

1 5 10 15

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

20 25 30

Val Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala

35 40 45

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

50 55 60

Arg Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg

65 70 75 80

Ile Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp

85 90 95

His Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly

100 105 110

Leu Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His

115 120 125

Leu Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp

130 135 140

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

145 150 155 160

Ala Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys

165 170 175

Lys Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp

180 185 190

Tyr Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His

195 200 205

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

210 215 220

Arg Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp

225 230 235 240

Lys Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr

245 250 255

Phe Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu

260 265 270

Tyr Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu

275 280 285

Asn Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val

290 295 300

Phe Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile

305 310 315 320

Leu Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly

325 330 335

Lys Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile

340 345 350

Thr Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile

355 360 365

Ala Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu

370 375 380

Leu Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile

385 390 395 400

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

405 410 415

Ile Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile

420 425 430

Ala Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser

435 440 445

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

450 455 460

Pro Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala

465 470 475 480

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

485 490 495

Ser Lys Arg Tyr Ala Thr Arg Gly Leu Met Asn Leu Leu Arg Ser Tyr

500 505 510

Phe Arg Val Asn Asn Leu Asp Val Lys Val Lys Ser Ile Asn Gly Gly

515 520 525

Phe Thr Ser Phe Leu Arg Arg Lys Trp Lys Phe Lys Lys Glu Arg Asn

530 535 540

Lys Gly Tyr Lys His His Ala Glu Asp Ala Leu Ile Ile Ala Asn Ala

545 550 555 560

Asp Phe Ile Phe Lys Glu Trp Lys Lys Leu Asp Lys Ala Lys Lys Val

565 570 575

Met Glu Asn Gln Met Phe Glu Glu Lys Gln Ala Glu Ser Met Pro Glu

580 585 590

Ile Glu Thr Glu Gln Glu Tyr Lys Glu Ile Phe Ile Thr Pro His Gln

595 600 605

Ile Lys His Ile Lys Asp Phe Lys Asp Tyr Lys Tyr Ser His Arg Val

610 615 620

Asp Lys Lys Pro Asn Arg Lys Leu Ile Asn Asp Thr Leu Tyr Ser Thr

625 630 635 640

Arg Lys Asp Asp Lys Gly Asn Thr Leu Ile Val Asn Asn Leu Asn Gly

645 650 655

Leu Tyr Asp Lys Asp Asn Asp Lys Leu Lys Lys Leu Ile Asn Lys Ser

660 665 670

Pro Glu Lys Leu Leu Met Tyr His His Asp Pro Gln Thr Tyr Gln Lys

675 680 685

Leu Lys Leu Ile Met Glu Gln Tyr Gly Asp Glu Lys Asn Pro Leu Tyr

690 695 700

Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr Leu Thr Lys Tyr Ser Lys Lys

705 710 715 720

Asp Asn Gly Pro Val Ile Lys Lys Ile Lys Tyr Tyr Gly Asn Lys Leu

725 730 735

Asn Ala His Leu Asp Ile Thr Asp Asp Tyr Pro Asn Ser Arg Asn Lys

740 745 750

Val Val Lys Leu Ser Leu Lys Pro Tyr Arg Phe Asp Val Tyr Leu Asp

755 760 765

Asn Gly Val Tyr Lys Phe Val Thr Val Lys Asn Leu Asp Val Ile Lys

770 775 780

Lys Glu Asn Tyr Tyr Glu Val Asn Ser Lys Cys Tyr Glu Glu Ala Lys

785 790 795 800

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

805 810 815

Lys Asn Asp Leu Ile Lys Ile Asn Gly Glu Leu Tyr Arg Val Ile Gly

820 825 830

Val Asn Asn Asp Leu Leu Asn Arg Ile Glu Val Asn Met Ile Asp Ile

835 840 845

Thr Tyr Arg Glu Tyr Leu Glu Asn Met Asn Asp Lys Arg Pro Pro His

850 855 860

Ile Ile Lys Thr Ile Ala Ser Lys Thr Gln Ser Ile Lys Lys Tyr Ser

865 870 875 880

Thr Asp Ile Leu Gly Asn Leu Tyr Glu Val Lys Ser Lys Lys His Pro

885 890 895

Gln Ile Ile Lys Lys Gly Thr Ser Arg Ala Asp Pro Lys Lys Lys Arg

900 905 910

Lys Val Glu Ala Ser Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe

915 920 925

Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp

930 935 940

Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly

945 950 955 960

Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg

965 970 975

Ser Gln Tyr Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys

980 985 990

Arg Lys Arg Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro

995 1000 1005

Phe Ser Gly Pro Thr Asp Pro Arg Pro Pro Pro Arg Arg Ile Ala

1010 1015 1020

Val Pro Ser Arg Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln

1025 1030 1035

Pro Tyr Pro Phe Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu

1040 1045 1050

Phe Pro Thr Met Val Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser

1055 1060 1065

Ala Leu Ala Pro Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala

1070 1075 1080

Pro Ala Pro Ala Pro Ala Met Val Ser Ala Leu Ala Gln Ala Pro

1085 1090 1095

Ala Pro Val Pro Val Leu Ala Pro Gly Pro Pro Gln Ala Val Ala

1100 1105 1110

Pro Pro Ala Pro Lys Pro Thr Gln Ala Gly Glu Gly Thr Leu Ser

1115 1120 1125

Glu Ala Leu Leu Gln Leu Gln Phe Asp Asp Glu Asp Leu Gly Ala

1130 1135 1140

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

1145 1150 1155

Ser Val Asp Asn Ser Glu Phe Gln Gln Leu Leu Asn Gln Gly Ile

1160 1165 1170

Pro Val Ala Pro His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro

1175 1180 1185

Glu Ala Ile Thr Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp

1190 1195 1200

Pro Ala Pro Ala Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu

1205 1210 1215

Leu Ser Gly Asp Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe

1220 1225 1230

Ser Ala Leu Leu Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu

1235 1240 1245

Gly Met Phe Leu Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp

1250 1255 1260

Val Phe Glu Gly Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro

1265 1270 1275

Phe His Pro Pro Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala

1280 1285 1290

Ser Leu Ala Pro Thr Pro Thr Gly Pro Val His Glu Pro Val Gly

1295 1300 1305

Ser Leu Thr Pro Ala Pro Val Pro Gln Pro Leu Asp Pro Ala Pro

1310 1315 1320

Ala Val Thr Pro Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu

1325 1330 1335

Glu Thr Ser Gln Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr

1340 1345 1350

Val Ile Pro Gln Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp

1355 1360 1365

Leu Ser His Pro Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr

1370 1375 1380

Thr Leu Glu Ser Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu

1385 1390 1395

Thr Pro Glu Leu Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu

1400 1405 1410

Cys Leu Leu His Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe

1415 1420 1425

Asp Thr Ser Leu Phe

1430

<210> 34

<211> 1414

<212> PRT

<213> 人工序列

<220>

<223> 通过合理设计产生的合成构建体

<400> 34

Met Pro Lys Lys Lys Arg Lys Val Gly Gly Gly Ser Pro Gly Gly Gly

1 5 10 15

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

20 25 30

Val Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala

35 40 45

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

50 55 60

Arg Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg

65 70 75 80

Ile Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp

85 90 95

His Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly

100 105 110

Leu Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His

115 120 125

Leu Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp

130 135 140

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

145 150 155 160

Ala Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys

165 170 175

Lys Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp

180 185 190

Tyr Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His

195 200 205

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

210 215 220

Arg Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp

225 230 235 240

Lys Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Lys

245 250 255

Lys Val Asp Leu Ser Gln Gln Lys Glu Ile Pro Thr Thr Leu Val Asp

260 265 270

Asp Phe Ile Leu Ser Pro Val Val Lys Arg Ser Phe Ile Gln Ser Ile

275 280 285

Lys Val Ile Asn Ala Ile Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile

290 295 300

Ile Ile Glu Leu Ala Arg Glu Lys Asn Ser Lys Asp Ala Gln Lys Met

305 310 315 320

Ile Asn Glu Met Gln Lys Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu

325 330 335

Glu Ile Ile Arg Thr Thr Gly Lys Glu Asn Ala Lys Tyr Leu Ile Glu

340 345 350

Lys Ile Lys Leu His Asp Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu

355 360 365

Glu Ala Ile Pro Leu Glu Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu

370 375 380

Val Asp His Ile Ile Pro Arg Ser Val Ser Phe Asp Asn Ser Phe Asn

385 390 395 400

Asn Lys Val Leu Val Lys Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg

405 410 415

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

420 425 430

Thr Phe Lys Lys His Ile Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile

435 440 445

Ser Lys Thr Lys Lys Glu Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg

450 455 460

Phe Ser Val Gln Lys Asp Phe Ile Asn Arg Asn Leu Val Asp Thr Arg

465 470 475 480

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

485 490 495

Asn Asn Leu Asp Val Lys Val Lys Ser Ile Asn Gly Gly Phe Thr Ser

500 505 510

Phe Leu Arg Arg Lys Trp Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr

515 520 525

Lys His His Ala Glu Asp Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile

530 535 540

Phe Lys Glu Trp Lys Lys Leu Asp Lys Ala Lys Lys Val Met Glu Asn

545 550 555 560

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

565 570 575

Glu Gln Glu Tyr Lys Glu Ile Phe Ile Thr Pro His Gln Ile Lys His

580 585 590

Ile Lys Asp Phe Lys Asp Tyr Lys Tyr Ser His Arg Val Asp Lys Lys

595 600 605

Pro Asn Arg Lys Leu Ile Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp

610 615 620

Asp Lys Gly Asn Thr Leu Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp

625 630 635 640

Lys Asp Asn Asp Lys Leu Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys

645 650 655

Leu Leu Met Tyr His His Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu

660 665 670

Ile Met Glu Gln Tyr Gly Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr

675 680 685

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

690 695 700

Pro Val Ile Lys Lys Ile Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His

705 710 715 720

Leu Asp Ile Thr Asp Asp Tyr Pro Asn Ser Arg Asn Lys Val Val Lys

725 730 735

Leu Ser Leu Lys Pro Tyr Arg Phe Asp Val Tyr Leu Asp Asn Gly Val

740 745 750

Tyr Lys Phe Val Thr Val Lys Asn Leu Asp Val Ile Lys Lys Glu Asn

755 760 765

Tyr Tyr Glu Val Asn Ser Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys

770 775 780

Lys Ile Ser Asn Gln Ala Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp

785 790 795 800

Leu Ile Lys Ile Asn Gly Glu Leu Tyr Arg Val Ile Gly Val Asn Asn

805 810 815

Asp Leu Leu Asn Arg Ile Glu Val Asn Met Ile Asp Ile Thr Tyr Arg

820 825 830

Glu Tyr Leu Glu Asn Met Asn Asp Lys Arg Pro Pro His Ile Ile Lys

835 840 845

Thr Ile Ala Ser Lys Thr Gln Ser Ile Lys Lys Tyr Ser Thr Asp Ile

850 855 860

Leu Gly Asn Leu Tyr Glu Val Lys Ser Lys Lys His Pro Gln Ile Ile

865 870 875 880

Lys Lys Gly Thr Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Glu

885 890 895

Ala Ser Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp

900 905 910

Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly

915 920 925

Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala

930 935 940

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

945 950 955 960

Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu Glu Lys Arg Lys Arg

965 970 975

Thr Tyr Glu Thr Phe Lys Ser Ile Met Lys Lys Ser Pro Phe Ser Gly

980 985 990

Pro Thr Asp Pro Arg Pro Pro Pro Arg Arg Ile Ala Val Pro Ser Arg

995 1000 1005

Ser Ser Ala Ser Val Pro Lys Pro Ala Pro Gln Pro Tyr Pro Phe

1010 1015 1020

Thr Ser Ser Leu Ser Thr Ile Asn Tyr Asp Glu Phe Pro Thr Met

1025 1030 1035

Val Phe Pro Ser Gly Gln Ile Ser Gln Ala Ser Ala Leu Ala Pro

1040 1045 1050

Ala Pro Pro Gln Val Leu Pro Gln Ala Pro Ala Pro Ala Pro Ala

1055 1060 1065

Pro Ala Met Val Ser Ala Leu Ala Gln Ala Pro Ala Pro Val Pro

1070 1075 1080

Val Leu Ala Pro Gly Pro Pro Gln Ala Val Ala Pro Pro Ala Pro

1085 1090 1095

Lys Pro Thr Gln Ala Gly Glu Gly Thr Leu Ser Glu Ala Leu Leu

1100 1105 1110

Gln Leu Gln Phe Asp Asp Glu Asp Leu Gly Ala Leu Leu Gly Asn

1115 1120 1125

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

1130 1135 1140

Ser Glu Phe Gln Gln Leu Leu Asn Gln Gly Ile Pro Val Ala Pro

1145 1150 1155

His Thr Thr Glu Pro Met Leu Met Glu Tyr Pro Glu Ala Ile Thr

1160 1165 1170

Arg Leu Val Thr Gly Ala Gln Arg Pro Pro Asp Pro Ala Pro Ala

1175 1180 1185

Pro Leu Gly Ala Pro Gly Leu Pro Asn Gly Leu Leu Ser Gly Asp

1190 1195 1200

Glu Asp Phe Ser Ser Ile Ala Asp Met Asp Phe Ser Ala Leu Leu

1205 1210 1215

Gly Ser Gly Ser Gly Ser Arg Asp Ser Arg Glu Gly Met Phe Leu

1220 1225 1230

Pro Lys Pro Glu Ala Gly Ser Ala Ile Ser Asp Val Phe Glu Gly

1235 1240 1245

Arg Glu Val Cys Gln Pro Lys Arg Ile Arg Pro Phe His Pro Pro

1250 1255 1260

Gly Ser Pro Trp Ala Asn Arg Pro Leu Pro Ala Ser Leu Ala Pro

1265 1270 1275

Thr Pro Thr Gly Pro Val His Glu Pro Val Gly Ser Leu Thr Pro

1280 1285 1290

Ala Pro Val Pro Gln Pro Leu Asp Pro Ala Pro Ala Val Thr Pro

1295 1300 1305

Glu Ala Ser His Leu Leu Glu Asp Pro Asp Glu Glu Thr Ser Gln

1310 1315 1320

Ala Val Lys Ala Leu Arg Glu Met Ala Asp Thr Val Ile Pro Gln

1325 1330 1335

Lys Glu Glu Ala Ala Ile Cys Gly Gln Met Asp Leu Ser His Pro

1340 1345 1350

Pro Pro Arg Gly His Leu Asp Glu Leu Thr Thr Thr Leu Glu Ser

1355 1360 1365

Met Thr Glu Asp Leu Asn Leu Asp Ser Pro Leu Thr Pro Glu Leu

1370 1375 1380

Asn Glu Ile Leu Asp Thr Phe Leu Asn Asp Glu Cys Leu Leu His

1385 1390 1395

Ala Met His Ile Ser Thr Gly Leu Ser Ile Phe Asp Thr Ser Leu

1400 1405 1410

Phe

Claims (20)

1.一种非天然存在的合理设计的缩减型Cas9，包含通过缺失保守功能结构域产生的保留DNA结合活性的尺寸缩小的金黄色葡萄球菌mini-SaCas9。

2.根据权利要求1所述的非天然存在的mini-SaCas9蛋白系统，其中所述mini-SaCas9的保守功能结构域在RuvC2区和RuvC3区之间被移除并且通过GSK接头连接。

3.根据权利要求2所述的非天然存在的mini-SaCas9蛋白系统，其中所述mini-SaCas9的保守功能结构域在所述RuvC2区和所述RuvC3区之间的氨基酸位置479-649处被移除并且通过所述GSK接头连接。

4.根据权利要求1所述的非天然存在的mini-SaCas9蛋白系统，其中所述mini-SaCas9的保守功能结构域在RuvC2区和RuvC3区之间被移除、通过GSK接头连接并且分割为两个或更多个部分。

5.根据权利要求4所述的非天然存在的mini-SaCas9蛋白系统，其中所述mini-SaCas9的保守功能结构域在所述RuvC2区和所述RuvC3区之间的残基位置479-649处被移除、通过所述GSK接头连接并且在残基738处分割为两个或更多个部分。

6.根据权利要求4所述的非天然存在的分割的mini-SaCas9蛋白系统，还包含跨不同分割对的分割Cas9部分，以产生即使当片段部分冗余时也提供完整多肽序列激活基因表达的组合。

7.根据权利要求3所述的非天然存在的mini-SaCas9蛋白系统，还包含FokI结构域。

8.根据权利要求5所述的非天然存在的分割的mini-SaCas9蛋白系统，还包含FokI结构域。

9.根据权利要求7所述的非天然存在的mini-SaCas9蛋白系统，其中所述miniSaCas9是mini-dSaCas9，并且还包含多个基因调节元件以在形成复合体时形成FokI二聚体构型。

10.根据权利要求9所述的非天然存在的mini-SaCas9蛋白系统，其中所述FokI二聚体复合体在二聚化的mini-dSaCas9之间产生4-40个碱基对的间隔。

11.根据权利要求9所述的非天然存在的mini-dSaCas9蛋白系统，其中所述FokI二聚体构型在二聚化的mini-dSaCas9之间产生14个碱基对的间隔。

12.根据权利要求8所述的非天然存在的分割的mini-SaCas9蛋白系统，其中所述分割的miniSaCas9是还包含多个基因调节元件以在形成复合体时形成二聚体构型的分割的mini-dSaCas9。

13.根据权利要求12所述的非天然存在的分割的mini-dSaCas9蛋白系统，其中所述FokI二聚体构型在二聚化的mini-dSaCas9之间产生12-24个碱基对的间隔。

14.根据权利要求12所述的非天然存在的mini-SaCas9蛋白系统，其中所述FokI二聚体构型在处于二聚体构型时恢复核酸酶活性。

15.根据权利要求2所述的非天然存在的合理设计的尺寸缩小的金黄色葡萄球菌mini-SaCas9，还包含重复肽Moontag阵列，所述重复肽Moontag阵列具有融合至mini-SaCas9-3的C-末端的13个残基MoonTag的四个串联重复序列，并且使MoonCatcher与VPR结构域形成复合体。

16.根据权利要求15所述的具有重复肽Moontag阵列的非天然存在的合理设计的尺寸缩小的金黄色葡萄球菌mini-SaCas9，其中所述Moontag对应于序列ID号29，并且MoonCatcher VPR对应于序列ID号28。

17.一种非天然存在的合理设计的指导RNA，包含经优化的指导RNA，其中四个重复U-A区通过在第三位置的A-U密码子翻转和在第四位置的G-C替换进行优化。

18.根据权利要求17所述的非天然存在的合理设计的指导RNA，还包含偶联至所述经优化的指导RNA的Glu tRNA。

19.一种非天然存在的合理设计的缩减型Cas9，包含通过缺失REC区和HNH区之间的保守功能结构域产生的保留DNA结合活性的尺寸缩小的金黄色葡萄球菌mini-SaCas9，并且被连接以将效应子结构域、gRNA表达盒和其它部分合并在单个AAV载体中以用于转录激活、DNA切割和碱基编辑。

20.根据权利要求19所述的非天然存在的合理设计的缩减型mini-SaCas9，其中所述gRNA表达盒在四个重复U-A区处通过在第三位置的A-U密码子翻转和第四位置的G-C替换进行优化并且还合并了与经优化的指导RNA偶联的Glu tRNA。

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