CN103998609A - 通过递送人工转录因子调节受体表达 - Google Patents
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Abstract
本发明涉及人工转录因子，其包含融合至抑制或激活蛋白结构域、核定位序列和蛋白质转导结构域的特异性靶向受体基因启动子的多指锌指蛋白。在具体的实例中，这些受体基因启动子调节内皮素受体A、内皮素受体B、Toll样受体4或高亲和力IgE受体的表达。指向内皮素A或B受体的人工转录因子用于治疗由内皮素调节的疾病，诸如心血管疾病和尤其是眼疾病，例如视网膜静脉阻塞、视网膜动脉阻塞、黄斑水肿、视神经病变、中心性浆液性脉络膜视网膜病变、视网膜色素变性、Leber遗传性视神经病变等。指向Toll样受体4或IgE受体的人工转录因子分别用于治疗自身免疫病症等，和变应性病症。
Description
发明领域
本发明涉及人工转录因子，其包含融合至抑制或激活结构域、核定位序列和蛋白质转导结构域的特异性靶向受体基因启动子的多指锌指蛋白，以及它们在治疗由特异性效应物与此类受体的结合所调节的疾病中的用途。
背景技术
提出人工转录因子（ATF）是用于调节基因表达的有用工具(Sera T., 2009, Adv Drug Deliv Rev 61, 513-526)。许多天然存在的转录因子通过抑制或激活基因转录来影响表达，具有识别某一DNA序列的复杂的特定结构域。如果技术人员意在修饰它们的特异性和一个或多个靶基因，则这使得它们对于操作而言是无吸引力的目标。然而，某一类转录因子含有几个所谓锌指（ZF）结构域，它们是模块化的，并因此使得它们可以进行遗传工程。锌指是几乎独立靶向三个DNA碱基对的短（30个氨基酸）DNA结合基序。含有几个此类锌指的蛋白因此能够识别更长的DNA序列。六聚锌指蛋白（ZFP）识别18个碱基对（bp）的DNA靶，其在整个人基因组中几乎是唯一的。最初认为是完全背景独立的，但更深入的分析显示对于锌指的某些背景特异性(Klug A., 2010, Annu Rev Biochem 79, 213-231)。突变在锌指识别表面中的某些氨基酸改变ZF模块的结合特异性产生对于大部分的5’-GNN-3’、5’-CNN-3’、5’-ANN-3’和某些5’-TNN-3’密码子的确定的ZF结构单元(例如所谓的Barbas模块，见Dreier B., Barbas C.F. 3rd 等, 2005, J Biol Chem 280, 35588-35597)。尽管关于人工转录因子的早期工作集中于基于组合预先选择的锌指与已知的3 bp靶序列的合理设计，但意识到锌指的某一背景特异性需要产生大的锌指文库，其使用复杂方法诸如细菌或酵母单杂交、噬菌体展示、区室化核糖体展示（compartmentalized ribosome display）或使用FACS分析的体内选择来询问。
使用此类人工锌指蛋白，可以以高特异性靶向在人基因组内的DNA基因座。因此，这些锌指蛋白是将具有转录调节活性的蛋白结构域转运至特定启动子序列以产生目标基因表达的调节的理想工具。用于转录沉默的合适结构域是Krueppel相关的结构域(KRAB)如N-末端(SEQ ID NO: 1)或C-末端(SEQ ID NO: 2) KRAB结构域、Sin3相互作用结构域(SID, SEQ ID NO: 3)和ERF阻抑蛋白结构域(ERD, SEQ ID NO: 4)，而基因转录的激活通过单纯疱疹病毒VP16 (SEQ ID NO: 5)或VP64 (VP16的四聚体重复, SEQ ID NO: 6)结构域来完成(Beerli R.R.等, 1998, Proc Natl Acad Sci USA 95, 14628-14633)。此外，考虑通过基因本体论GO:0001071 (http://amigo.geneontology.org/cgi-bin/amigo/term_details?term=GO:0001071)确定的蛋白的转录活性结构域实现靶蛋白的转录调节。
所有已知药物靶的很大百分比是受体分子，其被具有时常相当大的脱靶（off-target）活性的小分子药物的作用而刺激或阻断。此类受体的实例是组胺H1受体或α和β肾上腺素受体，但通常是通过基因本体论GO:0004888和GO:0004930确定的蛋白。
尽管由于特定特征的高保守性，小分子药物并非总能选择性靶向给定蛋白家族的某一成员，但如基于抗体的新药物所显示出的，生物制剂提供了巨大的特异性。然而，实际上所有生物制剂到目前为止均在细胞外起作用。
尤其是上文提及的人工转录因子将适合于以治疗上有用的方式影响基因转录。然而，此类因子向作用位点－细胞核－的递送并不容易实现，因而妨碍了治疗性人工转录因子方法的有效性，例如通过依赖于逆转录病毒递送具有该方法的所有缺点，诸如免疫原性和细胞转化的潜能(Lund C.V.等, 2005, Mol Cell Biol 25, 9082-9091)。
所谓的蛋白质转导结构域（PTD）显示促进蛋白经质膜易位至细胞溶胶/核质中。短肽诸如HIV来源的TAT肽(SEQ ID NO: 7)和其他显示出诱导不依赖于细胞类型的货物蛋白质的大胞饮摄取(Wadia J.S. et al., 2004, Nat Med 10, 310-315)。当到达细胞溶胶中时，此类融合蛋白显示出具有生物活性。有趣的是，在蛋白转导后，极可能是通过胞内蛋白伴侣的作用，即使错误折叠的蛋白也能够变得有功能。
血管活性内皮素系统在多种疾病的发病中起重要作用。内皮素一方面参与调节血液供给，并且另一方面是由低氧诱导的事件级联中的主要参与者。内皮素例如参与血脑或血视网膜屏障的破坏和参与新血管形成。此外，内皮素参与神经变性以及调节痛觉或甚至口渴感觉的阈值。内皮素还参与调节眼内压。
内皮素作用通过它的同族受体来介导，主要是内皮素受体A，其通常位于环绕血管的平滑肌细胞上。影响内皮素系统－全身地或局部地－是治疗许多疾病诸如蛛网膜下腔或脑出血所关注的。内皮素还影响多发性硬化的病程。内皮素是（肺动脉）高血压的原因，而且也是低动脉压、心肌病和雷诺综合征、变异型心绞痛和其他心血管疾病的原因。内皮素参与糖尿病性肾病和糖尿病性视网膜病变。在眼中，它还对青光眼性神经变性、视网膜静脉阻塞、巨细胞性关节炎、视网膜色素变性、年龄相关性黄斑变性、中心性浆液性脉络膜视网膜病变、Morbus Leber、Susac综合征、眼内出血、视网膜前神经胶质增生和某些其他病理学病况起作用。
眼是精密器官，其强烈依赖于平衡的且充足的灌注以达到它的高需氧量。无法提供充足且稳定的供氧会引起局部缺血-再灌注损伤，导致神经胶质活化和神经元损害，如在患有渐进性疾病的青光眼患者中所观察到的，尽管其具有正常或标准化的眼内压。如在糖尿病性视网膜病变或湿性年龄相关性黄斑变性中显而易见的，不充足的血液供给还导致低氧，引起具有进一步视网膜损害潜能的失控的（run-away）新血管形成。眼组织灌注在复杂控制之下并且依赖于血压、眼内压以及调节血管直径的局部因子。此类局部因子例如提及的内皮素、具有强血管收缩活性的短肽。内皮素的三种同工型(ET-1、ET-2和ET-3)由内皮素转化酶从位于血管壁中的内皮细胞分泌的前体分子来产生。成熟ET的两个同族受体是已知的，ETRA和ETRB。尽管ETRA位于形成血管壁的平滑肌细胞中并促进血管收缩，但ETRB主要表达于内皮细胞中，并且通过促进一氧化氮的释放来使血管舒张，因而引起平滑肌松弛。ETRA和ETRB属于G蛋白偶联的七跨膜螺旋受体的大类。ET与ETRA或ETRB的结合导致G蛋白活化，因而引发胞内钙浓度的升高，并由此引起一大批的细胞反应。
药理学上影响ET系统可能证明在其中ET水平升高并且ET以有害形式起作用的病例中，诸如在视网膜静脉阻塞、青光眼性神经变性、视网膜色素变性、巨细胞性关节炎、中心性浆液性脉络膜视网膜病变、多发性硬化、视神经炎、类风湿性关节炎、Susac综合征、辐射性视网膜病变、视网膜前神经胶质增生、纤维肌痛和糖尿病性视网膜病变过程中是有用的。为此目的，下调ETRA将有助于调节疾病结果。但在某些情况下，上调ETRA并因此对ET提高的敏感性可能是希望的，例如以促进在从角膜外伤或角膜溃疡恢复过程中的角膜伤口愈合。
此外，ETRB介导的信号传递例如在癌症干细胞维持和肿瘤生长过程中与病例生理学过程有关。此外，上调ETRB与青光眼性神经变性有关，而抑制ETRB则显示出在青光眼过程中起神经保护作用。此外，ETRB在炎症过程中被上调。因此，通过特异性人工转录因子药理学上调节ETRB将可用于治疗癌症，预防神经变性和调节炎症过程。
细菌细胞壁组分诸如脂多糖（LPS）在多种疾病的发病中起重要作用。体内存在LPS指向需要由免疫系统针对的细菌感染。由于LPS是革兰氏阴性细菌的一般组分，因此LPS构成了能够激活免疫系统的所谓的危险信号。LPS由Toll样受体4（TLR4）所识别，所述Toll样受体4是参与识别多种危险信号或与细菌或病毒感染相关的病原体相关分子模式（PAMP）的Toll样受体的更大家族的成员。尽管识别LPS为危险信号是先天免疫的重要部分，但TLR4受体的过度刺激或延长的刺激与多种与慢性炎症有关的病理学病况相关。实例是多种肝疾病，诸如酒精性肝病、非酒精性脂肪性肝病、非酒精性脂肪性肝炎、慢性乙型肝炎或丙型肝炎病毒（HCV）感染、和HIV-HCV共感染。其他与TLR4信号传递相关的疾病是类风湿性关节炎、动脉粥样硬化、银屑病、克罗恩氏病、葡萄膜炎、接触镜相关性角膜炎和角膜炎症。此外，TLR4介导的信号传递参与癌症进展（cancer progression）和对化学疗法的抗性。
药理学上影响LPS识别和TLR4信号传递可能证明对于与由于TLR4的不恰当活化而导致的慢性炎症有关的疾病是有用的。因此，通过靶向TLR4启动子的特异性负调节的人工转录因子的作用来下调TLR4蛋白，会有助于通过破坏由LPS导致的慢性炎症的恶性循环来调节疾病结果。
免疫球蛋白同种型E（IgE）是适应性免疫系统的部分，并且同样参与针对感染以及致瘤性转化的保护。IgE通过位于肥大细胞和嗜碱细胞上的高亲和力IgE受体（FCER1）来结合。IgE与FCER1的结合及随后经称为变应原的特异性抗原交联这些复合体导致多种因子从肥大细胞和嗜碱细胞中释放，引起变应性应答。在这些因子中有组胺、白细胞三烯类、多种细胞因子以及溶菌酶、类胰蛋白酶或β-己糖胺酶。这些因子的释放与变应性疾病诸如变应性鼻炎、哮喘、湿疹和过敏反应相关。
发明概述
本发明涉及人工转录因子，其包含融合至抑制或激活蛋白结构域、核定位序列和蛋白质转导结构域的特异性靶向受体基因启动子的多指锌指蛋白，并且涉及包含此类人工转录因子的药物组合物。此外，本发明涉及此类人工转录因子用于调节细胞对外界刺激和对其他可溶性信号分子的反应的用途，以及在治疗由特异性效应物与此类受体的结合所调节的疾病中的用途。
在一个具体的实施方案中，受体基因启动子是内皮素受体A启动子。在另一个具体的实施方案中，本发明涉及此类人工转录因子，其用于影响对内皮素的细胞应答，以降低或提高内皮素受体水平，并且用于治疗由内皮素调节的疾病，尤其用于治疗此类眼疾病。同样，本发明涉及治疗由内皮素调节的疾病的方法，其包括向有其需要的患者施用治疗有效量的本发明的人工转录因子。
在另一个具体的实施方案中，本发明涉及人工转录因子中间体，其包含融合至抑制或激活蛋白结构域和核定位序列的特异性靶向内皮素受体A启动子的多指锌指蛋白。
在另一个具体的实施方案中，受体基因启动子是内皮素受体B启动子。在另一个具体的实施方案中，本发明涉及此类人工转录因子，其用于影响对内皮素的细胞应答，以降低或提高内皮素受体B水平，并且用于治疗由内皮素调节的疾病，尤其用于治疗此类眼疾病。同样，本发明涉及治疗由内皮素调节的疾病的方法，其包括向有其需要的患者施用治疗有效量的本发明的人工转录因子。
在另一个具体的实施方案中，本发明涉及人工转录因子中间体，其包含融合至抑制或激活蛋白结构域和核定位序列的特异性靶向内皮素受体B启动子的多指锌指蛋白。
在另一个具体的实施方案中，受体基因启动子是Toll样受体4启动子。在另一个具体的实施方案中，本发明涉及此类人工转录因子，其用于影响对脂多糖的细胞应答，以降低或提高Toll样受体4水平，并且用于治疗由脂多糖调节的疾病，尤其用于治疗眼疾病。同样，本发明涉及治疗由脂多糖调节的疾病的方法，其包括向有其需要的患者施用治疗有效量的本发明的人工转录因子。
在另一个具体的实施方案中，本发明涉及人工转录因子中间体，其包含融合至抑制或激活蛋白结构域和核定位序列的特异性靶向Toll样受体4启动子的多指锌指蛋白。
在另一个具体的实施方案中，受体基因启动子是高亲和力免疫球蛋白ε受体亚基α启动子。在另一个具体的实施方案中，本发明涉及此类人工转录因子，其用于影响对免疫球蛋白E（IgE）的细胞应答，以降低或提高高亲和力IgE受体水平，并且用于治疗由IgE调节的疾病，尤其用于治疗眼疾病。同样，本发明涉及治疗由IgE调节的疾病的方法，其包括向有其需要的患者施用治疗有效量的本发明的人工转录因子。
在另一个具体的实施方案中，本发明涉及人工转录因子中间体，其包含融合至抑制或激活蛋白结构域和核定位序列的特异性靶向高亲和力免疫球蛋白ε受体亚基α启动子的多指锌指蛋白。
附图简述
图1：通过调节受体表达改变细胞敏感性
通过蛋白质转导结构域(PTD)诸如TAT或其他的作用将人工转录因子转运至细胞中，所述人工转录因子含有融合至抑制/激活结构域（RD=调节域）以及核定位序列(NLS)的特异性靶向受体基因(RG)启动子(P)的六聚锌指(ZF)蛋白。根据转录调节域，受体基因表达提高(+)或抑制(-)，分别产生增强的或减少的对受体(R1、R2或R3)激动剂(A)的细胞敏感性。
图2：人内皮素受体A（ETRA）启动子区
显示了含有假定的ETRA启动子的ETRA基因的5'非翻译区。突出显示的是转录起始（用+1标记）和人工转录因子潜在的15bp和18 bp靶位点（TS）（下划线表示并标记为TS-855、TS-555、TS-487、TS-447、TS-306、TS-230、TS-103、TS-37、TS+74）。
图3：人Toll样受体4（TLR4）启动子区
显示了含有TLR4启动子的TLR4基因的5'区。突出显示的是转录起始（用+1标记）、第一外显子的起始密码子和可读框（粗体字母）和特异性人工转录因子潜在的18 bp靶位点（下划线表示并标记为TS-276、TS-55、TS+113）。
图4：高亲和力IgE受体A（FCER1A）启动子区
显示了含有邻近的、组成型启动子的FCER1A基因的5'区。突出显示的是转录起始（用+1标记）、第一外显子的起始密码子和可读框（粗体字母）和特异性人工转录因子潜在的18 bp靶位点（下划线表示并标记为TS-147和TS+17）。
图5：人内皮素受体B（ETRB）启动子区
显示了含有ETRB启动子的ETRB基因的5'区。突出显示的是翻译起始（用+1标记）和特异性人工转录因子潜在的18 bp靶位点（下划线表示并标记为TS-1149和TS-487）。由于报道了几个可变转录起始位点(Arai H.等, 1993, J Biol Chem 268, 3463-70; Tsutsumi M.等, 1999, Gene 4, 43-9)，选择翻译起始位点作为命名靶位点的参考点。
图6：人工转录因子
A）将多种锌指蛋白克隆入三个不同质粒中。显示唯一的限制酶位点以突出多种表达质粒的模块设计。产生的DNA构建体编码以下融合蛋白：KRAB-NLS-6ZFP-3xmyc (SEQ ID NO: 8)、SID-NLS-6ZFP-3xmyc、
NLS-6ZFP-GGSGGS (SEQ ID NO: 9) 接头-KRAB A-3xmyc和
NLS-6ZFP-GGSGGS接头-VP64-3xmyc。
图7：通过人工转录因子AO74A、AO74E、AO74R和AO74V调节人内皮素受体A（ETRA）活性
（A）ETRA启动子驱动的蛋白表达的人工转录因子依赖的阻抑。显示了表达指向ETRA启动子内的靶位点的AO74A (SEQ ID NO: 10)、AO74E (SEQ ID NO: 11)、AO74R (SEQ ID NO: 12)和AO74V (SEQ ID NO: 13)后的萤光素酶报道基因测定(RLuA = 相对萤光素酶活性, 相对于对照C，以％计)的结果。C = 作为对照的黄色荧光蛋白(YFP)。
(B) AO74Vp (SEQ ID NO:14), 转导的AO74V蛋白，相比于对照B（缓冲液处理的细胞）不抑制HeLa细胞增殖。RP = 以对照的％计的相对增殖。
(C) AO74Vp相比于对照B（缓冲液处理的细胞）不抑制人子宫平滑肌细胞(hUtSMC)的增殖。
(D) AO74Vp阻断了hUtSMC的ET-1依赖性收缩。将hUtSMC植入三维胶原网格。C = 作为对照的用缓冲液处理的细胞。B = 用缓冲液和ET-1处理的细胞。AO74Vp = 用AO74Vp和ET-1处理的细胞。RLA = 以对照C的％计的相对网格面积。细节描述于下文。
图8：通过人工转录因子AO74Ra和AO74Va驱动的ETRA启动子活性的增强
(A) 激活人工转录因子AO74Ra (SEQ ID NO: 15)和AO74Va (SEQ ID NO: 16)表达后，ETRA启动子驱动的萤光素酶报道基因的表达升高。RLuA = 相对萤光素酶活性，以相对于对照C（YFP）的％计。
(B) 用AO74Vap (SEQ ID NO:17)处理并不抑制hUtSMCs细胞增殖。作为蛋白递送的AO74Vp对hUtSMCs细胞没有毒性，并且并不负面影响细胞增殖。B = 缓冲液处理的细胞。RP = 以对照的％计的相对增殖。
图9：通过AO1149N和AO1149P的人内皮素受体B（ETRB）启动子的阻抑
在萤光素酶报道基因测定中，人工转录因子AO1149N (SEQ ID NO: 18)和AO1149P (SEQ ID NO: 19)的表达相对于YFP（对照C）阻抑了ETRB启动子活性。RLuA = 相对萤光素酶活性，以相对于对照C的％计。
图10：通过AO55B和AO55E的人Toll样受体4（TLR4）活性的调节
(A)在萤光素酶报道基因测定中，AO55B (SEQ ID NO: 20)和AO55E (SEQ ID NO: 21)的表达相对于YFP（对照C）阻断了TLR4启动子活性。RLuA = 相对萤光素酶活性，以相对于对照C的％计。
(B)在巨噬细胞样U937细胞中表达AO55B后，TLR4依赖性的、LPS诱导的白细胞介素(IL)-6的分泌变弱（blunted）。
(C)用AO55Bp (SEQ ID NO:22)处理并不抑制HeLa细胞增殖。RP = 以对照的％计的相对增殖。B = 缓冲液处理的细胞。RP = 以对照的％计的相对增殖。
图11：高亲和力IgE受体通过AO147A调节
(A) AO147A (SEQ ID NO: 23)表达后，高亲和力IgE受体α亚基(FCER1A)启动子驱动的萤光素酶报道基因的表达在大鼠嗜碱RBL-2H3细胞中被抑制。RLuA = 相对萤光素酶活性，以相对于对照C（YFP）的％计。
(B) AO147Ap (SEQ ID NO:24)并不抑制HeLa细胞增殖。B = 缓冲液处理的细胞。RP = 以对照的％计的相对增殖。
C).用AO147Ap处理抑制人IgE与人嗜碱KU812F细胞的结合为80％左右。IgEB = 使用人IgE和用FITC标记的小鼠抗人IgE通过流式细胞术测定的IgE对FCER1的结合能力，以相对于作为对照的缓冲液处理的细胞（B）的％计。
发明详述
本发明涉及人工转录因子，其包含融合至抑制或激活蛋白结构域、核定位序列和蛋白质转导结构域的特异性靶向受体基因启动子的多指锌指蛋白，并且涉及包含此类人工转录因子的药物组合物。
许多疾病的治疗基于调节细胞受体信号传递。实例是高血压（其中β阻断剂抑制β肾上腺素能受体的功能）、抑郁（其中5-羟色胺摄取阻断剂增加激动剂浓度和因此的5-羟色胺受体信号传递）、或青光眼（其中前列腺素类似物活化前列腺素受体，继而降低眼内压）。传统上，为了治疗目的，使用以受体激动剂或拮抗剂形式的小分子来影响受体信号传递。然而，细胞受体信号传递还能够被受体蛋白表达的直接调节所影响。
顺从于受体表达水平的直接调节的病理学过程例如为以下：患有由于先天性心脏病的充血性心力衰竭的患者将受益于β肾上腺素受体的上调，这是由于该受体在心肌中的下调与手术后心力衰竭风险相关。在帕金森氏病中，用多巴胺能药疗法的治疗抑制多巴胺受体的可利用性，因此，多巴胺受体的上调将改善多巴胺能药疗法的效力。在癫痫中，海马中大麻素受体表达不足涉及疾病病因学，因此，大麻素受体的上调将是癫痫患者的可行疗法。
对于由受体蛋白的单倍剂量不足引起的遗传疾病，诸如引起生长迟缓的胰岛素样生长因子I受体，但还有其他的，额外激活剩余的有功能的受体基因将有益于患者。此外并在其他中，病理性自身免疫的诱导和永续性（perpetuation）与从Toll样受体的不恰当信号传递相关联。因此，下调Toll样受体破坏多种自身免疫疾病的恶性循环。在变应性疾病中，通过高亲和力IgE受体防止IgE介导的信号传递用于操纵变应性反应。在癌症中，下调生长因子受体或上调胞外基质受体对于防止肿瘤进展是有利的。
在此类受体分子中的是来自所谓的七跨膜或G蛋白偶联受体（GPCR）蛋白家族的蛋白，其特征在于将受体锚定在质膜中的七跨膜结构域和G蛋白依赖性信号级联放大。此类蛋白的实例是内皮素的受体A和B。其他受体蛋白经单跨膜区锚定，例如脂多糖的受体、Toll样受体4或多种细胞因子受体诸如IL-4受体。其他受体由多聚蛋白复合体组成，例如由α、β和γ链组成的IgE抗体的高亲和力受体，或者由α、β、γ、δ、ε和ζ链组成的T细胞受体。因此，在术语“受体分子”下包含的是来自不同蛋白家族具有非常不同的作用模式的蛋白。
本发明中考虑的受体是人受体分子，其由以下编码：
和KIT。
考虑的进一步的受体是人受体，其识别白细胞介素
，瘦蛋白，干扰素-α，干扰素-β，干扰素-γ，肿瘤坏死因子α，淋巴毒素，促乳素，制瘤素M，白血病抑制因子，集落刺激因子，免疫球蛋白A，免疫球蛋白D，免疫球蛋白G，免疫球蛋白M，免疫球蛋白E，人白细胞抗原（HLA）A，HLA-B，HLA-C，HLA-E，HLA-F，HLA-G，HLA-DP，HLA-DQ，HLA-DR，转化生长因子α，转化生长因子β，神经生长因子，脑源性神经营养因子，神经营养因子-3，神经营养因子-4，肾上腺髓质素，血管生成素，自分泌运动因子，骨形态发生蛋白，促红细胞生成素，成纤维细胞生长因子，神经胶质细胞源性神经营养因子，粒细胞集落刺激因子，粒细胞巨噬细胞集落刺激因子，生长分化因子-9，肝细胞生长因子，肝癌衍生生长因子，胰岛素样生长因子，胰岛素，迁移刺激因子，肌肉生长抑制素（myostatin），血小板衍生生长因子，血小板生成素，血管内皮生长因子，胎盘生长因子和生长激素。
进一步考虑的是由同源非人基因编码的受体，例如由猪、马、牛、猫、犬或鼠基因编码的受体；和由同源植物受体基因编码的受体，例如在作物植物诸如小麦、大麦、玉米、稻、黑麦、燕麦、大豆、花生、向日葵、红花、亚麻、豆类、烟草、或生活饲料草中发现的基因，和在水果植物诸如苹果、梨、香蕉、柑橘类水果、葡萄等中发现的基因。
逆转录病毒具有特别高的免疫原性潜能，因而限制了它们在某一治疗的重复应用中的使用。由于锌指模块的高保守性，这样的免疫反应在应用本发明的人工转录因子后将较小或不存在，或者可能通过消除免疫原性同时仍保留靶位点结合及因此的功能的对整体结构的小改变来避免或进一步降到最低。此外，考虑用聚乙二醇修饰本发明的人工转录因子以降低免疫原性。此外，应用本发明的人工转录因子到免疫赦免器官诸如眼或脑，将避免任何免疫反应，并降低全身对人工转录因子的耐受。对于治疗免疫赦免器官外的慢性疾病，考虑经在前眼球内注射诱导免疫耐受。
调节受体活性的小分子的鉴定主要依赖于在来自不同种类物质的多种多样的不同分子中广泛且耗时的筛选程序。尤其是，对于给定受体分子的此类小分子药物的快速、合理设计是非常有挑战的。相反，本发明的人工转录因子均属于同一物质种类，具有高度确定的总体组成。靶向两条非常不同的启动子序列的两个基于六聚锌指蛋白的人工转录因子仍具有85％的最小氨基酸序列同一性和总体相似的三级结构，并且能够经标准化方法（如下文所述）以快速且经济的方式来生成。因此，本发明的人工转录因子在一类分子中组合了对非常广泛且不同的组的靶格外高的特异性与总体相似的组成。此外，将本发明的人工转录因子配制到药物中可以依赖于进一步加速药物开发进程的先前经验。
人工转录因子的蛋白质转导结构域（PTD）介导的胞内递送是以新方式利用生物制剂对靶受体分子的高选择性的新方法。尽管常规药物调节某些受体的活性，但人工转录因子改变这些蛋白的可利用性。并且由于人工转录因子被设计为特异性作用于此类受体基因的启动子区，因此本发明允许选择性地靶向甚至密切相关的蛋白。这基于甚至密切相关的蛋白的启动子区的仅松散的保守性。蛋白质转导结构域介导的人工转录因子的递送用于调节细胞对外界刺激的反应，所述外界刺激包括但不限于激素如例如胰岛素、内皮素或免疫调节肽诸如白细胞介素、趋化因子和细胞因子，但还有抗体、抗原和分子模式。而且对其他可溶性信号分子诸如谷氨酸或γ-氨基丁酸和其他神经递质的细胞应答可以通过该方法调节。利用本发明的人工转录因子的高选择性，基于给定受体蛋白家族的某些成员的时常组织特异性表达，甚至药物作用的组织特异性靶向也是可能的。
本发明还涉及此类人工转录因子在治疗由特异性效应物与受体的结合所调节的疾病中的用途，为此多指锌指蛋白特异性靶向受体基因启动子。同样，本发明涉及治疗疾病的方法，其包括向有其需要的患者施用治疗有效量的人工转录因子，其中待治疗疾病由特异性效应物对受体的结合所调节，为此多指锌指蛋白特异性靶向受体基因启动子。
所考虑的多指锌指蛋白是四聚的、五聚的、六聚的或七聚的锌指蛋白。“四聚的”、“五聚的”、“六聚的”和“七聚的”表示锌指蛋白分别由四个、五个、六个和七个部分蛋白结构组成，其每一个具有对特定核苷酸三联体的结合特异性。优选地，人工转录因子包含六聚锌指蛋白。
在给定启动子区内选择靶位点
靶位点选择对于成功产生有功能的人工转录因子是至关重要的。对于人工转录因子体内调节靶基因表达，它必须在靶基因的基因组背景中结合它的靶位点。这需要DNA靶位点的可接近性，表示该区域中的染色体DNA并未被围绕组蛋白紧密包装成核小体，并且没有DNA修饰诸如甲基化干扰人工转录因子结合。尽管大部分的人基因组被紧密包装并且是转录无活性的，但活跃转录基因的转录起始位点的紧挨的附近(-1000至+200bp)必须对于内源转录因子和转录机诸如RNA聚合酶是可接近的。因此，在任何给定靶基因的该区域中选择靶位点将极大增强产生具有体内期望功能的人工转录因子的成功率。
在人内皮素受体A（ETRA）启动子区内选择靶位点
通过如下分析人ETRA基因确定特异性靶向内皮素受体A启动子的六聚锌指蛋白（6ZFP）：
人ETRA基因(含启动子区SEQ ID NO: 25、编码区SEQ ID NO: 26的基因组区)由被七个内含子分隔的八个外显子构成(Hosoda K.等, 1992, J Biol Chem 267, 18797-18804)。外显子1和内含子1位于5'非编码区，转录起始位点在ATG翻译起始密码子上游502 bp。
分析相对于转录起始位点从-1000 bp至+100 bp的ETRA启动子区的(GNN)6 靶位点（图2和表1）。使用ZiFiT软件(Sander J.D.等, 2010, Nucleic Acids Res 38, W462-468), 鉴定到TS-855和TS+74，并且选择Barbas组的GNN锌指模块来设计ZFP-855A和ZFP+74A。
尽管基于预先选择的锌指模块的锌指蛋白的合理设计是已知的，但基于含有无偏向ZFP的文库的筛选方法证明对于鉴定高亲和力锌指蛋白是优秀的。因此，选择最初被ZiFiT程序排除的另外的(GNN)6序列(TS-103)。此外，在TS-855和TS+74之间选择含有GNN或CNN三联体的其他18 bp靶位点。此外，在TS-855和TS-306之间选择15 bp靶位点用于筛选6ZFP文库。
在人内皮素受体B（ETRB）启动子区内选择靶位点
如下选择用于调节ETRB表达的人工转录因子的结合位点：ETRB基因的5'区(SEQ ID NO: 27)在翻译起始位点上游的-1195、-817、-229和-258 bp处含有假定的转录起始位点。因此，在-1149 bp和-487 bp之间选择由GNN或CNN三联体组成的18bp靶位点（见图5）。
在人Toll样受体4（TLR4）启动子区内选择靶位点
在TLR4基因的5'区(SEQ ID NO: 28)中在相对于转录起始位点的-276 bp和+113 bp之间选择由六个G/CNN三联体组成的调节TLR4表达人工转录因子的18 bp潜在结合位点（见图3）。
在人高亲和力IgE受体A（FCER1A）启动子区内选择靶位点
在FCER1A基因的5'区(SEQ ID NO: 29)中选择FCER1A表达调节人工转录因子的结合位点。人FCER1A启动子含有在转录起始位点上游200 bp左右的近侧调节区以及进一步上游的含有IL-4应答元件的远侧区(Nishiyama C., 2006, Biosci Biotechnol Biochem 70 (1), 1-9)。在远侧调节区中相对于转录起始位点的-147 bp和+17 bp处选择调节FCER1A人工转录因子的潜在结合位点。
用于选择六聚锌指蛋白的改良的酵母单杂交(Y1H)筛选
基于由Gonzalez B.等, 2010, Nat Protoc 5, 791-810发表的文库克隆方案，将酵母穿梭载体pGAD10 (pAN1025)改良以允许锌指蛋白编码文库有效产生。为了改进克隆效力，在pBluescript中完成锌指蛋白编码文库的最初装配，随后将文库转移至pAN1025中。使用连续消化和DNA去磷酸化，防止形成头-头或尾-尾连接的锌指模块，因而改进了有效的文库覆盖度。
常规Y1H筛选旨在从天然存在蛋白相对小的库中鉴定出给定DNA序列的转录因子。本文的目的在于从均具有结合所用靶位点的潜力的蛋白的非常大的库（16*106个左右）中选择六聚锌指蛋白（6ZFP）。这需要使用额外的选择压力以鉴定具有最高靶位点亲和力的6ZFP。尽管一般将200 ng/ml的金担子素A (AbA)浓度用于常规Y1H分析，但将高至4000 ng/ml AbA用于改进上文用所利用的Y1H系统通常实现的选择(MatchMaker Gold, Clontech)。
为了进一步提高选择压力并因而鉴定对给定靶位点具有高得多的结合能力的6ZFP，进一步改良Y1H系统。对于第一轮选择，将人工转录因子文库包含在基于2 μ复制起点的酵母载体中。此类载体在酵母细胞中独立复制至约50个拷贝，导致6ZFP的强力产生。对于第二轮选择，将人工转录因子文库包含在基于具有1-2/细胞的拷贝数的低拷贝ARS/CEN载体的酵母载体中。由于文库锌指蛋白的较低表达水平，组合有4000 ng/ml的AbA的基于ARS/CEN的Y1H筛选更为灵敏，并且产生对其同族靶序列具有更高结合亲和力的6ZFP。
表1：ETRA启动子区内的靶位点和Y1H筛选的结果
a) 第1栏中显示ETRA启动子靶位点（根据它们与转录起始位点的距离来命名）。
b) 第2栏对在Y1H筛选中鉴定到结合ETRA启动子靶位点的ZFP命名。命名方案如下：ZFP，随后为靶位点的名称和指明在筛选中分离到的不同ZFP的字母。
c) 第3栏根据它们的确定的结合偏好通过详述各个锌指模块显示了ZFP的构成。GM01指明优选结合至GAA三联体的锌指模块, GM02指明优选结合至GCA, GM03指明优选结合至GGA, GM04指明优选结合至GTA, GM05指明优选结合至GAC, GM06指明优选结合至GCC, GM07指明优选结合至GGC, GM08指明优选结合至GTC, GM09指明优选结合至GAG, GM10指明优选结合至GCG, GM11指明优选结合至GGG, GM12指明优选结合至GTG, GM13指明优选结合至GAT, GM14指明优选结合至GCT, GM15指明优选结合至GGT, GM16指明优选结合至GTT, 并且此外, CM01指明优选结合至CAC, CM02指明优选结合至CAA, CM03指明优选结合至CAG, CM04指明优选结合至CAT, CM05指明优选结合至CCA, CM06指明优选结合至CCC, CM07指明优选结合至CCG CM08指明优选结合至CCT, CM09指明优选结合至CGA, CM10指明优选结合至CGC, CM11指明优选结合至CGG, CM12指明优选结合至CGT, CM13指明优选结合至CTA, CM14指明优选结合至CTG, 和CM15指明优选结合至CTT。
d) 第4栏指鉴定到的结合各个靶位点序列的ZFP的序列ID。
表2：ETRB启动子内的靶位点和Y1H筛选的结果
a) 第1栏中显示ETRB启动子靶位点（根据它们与翻译起始位点的距离来命名）。
b) 第2栏对在Y1H筛选中鉴定到结合ETRB启动子靶位点的ZFP命名。命名方案如下：ZEP，随后为靶位点的名称和指明在筛选中分离到的不同ZFP的字母。
c) 第3栏根据它们的确定的结合偏好通过详述各个锌指模块显示了ZFP的构成。GM01指明优选结合至GAA三联体的锌指模块, GM02指明优选结合至GCA, GM03指明优选结合至GGA, GM04指明优选结合至GTA, GM05指明优选结合至GAC, GM06指明优选结合至GCC, GM07指明优选结合至GGC, GM08指明优选结合至GTC, GM09指明优选结合至GAG, GM10指明优选结合至GCG, GM11指明优选结合至GGG, GM12指明优选结合至GTG, GM13指明优选结合至GAT, GM14指明优选结合至GCT, GM15指明优选结合至GGT, GM16指明优选结合至GTT, 并且此外, CM01指明优选结合至CAC, CM02指明优选结合至CAA, CM03指明优选结合至CAG, CM04指明优选结合至CAT, CM05指明优选结合至CCA, CM06指明优选结合至CCC, CM07指明优选结合至CCG CM08指明优选结合至CCT, CM09指明优选结合至CGA, CM10指明优选结合至CGC, CM11指明优选结合至CGG, CM12指明优选结合至CGT, CM13指明优选结合至CTA, CM14指明优选结合至CTG, 和CM15指明优选结合至CTT。
d) 第4栏指鉴定到的结合各个靶位点序列的ZFP的序列ID。
表3：TLR4启动子内的靶位点和Y1H筛选的结果
a) 第1栏中显示TLR4启动子靶位点（根据它们与转录起始位点的距离来命名）。
b) 第2栏对在Y1H筛选中鉴定到结合TLR4启动子靶位点的ZFP命名。命名方案如下：ZFP，随后为靶位点的名称和指明在筛选中分离到的不同ZFP的字母。
c) 第3栏根据它们的确定的结合偏好通过详述各个锌指模块显示了ZFP的构成。GM01指明优选结合至GAA三联体的锌指模块, GM02指明优选结合至GCA, GM03指明优选结合至GGA, GM04指明优选结合至GTA, GM05指明优选结合至GAC, GM06指明优选结合至GCC, GM07指明优选结合至GGC, GM08指明优选结合至GTC, GM09指明优选结合至GAG, GM10指明优选结合至GCG, GM11指明优选结合至GGG, GM12指明优选结合至GTG, GM13指明优选结合至GAT, GM14指明优选结合至GCT, GM15指明优选结合至GGT, GM16指明优选结合至GTT, 并且此外, CM01指明优选结合至CAC, CM02指明优选结合至CAA, CM03指明优选结合至CAG, CM04指明优选结合至CAT, CM05指明优选结合至CCA, CM06指明优选结合至CCC, CM07指明优选结合至CCG CM08指明优选结合至CCT, CM09指明优选结合至CGA, CM10指明优选结合至CGC, CM11指明优选结合至CGG, CM12指明优选结合至CGT, CM13指明优选结合至CTA, CM14指明优选结合至CTG, 和CM15指明优选结合至CTT。
d) 第4栏指鉴定到的结合各个靶位点序列的ZFP的序列ID。
表4：FCER1A启动子内的靶位点和Y1H筛选的结果
a) 第1栏中显示FCER1A启动子靶位点（根据它们与转录起始位点的距离来命名）。
b) 第2栏对在Y1H筛选中鉴定到结合FCER1A启动子靶位点的ZFP命名。命名方案如下：ZFP，随后为靶位点的名称和指明在筛选中分离到的不同ZFP的字母。
c) 第3栏根据它们的确定的结合偏好通过详述各个锌指模块显示了ZFP的构成。GM01指明优选结合至GAA三联体的锌指模块, GM02指明优选结合至GCA, GM03指明优选结合至GGA, GM04指明优选结合至GTA, GM05指明优选结合至GAC, GM06指明优选结合至GCC, GM07指明优选结合至GGC, GM08指明优选结合至GTC, GM09指明优选结合至GAG, GM10指明优选结合至GCG, GM11指明优选结合至GGG, GM12指明优选结合至GTG, GM13指明优选结合至GAT, GM14指明优选结合至GCT, GM15指明优选结合至GGT, GM16指明优选结合至GTT, 并且此外, CM01指明优选结合至CAC, CM02指明优选结合至CAA, CM03指明优选结合至CAG, CM04指明优选结合至CAT, CM05指明优选结合至CCA, CM06指明优选结合至CCC, CM07指明优选结合至CCG CM08指明优选结合至CCT, CM09指明优选结合至CGA, CM10指明优选结合至CGC, CM11指明优选结合至CGG, CM12指明优选结合至CGT, CM13指明优选结合至CTA, CM14指明优选结合至CTG, 和CM15指明优选结合至CTT。
d) 第4栏指鉴定到的结合各个靶位点序列的ZFP的序列ID。
本发明的人工转录因子包含基于表1-4第3栏中显示的锌指模块组成的锌指蛋白，其中高至三个单独的锌指模块与具有可选结合特征的其他锌指模块交换，以调节人工转录因子与它的靶序列的结合。
本发明的人工转录因子包含基于表1-4第3栏中显示的锌指模块组成的锌指蛋白，其中交换单独的氨基酸以将潜在的免疫原性降到最低，同时保留对预期靶位点的结合亲和力。
优选地，本发明的人工转录因子包含具有选自以下的蛋白序列的锌指蛋白：SEQ ID NO:31至SEQ ID NO:37, SEQ ID NO:39至SEQ ID NO:43, SEQ ID NO:45至SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54至SEQ ID NO:57, SEQ ID NO:59至SEQ ID NO:64, SEQ ID NO:66至SEQ ID NO:80, SEQ ID NO:82至SEQ ID NO:95, SEQ ID NO:97至SEQ ID NO:118, SEQ ID NO:120至SEQ ID NO:136, SEQ ID NO:138至SEQ ID NO:143, SEQ ID NO:145至SEQ ID NO:153, SEQ ID NO:155至SEQ ID NO:164, SEQ ID NO:166至SEQ ID NO:173, SEQ ID NO:175至SEQ ID NO:181, 和SEQ ID NO:183至SEQ ID NO:191。
更优选地，本发明的人工转录因子包含具有SEQ ID NO 135的五聚锌指蛋白或具有选自以下蛋白序列的六聚锌指蛋白：SEQ ID NO 33, 54, 56, 64, 68, 83, 84, 85, 97, 101, 114, 118, 122, 127, 133, 140, 142, 146, 147, 156, 159, 169, 171, 173, 175, 181, 184, 187, 189, 和191。
甚至更优选的是人工转录因子，其包含具有SEQ ID NO 135的五聚锌指蛋白或具有选自以下蛋白序列的六聚锌指蛋白：SEQ ID NO 56, 83, 85, 101, 114, 118, 127, 133, 140, 142, 146, 147, 156, 159, 175, 和181。
甚至更优选的是人工转录因子，其包含具有SEQ ID NO 118, 127, 146, 156, 或175的六聚锌指蛋白。
甚至更优选的是人工转录因子，其包含具有SEQ ID NO 118, 127, 156, 或175的六聚锌指蛋白。
最优选的是人工转录因子，其包含具有SEQ ID NO 118, 156, 或175的六聚锌指蛋白。
将多指锌指蛋白融合至调节域，其是抑制或激活蛋白结构域。考虑的抑制蛋白结构域是由基因本体论GO:0001071确定的蛋白的转录活性结构域，诸如 N-末端KRAB、C-末端KRAB、SID和ERD结构域、优选KRAB或SID。考虑的激活蛋白结构域是由基因本体论GO:0001071确定的蛋白的转录活性结构域，诸如VP16或VP64 (VP16的四聚体重复)，优选VP64。
进一步对于融合至抑制或激活蛋白结构域的多指锌指蛋白，本发明的人工转录因子包含核定位序列（NLS）。考虑的核定位序列是通过结合至由基因本体论GO:0008139确定的蛋白而赋予核输入的氨基酸基序，例如碱性氨基酸的簇，其包含赖氨酸残基，随后为赖氨酸或精氨酸残基，随后为任何氨基酸，随后为赖氨酸或精氨酸残基(K-K/R-X-K/R共有序列, Chelsky D.等, 1989 Mol Cell Biol 9, 2487-2492)或SV40 NLS，并且SV40 NLS是优选的。
本发明的人工转录因子进一步任选包含蛋白质转导结构域（PTD）。考虑的蛋白质转导结构域是HIV衍生的TAT肽、HSV-1 VP22肽、合成肽mT02 (PVRRPRRRRRRK, SEQ ID NO: 192, Yoshikawa T.等 2009 Biomaterials 30, 3318-23)、合成肽mT03 (THRLPRRRRRRK, SEQ ID NO: 193)、R9肽(RRRRRRRRR, SEQ ID NO: 194)、ANTP结构域和保护性抗原/致死因子N末端PTD，优选TAT PTD。
指向受体基因启动子但不具有蛋白质转导结构域的人工转录因子也是本发明的主题。它们是如上文确定的本发明的人工转录因子的中间体。
指向ETRA但不具有蛋白质转导结构域的人工转录因子也是本发明的主题。它们是如上文确定的本发明的人工转录因子的中间体。
指向ETRB启动子但不具有蛋白质转导结构域的人工转录因子也是本发明的主题。它们是如上文确定的本发明的人工转录因子的中间体。
指向TLR4启动子但不具有蛋白质转导结构域的人工转录因子也是本发明的主题。它们是如上文确定的本发明的人工转录因子的中间体。
指向FCER1A启动子但不具有蛋白质转导结构域的人工转录因子也是本发明的主题。它们是如上文确定的本发明的人工转录因子的中间体。
本发明的人工转录因子的结构域可以通过短的柔性接头连接。短的柔性接头具有2-8个氨基酸，优选甘氨酸和丝氨酸。考虑的具体接头是GGSGGS (SEQ ID NO:9)。人工转录因子还可以包含标志物以便于它们的检测和加工。
人工转录因子在调节受体启动子活性中的活性
根据图6中显示的方案从使用Y1H筛选选择的特异性结合受体启动子的某些靶位点的ZFP（见表1-4）构建基于锌指模块的人工转录因子。这些人工转录因子包含不同的转录活性结构域诸如N-末端KRAB、C-末端KRAB、SID或VP64。根据发表的数据(Beerli R.R.等, 1998 Proc Natl Acad Sci USA 95, 14628-14633)，预测KRAB以及SID结构域作为转录阻抑蛋白起作用，而VP64介导转录激活。为了评估人工转录因子（6ZFP和转录活性结构域之间的融合体）影响由受体启动子驱动的转录的潜力，使用萤光素酶报道基因测定。为此目的，用人工转录因子表达质粒与双报道基因质粒一同共转染能够从某一启动子驱动表达的细胞。双报道基因质粒包含在所讨论的受体启动子的控制下的分泌型Gaussia萤光素酶基因以及在基于NEG-PG04和EF1a-PG04质粒(GeneCopoeia, Rockville, MD)的组成型CMV启动子的控制下的分泌型碱性磷酸酶(SEAP)的基因。
许多启动子展示出细胞类型特异性表达模式，在一些细胞类型中实际上无表达，并且在其他细胞类型中高水平表达。因此，对于启动子调节研究选择合适细胞模型依赖于给定受体启动子的组织特异性。在本文显示的例子中，子宫颈癌细胞系HeLa细胞能够从ETRA、ETRB或TLR4启动子表达萤光素酶报道基因。由于FCER1A启动子的组织特异性，因此在该启动子的控制下萤光素酶的表达在HeLa细胞中是不可能的。因此，利用大鼠嗜碱细胞性白血病RBL-2H3细胞来评估针对FCER1A启动子的人工转录因子功能。该细胞系支持FCER1A启动子驱动的萤光素酶报道基因的表达，并可以使用核转染以50％左右的效力转染。
以3:1的ATF:报道基因质粒的比例完成共转染，以确保在用报道基因质粒和萤光素酶转染的细胞中人工转录因子（ATF）表达的存在，并且根据制造商建议(GeneCopoeia, Rockville, MD)测量SEAP活性。将萤光素酶值对SEAP活性进行归一化，并与设定为100％的表达黄色荧光蛋白（YFP）的对照细胞进行比较。通过测量经转染的细胞的上清液中萤光素酶与SEAP活性之间的比例，仅在转染有人工转录因子质粒的细胞中受体启动子驱动的萤光素酶表达对SEAP表达的归一化是可能的。该方法证明对于计算并归一化不同实验之间的转染效力中的差异是有用的，并且允许定量人工转录因子介导的给定受体启动子的调节。
将所有萤光素酶表达研究（图7A-11A）进行至少三次，一式三份，进行平均化，与对照转染细胞比较，表示为以对照的％计的相对萤光素酶活性(RLuA)，并作图，其具有表示SEM的误差棒。
人工转录因子的命名约定如下：将这样的人工转录因子用字母AO随后为代表靶位点的数字和辨识使用Y1H筛选鉴定的某一ZFP的字母来指定，所述人工转录因子使用哺乳动物表达载体表达于哺乳动物细胞中，并由锌指蛋白（ZFP）、核定位序列和负调节结构域诸如SID或KRAB（N-或C-末端）组成。向该名称添加的小写的“a”指代含有激活VP64结构域的人工转录因子。添加的小写“p”指代纯化的人工转录因子蛋白，其产生于异源表达系统中，并且除了上述结构域之外，还含有蛋白质转导结构域TAT和HA标签(SEQ ID NO: 195)。
图7A显示了ETRA启动子依赖性萤光素酶表达的人工转录因子依赖性下调。用如上所述的ETRA启动子萤光素酶/组成型SEAP报道基因构建体和AO74A、AO74E、AO74R、AO74V或作为对照（标记为C）的黄色荧光蛋白（YFP）的表达质粒共转染HeLa细胞。这些人工转录因子指向ETRA启动子的TS+74，并且含有负调节的SID结构域。尽管AO74A和AO74E抑制ETRA启动子驱动的表达约70％，但AO74R和AO74V能够阻断ETRA启动子至背景水平。
图8A突出显示了用于生成靶向受体启动子的转录因子的方法的多样性。通过简单交换AO74V或AO74R中的抑制结构域SID为激活结构域VP64，可以产生能够增强ETRA启动子的转录活性至400％左右的激活转录因子。
使用相同方法，构建靶向ETRB受体启动子的人工转录因子。图9A显示了由含有指向ETRB启动子的靶位点TS-1149的ZFP（见图2）以及抑制SID结构域的AO1149N和AO1149P的ETRB启动子活性的抑制。用ETRB启动子萤光素酶/SEAP报道基因构建体和AO1149N、AO1149P或作为对照的YFP的表达质粒共转染HeLa细胞。AO1149N抑制ETRB启动子活性80％左右，而AO1149P阻断ETRB启动子几乎至背景水平。
为了分析由指向TLR4启动子中的靶位点TS-55的ZFP（见图3）和在C-末端的抑制KRAB结构域组成的TLR4特异性人工转录因子AO55B和AO55E的活性，使用Gaussia萤光素酶/SEAP报道基因测定。如图10中显示，在HeLa细胞中AO55B或AO55E的表达抑制TLR4启动子驱动的表达，并且相比于表达YFP的对照转染的细胞，AO55B完全阻断了萤光素酶表达。
为了评估指向FCER1A启动子的人工转录因子的活性，将AO147A与FCER1A启动子驱动的Gaussia萤光素酶和如上的CMV驱动的SEAP一同表达于大鼠嗜碱RBL-2H3细胞中。该人工转录因子指向靶位点TS-147，并且含有N-末端KRAB结构域。基于FCER1A启动子的组织特异性和使用核穿孔（nucleoporation）转染的方便性，选择RBL-2H3细胞。如图11所示，在产生AO147A的RBL-2H3细胞中FCER1A驱动的表达相比于表达YFP的对照细胞（C）降低80％左右。
总之，依赖于用于人工转录因子构建的调节结构域，受体启动子驱动的调节的人工转录因子介导的调节是可行的，并且能够上调表达高至400％，或者完全阻断表达，这开发了以几乎两个数量级的级别的可能调节范围。
评估人工转录因子的潜在毒性作用
尽管对于给定的靶序列选择人工转录因子，并且尽管所选的靶位点在人基因组内是唯一的，但人工转录因子可能通过结合类似序列而具有脱靶作用，由此产生毒性作用。此类毒性作用可能潜在地干扰此类人工转录因子的功能测定。对于任何给定的唯一的18 bp靶位点，可以鉴定到具有一个、两个或三个取代的任何数目的高度相似序列。尽管这些序列可能允许人工转录因子的结合并且可能导致脱靶作用，但大部分此类脱靶位点位于其他位置而非活跃转录基因的调控序列内，极大地改善了人工转录因子处理的脱靶作用的潜力。对于以下实验，用1 μM转导的人工转录因子蛋白处理细胞，并使用MTS测定评估细胞增殖作为测量潜在毒性。每一实验进行一式三份至少三次；将人工转录因子处理后的细胞的增殖进行平均化，并表示为用相应缓冲液处理的对照的百分比。值得注意的是，向人工转录因子指代中加入“p”表示含有TAT蛋白质转导结构域、锌指蛋白和调节结构域诸如SID、KRAB或VP64的蛋白，而非编码不具有蛋白质转导结构域的人工转录因子的表达质粒。
如图7B中显示，用1 μM的AO74Vp蛋白（AO74V的转导版本）处理HeLa细胞两天，相对于用缓冲液处理的细胞并未引起增殖损失。类似地，用AO74Vp处理hUtSMC对于这些细胞没有毒性，并且不抑制它们的增殖（图7C）。此外，基于ZFP-74V的激活人工转录因子蛋白AO74Vap不展示出对hUtSMC增殖的任何毒性作用（图8B）。这些数据与含有ZFP-74V的人工转录因子在人基因组中的可忽略的脱靶结合是一致的。类似地，用TLR-4特异性人工转录因子AO55Bp（图10C）或FCER1A特异性的AO147Ap（图11B）处理HeLa细胞不会导致增殖损失。总之，所有检测的人工转录因子对于它们预期靶位点是高度特异性的，并且不会引起可能导致细胞死亡或增殖降低的脱靶作用。
使用细胞应答测定的人工转录因子功能分析
为了确定基于TAT的蛋白递送后人工转录因子对内源受体启动子的功能，观察人工转录因子和对照处理的细胞之间对受体激动剂处理的细胞应答。
人工转录因子处理后ETRA下调的评估
平滑肌细胞（SMC）表达ETRA并且能够在暴露于ET-1后收缩。为了测量抗ETRA启动子人工转录因子AO74V的有效性，使用人子宫平滑肌细胞(hUtSMC)作为模型系统。为此目的，将hUtSMC植入三维胶原网格并在暴露于0或100 nM ET-1之前用1 μM AO74Vp或缓冲液对照处理三天。每24小时重复蛋白或缓冲液处理。在网格从它们的支持物上脱离下并加入ET-1后，观察到网格的收缩。如图7D中所示，相比于未用ET-1处理的网格，暴露于ET1的对照网格收缩约78％。相反，当与未用ET-1处理的对照网格相比时，AO74V处理的网格在ET-1存在的情况下没有明显收缩。这与用AO74Vp处理后完全阻断ET-1诱导的hUtSMC的收缩是一致的。图7D中显示的数据代表一式六份完成的三次独立实验的加入ET-1后9小时的平均网格面积。使用SPSS软件包利用一般线性单变量模型的统计学分析揭示了AO74Vp的阻断作用的高显著性(** 代表p<0.001)。
人工转录因子处理后TLR4下调的评估
巨噬细胞表达TLR4并且响应LPS结合至TLR4来产生促炎性细胞因子诸如IL-6。佛波醇-12-肉豆蔻酸酯-13-乙酸酯(PMA)刺激的U937细胞是人巨噬细胞样细胞的广泛接受的模型。为了测量抗TLR4启动子人工转录因子AO55B的有效性，用0.5 ng/ml LPS攻击表达AO55B或作为对照的YFP的PMA刺激的U937细胞8小时，并且使用ELISA测量IL-6的产生。如图10B中所示，相比于对照细胞，AO55B的表达显著降低(p< 0.005) IL-6的分泌25％左右。考虑到U937核转染效力约为50％，表示AO55B在这些实验中仅在约50％的细胞中表达，由AO55B对IL-6产生的实际抑制在50％的级别内。
人工转录因子处理后FCER1功能的评估
IgE抗体结合巨噬细胞表面上的异源三聚的高亲和力IgE受体FCER1时，肥大细胞和嗜碱细胞是在特应性个体中引发变应性应答的第一步。遇到变应原导致IgE装载的FECR1分子的交联，引发胞内信号级联放大，导致变应性介体和细胞因子的释放。因此，IgE结合例如嗜碱细胞的能力是变应性过程中的一个关键步骤。为了评估用指向FCER1的α亚基启动子的人工转录因子处理后IgE的结合能力，用1 μM的AO147Ap或缓冲液日常处理人嗜碱KU812F细胞48小时。处理后，使用流式细胞术测量IgE结合能力。三次独立实验的AO147A处理后的KU812F细胞的平均IgE结合能力（IgEB）显示在图11C中。相比于对照处理后的细胞，用AO147Ap处理降低嗜碱细胞的IgE结合能力约80％。有趣的是，尽管人工转录因子AO147Ap仅指向FCER1受体复合体的α亚基，但整个受体在结合IgE能力方面的功能极大降低。因此，预期变应原诱导的FCER1交联极大降低，提高了变应性介体释放的阈值。与一些其他受体不同，FCER1是由α、β和γ亚基构成的多聚蛋白复合体，所述亚基由三个不同遗传基因座编码。只有含有一个α、一个β和两个γ链（并且α链提供IgE结合位点）的正确装配的FCER1能够引发变应性应答。因此，例如用合适的人工转录因子下调FCER1 α链(FCER1A)的表达将阻止作为整体的FCER1的正确装配和功能。这种观点被其中消除了过敏反应的FCER1A-/-小鼠支持(Dombrowicz D., 1993, Cell 75, 969-976)。因此，使用人工转录因子技术靶向FCER1A表达适合于根除变应性反应。此外，尽管人工转录因子对于一种靶基因是高度特异性的，但多聚受体通常顺从于人工转录因子介导的敲低。
药物组合物
本发明还涉及包含如上定义的人工转录因子的药物组合物。考虑的药物组合物是向温血动物尤其是人肠胃外全身施用的组合物，尤其是静脉内施用的组合物，吸入的组合物，和局部施用的组合物，尤其是眼局部使用，例如作为滴眼剂，或玻璃体内、结膜下、眼球侧（parabulbar）或眼球后施用。尤其优选的是滴眼剂和玻璃体内、结膜下、眼球侧或眼球后施用的组合物。组合物包含单独的活性组分，或者优选与药学上可接受的载体一同的活性组分。进一步考虑的是缓释制剂。活性组分的剂量依赖于待治疗的疾病并依赖于物种、它的年龄、体重和个体病况、个体药代动力学数据以及施用方式。
进一步考虑的是用于口腔递送的药物组合物，尤其是包含合适地装入胶囊的活性组分的或者另外针对消化道降解进行保护的组合物。例如，此类药物组合物可以包含膜透性增强剂、蛋白酶抑制剂，并且由肠溶包衣包封。
药物组合物包含大约1％至大约95％的活性成分。单位剂量形式例如是安瓿、药瓶、吸入器、滴眼剂等。
本发明的药物组合物以本身已知的方式制备，例如通过常规混合、溶解或冷冻干燥方法。
优选给予使用活性组分的溶液、以及悬浮液或分散液，尤其是等渗水溶液、分散液或悬浮液，其例如在含有单独的活性组分或者与载体例如甘露醇一同的活性组分的冷冻干燥的组合物的情况下，可以在使用前配制。药物组合物可以进行灭菌和/或可以包含赋形剂，例如防腐剂、稳定剂、湿润剂和/或乳化剂、增溶剂、用于调节渗透压的盐类和/或缓冲液，并且以本身已知的方式制备，例如通过常规溶解和冷冻干燥方法。所述溶液或悬浮液可以包含增粘剂，具体为羧甲基纤维素钠、羧甲基纤维素、葡聚糖、聚乙烯吡咯酮或明胶、或还有增溶剂，例如Tween 80? (聚氧乙烯(20)脱水山梨醇单油酸酯)。
油中悬浮液包含习惯用于注射目的的植物油、合成油或半合成油作为油组分。关于这一点，可以特别提及液体脂肪酸酯，其包含具有8-22个、尤其12-22个碳原子的长链脂肪酸作为酸组分。这些脂肪酸酯的醇组分具有最多6个碳原子，并且是一价醇或多价醇，例如一价、二价或三价醇，尤其是乙二醇和丙三醇。作为脂肪酸酯的混合物，植物油诸如棉子油、杏仁油、橄榄油、蓖麻油、芝麻油、大豆油和花生油尤其有用。
可注射制剂的制造通常在无菌条件下进行，如例如装入安瓿或药瓶和容器的密封。
对于肠胃外使用，以水可溶性形式的活性组分的水溶液，例如水可溶性盐的水溶液，或者含有增粘物质例如羧甲基纤维素钠、山梨醇和/或葡聚糖（并且如需要的话含有稳定剂）的含水注射悬浮液，是尤其合适的。活性组分任选与赋形剂一同还可以以冷冻干燥剂的形式，并且可以在肠胃外施用前通过加入合适的溶剂配制入溶液中。
用于吸入的组合物可以以气溶胶形式如喷雾剂、雾或以滴剂的形式施用。气溶胶从溶液或悬浮液制备，其可以用剂量测定的吸入器或雾化器（即使用合适的推进剂例如二氯二氟甲烷、三氯氟甲烷、二氯四氟乙烷、二氧化碳或其他合适气体向呼吸道或肺递送特定量的药剂的装置）以由患者吸入的气溶胶化的药物的短爆发（short burst）的形式来递送。还可以提供具有合适粉末基质（base）诸如乳糖或淀粉的用于吸入的粉末喷剂。
滴眼剂优选是含有合适的试剂以使组合物与泪液等渗(295-305 mOsm/l)的活性组分的等渗水溶液。考虑的试剂是氯化钠、柠檬酸、甘油、山梨醇、甘露醇、乙二醇、丙二醇、葡萄糖等。此外，组合物包含缓冲剂，例如磷酸盐缓冲剂、磷酸盐-柠檬酸盐缓冲剂或Tris缓冲剂（三(羟甲基)氨基甲烷），以保持5-8的pH，优选7.0-7.4。组合物还可以含有抗菌性防腐剂，例如对羟基苯甲酸酯类、季铵盐类、诸如苯扎氯铵、聚六亚甲基双胍(PHMB)等。滴眼剂还可以含有黄原胶以产生凝胶样滴眼剂，和/或其他增粘剂，诸如透明质酸、甲基纤维素、聚乙烯醇或聚乙烯吡咯酮。
人工转录因子在治疗方法中的用途
此外，本发明涉及如上所述的指向内皮素受体A启动子的人工转录因子，其用于影响对内皮素的细胞应答，以降低或提高内皮素受体水平，并且用于治疗由内皮素调节的疾病，尤其用于治疗此类眼疾病。同样，本发明涉及治疗由内皮素调节的疾病的方法，其包括向有其需要的患者施用治疗有效量的指向内皮素受体A启动子的人工转录因子。
由内皮素调节的疾病例如是心血管疾病，诸如原发性高血压、肺动脉高压、慢性心脏衰竭以及慢性肾衰竭。此外，通过将内皮素应答变弱来实现在不透射线材料应用之前、过程中和之后的肾保护。此外，多发性硬化受内皮素系统负面影响。
进一步的由内皮素调节的疾病是糖尿病性肾疾病或眼疾病，诸如青光眼性神经变性、眼部血液循环中的血管功能失调、视网膜静脉阻塞、视网膜动脉阻塞、黄斑水肿、年龄相关性黄斑变性、视神经病变、中心性浆液性脉络膜视网膜病变、视网膜色素变性、Susac综合征和Leber遗传性视神经病变。
同样，本发明涉及治疗由内皮素调节的疾病的方法，其包括向有其需要的患者施用治疗有效量的本发明的人工转录因子。尤其是本发明涉及治疗青光眼性神经变性、眼部血液循环中的血管功能失调的方法，尤其是涉及治疗视网膜静脉阻塞、视网膜动脉阻塞、黄斑水肿、视神经病变、中心性浆液性脉络膜视网膜病变、视网膜色素变性和Leber遗传性视神经病变的方法，其包括向有其需要的患者施用有效量的本发明的人工转录因子。本发明的人工转录因子的有效量依赖于待治疗的疾病的具体类型并依赖于物种、它的年龄、体重和个体病况、个体药代动力学数据以及施用方式。对于向眼中施用，优选0.5-1 mg的每月玻璃体注射。对于全身应用，优选10mg/kg的每月注射。此外，还优选将缓释沉积物（slow release deposits）植入眼的玻璃体中。
此外，本发明涉及如上所述的指向内皮素受体B启动子的人工转录因子，其用于影响对内皮素的细胞应答，以降低或提高内皮素受体水平，并且用于治疗由内皮素调节的疾病，尤其用于治疗此类眼疾病。同样，本发明涉及治疗由内皮素调节的疾病的方法，其包括向有其需要的患者施用治疗有效量的指向内皮素受体B启动子的人工转录因子。
由ET-1依赖性的、ETRB介导的人工转录因子调节的疾病是某些癌症、神经变性和炎症相关病症。
此外，本发明涉及如上所述的指向TLR4启动子的人工转录因子，其用于影响对LPS的细胞应答，以降低或提高TLR4水平，并且用于治疗由LPS调节的疾病，尤其用于治疗此类眼疾病。同样，本发明涉及治疗由LPS调节的疾病的方法，其包括向有其需要的患者施用治疗有效量的指向TLR4启动子的人工转录因子。由LPS调节的疾病是类风湿性关节炎、动脉粥样硬化、银屑病、克罗恩病、葡萄膜炎、接触镜相关性角膜炎、角膜炎症、癌症对化学疗法的抗性等。
此外，本发明涉及如上所述的指向FCER1A启动子的人工转录因子，其用于影响对IgE或IgE-抗原复合体的细胞应答，以降低或提高FCER1水平，并且用于治疗由IgE或IgE-抗原复合体调节的疾病，尤其用于治疗此类眼疾病。
同样，本发明涉及治疗由IgE或IgE-抗原复合体调节的疾病的方法，其包括向有其需要的患者施用治疗有效量的指向FCER1A启动子的人工转录因子。由IgE或IgE-抗原复合体调节的疾病是变应性鼻炎、哮喘、湿疹和过敏反应等。
人工转录因子在植物中的用途
此外，本发明涉及靶向植物受体的人工转录因子的用途。优选地，将编码人工转录因子的DNA克隆入用于转化植物定居微生物或植物的载体中。或者，将人工转录因子以用于向植物局部应用的合适组合物直接应用。
实施例
DNA质粒的克隆
对于所有克隆步骤，限制性内切核酸酶和T4 DNA连接酶均购自 New England Biolabs。虾碱性磷酸酶（SAP）来自Promega。将高保真Platinum Pfx DNA聚合酶 (Invitrogen)用于所有标准PCR反应中。DNA片段和质粒按照制造商说明书使用NucleoSpin Extract II试剂盒、NucleoSpin Plasmid试剂盒或NucleoBond Xtra Midi Plus试剂盒(Macherey-Nagel)来分离。寡核苷酸购自Sigma-Aldrich。新生成质粒的所有相关DNA序列通过测序(Microsynth)来验证。
两种六聚锌指蛋白（ZFP-855A和ZFP+74A）的设计和克隆
为了生成调节ETRA表达的人工转录因子，设计由TAT-KRAB-ZFP组成的融合蛋白，并通过基因合成获得对应的、密码子优化的DNA序列。对于融合蛋白的ZFP部分，使用具有对“模块装配”的参数集的ZiFiT软件(Sander J.D.等, 2010, Nucleic Acids Res 38, W462-468; 2007, Nucleic Acids Res 35, W599-605)用所谓的“Barbas模块”集来筛选人ETRA启动子区(相对于转录起始位点的-1000 bp至+100 bp; RefSeq DNA NG_013343)的潜在的(GNN)6 6ZFP靶位点。对于旨在结合靶位点-855（在相对于转录起始位点的-855 bp处开始）的ZFP-855A，按照Wright D.A.等, 2006, Nat Protoc 1, 1637-1652构建ZF59-ZF59-ZF72-ZF58-ZF71-ZF67。类似地，对于旨在结合靶位点+74（相对于转录起始位点的+74 bp）的ZFP+74A装配ZF65-ZF62-ZF58-ZF65-ZF59-ZF59。
作为人工转录因子的阻抑蛋白结构域，选择由人KOX1蛋白的氨基酸1-97组成的KRAB结构域(Beerli, R.R.等, 1998, Proc Natl Acad Sci USA 95, 14628-14633)。对于核靶向，将氨基酸PKKKRKV (SEQ ID NO: 196) (对应于SV40 NLS)以及YKDDDDK (SEQ ID NO: 197) (FLAG标签)整合入融合蛋白中。将编码XhoI-NcoI-KRAB-NLS-FLAG-SpeI-ZFP-855A-HindIII和SpeI-ZFP+74A-HindIII (以用ZFP+74A代替ZFP-855A)的合成基因进行密码子优化并由GenScript合成。在用XhoI/HindIII切割插入片段和载体后，将KRAB-NLS-FLAG- ZFP-855A和KRAB-NLS-FLAG- ZFP+74A插入pcDNA3(-) (Invitrogen)中，分别产生pAN1021和pAN1022。
六聚锌指蛋白文库向猎物质粒的克隆
按照Gonzalez B.等,. 2010, Nat Protoc 5, 791-810并具有以下改进来克隆几个含有结合GNN和/或CNN的锌指(ZF)模块的六聚锌指蛋白文库。合成编码GNN和CNN ZF模块的DNA序列并插入pUC57 (GenScript)中，分别产生pAN1049和pAN1073。在pBluescript SK (+)载体中完成ZFP文库的逐步装配。为了避免在每一单独克隆步骤中插入多个ZF模块导致无功能的蛋白，将pBluescript (及其含有1ZFP、2ZFP或3ZFP的衍生产物)和pAN1049或pAN1073首先与一种限制酶孵育，并随后用SAP处理。在加入第二种限制性内切核酸酶之前，通过NucleoSpin Extract II试剂盒将酶去除。
通过用XhoI、SAP并随后用SpeI处理5 μg pBluescript完成pBluescript-1ZFPL的克隆。通过将10 μg pAN1049 (16种不同GNN ZF模块的释放物)或pAN1073 (15种不同CNN ZF模块的释放物)与SpeI、SAP并随后与XhoI孵育来生成插入片段。为了生成pBluescript-2ZFPL和pBluescript-3ZFPL，将7 μg pBluescript-1ZFPL或pBluescript-2ZFPL用AgeI切割，去磷酸化，并用SpeI切割。通过分别向10 μg pAN1049或pAN1073应用SpeI、SAP和随后XmaI来获得插入片段。通过用AgeI、SAP并随后用SpeI处理6 μg的pBluescript-3ZFPL以获得切割后的载体，来完成pBluescript-6ZFPL的克隆。通过用SpeI、SAP和随后XmaI孵育从pBluescript-3ZFPL释放3ZFPL插入片段。
使用200 ng切割后的载体、400 U T4 DNA连接酶在20 μl总体积中在RT（室温）下过夜并以3:1摩尔比的插入片段:载体设定含有一个、两个和三个ZFP的文库的连接反应。六聚锌指蛋白文库的连接反应包括在200 μl总体积中的2000 ng pBluescript-3ZFPL、500 ng 3ZFPL插入片段、4000 U T4 DNA连接酶，将所述总体积分为10个20 μl，并分别在RT下孵育过夜。根据每一文库所需的克隆数通过几种方法将连接反应的部分转化入大肠杆菌（Escherichia coli）中。为了生成pBluescript-1ZFPL和pBluescript-2ZFPL，将3 μl的连接反应直接用于大肠杆菌NEB 5-α的热激转化。在转化入电感受态大肠杆菌NEB 5-α(来自EquiBio的EasyjecT Plus电穿孔仪, 2.5 kV和25 μF, 来自Bio-Rad的2 mm电穿孔杯)前，通过使用0.05 μm VMWP滤器(Millipore)对DNA级H2O透析1小时将pBluescript-3ZFPL的连接反应脱盐。将pBluescript-6ZFP文库的连接反应应用于NucleoSpin Extract II试剂盒，并将DNA洗脱在15 μl去离子水中。将约60 ng脱盐的DNA与50 μl NEB 10-β电感受态大肠杆菌(New England Biolabs)混合，并如制造商建议使用EasyjecT Plus、2.5 kV、25 μF和2 mm电穿孔杯进行电穿孔。对每一文库进行多次电穿孔，并随后直接合并细胞以提高文库大小。热激转化或电穿孔后，向细菌应用SOC培养基，并在37℃和250 rpm下孵育1小时后，将30 μl的SOC培养物用于连续稀释，并铺至含有氨苄青霉素的LB平板上。第二天，测定获得文库克隆的总数。此外，选择每一文库的十个克隆以分离质粒DNA并通过限制酶消化检查插入片段的整合。对这些质粒中的至少三个进行测序以验证文库的多样性。将剩余的SOC培养物转移至100 ml含氨苄青霉素的LB培养基中并在37℃和250 rpm下培养过夜。将那些细胞用于制备每一文库的质粒Midi DNA。
对于酵母单杂交筛选，将六聚锌指蛋白文库转移到相容的猎物载体中。为此目的，通过用XhoI/EcoRI切割载体并插入到退火的寡核苷酸OAN971 和 中来修饰pGAD10 (Clontech)的多克隆位点。将获得的载体pAN1025切割并去磷酸化，6ZFP文库插入片段通过XhoI/SpeI从pBluescript-6ZFPL中释放。如上所述对pBluescript-6ZFP文库完成连接反应和电穿孔入NEB 10-β电感受态大肠杆菌中。
对于灵敏度提高的酵母单杂交筛选，将6ZFP文库转移到另一个相容的猎物载体中。为此目的，将pAN1025的1460bp SphI片段连接入pAN1373（修饰的pRS315）中(Sikorski, R.S.和Hieter, P., 1989, Genetics 122(1), 19-27)，其中用退火的寡核苷酸和代替ApaI, NarI片段。这种修饰导致含有相比于与上文概述的文库克隆方案相容的高拷贝2 μ复制起点的低拷贝ARS/CEN的酵母单杂交载体。
用于组合的分泌型萤光素酶和碱性磷酸酶测定的受体启动子区的克隆
将含有ETRA、ETRB、TLR4或FCER1A启动子区的DNA片段克隆入pAN1485 (NEG-PG04, GeneCopeia)或pAN1486 (EF1a-PG04, GeneCopeia)中，产生含有在受体启动子控制下的分泌型Gaussia萤光素酶和在组成型CMV启动子控制下的分泌型碱性磷酸酶并允许萤光素酶信号对碱性磷酸酶信号归一化的报道基因质粒。详细而言，使用OAN981和从人基因组DNA扩增ETRA启动子，并经SacI/XhoI克隆入pBluescript中，产生pAN1031。将ETRA启动子使用XhoI/Klenow/BamHI从pAN1031中切出，并克隆入HindIII/Klenow/BglII切割的pAN1486中，产生pAN1492。使用和 从人基因组DNA中扩增ETRB启动子，并经SacI/KpnI克隆入pBluescript中，产生pAN1432。将ETRB启动子经StuI/EcoRI从pAN1432中切出，并克隆入用HindIII/Klenow/EcoRI切割的pAN1486中，产生pAN1489。使用 和 从人基因组DNA中扩增TLR4启动子，并经SalI/KpnI克隆入pBluescript中，产生pAN1433。将TLR4启动子用StuI/BamHI从pAN1433中切出，并克隆入HindIII/Klenow/BglII切割的pAN1486中，产生pAN1491。使用 和 从pAN1491中扩增TLR4启动子，并经EcoRV/HindIII克隆入pAN1486中，产生pAN1509。使用OAN1236和 从人基因组DNA中扩增FCER1A启动子，并经SacI/KpnI克隆入pBluescript中，产生pAN1434。将FCER1A启动子经StuI/EcoRI从pAN1434中切出，并经HindIII/Klenow/EcoRI克隆入pAN1486中，产生pAN1490。使用OAN1261和OAN1262 从pAN1490中扩增FCER1A启动子，并经EcoRV/HindIII克隆入pAN1485中，产生pAN1515。
诱饵质粒的克隆
对于每一诱饵质粒，使用侧翼为取自在ETRA、ETRB、TLR4或FCER1A启动子上游和下游的序列的21 bp的18 bp靶位点。包括NcoI位点用于限制性分析。以这样的方式设计寡核苷酸并退火，以产生允许用HindIII/XhoI切割并直接连接入pAbAi (Clontech, )中的5'HindIII和3'XhoI位点（表5）。
表5：用于将靶位点克隆入pAbAi载体中的寡核苷酸
用于哺乳动物转染的人工转录因子的克隆
对于生成DNA片段XbaI-EcoRV-NNNNNN-XhoI-NNNNNN-AgeI-3xmyc-STOP-NotI-EcoRI，用Platinum Pfx DNA聚合酶、 和 将3xmyc标签从pWS250中扩增，用XbaI/EcoRI切割，连接入用XbaI/EcoRI切割的pcDNA3(-)中，产生pAN1109。使用Platinum Pfx DNA聚合酶、 和 从pAN1021中扩增KRAB-NLS。将PCR产物用EcoRV/XhoI切割并连接入用EcoRV/XhoI切割的pAN1109中，产生pAN1110。
用Platinum Pfx DNA聚合酶、 、将ZFP-855A的DNA序列从pAN1021中扩增，用XhoI/AgeI消化并克隆入用XhoI/AgeI切割的pAN1110中，以产生pAN1111。类似地，用 和 从pAN1022中扩增ZFP+74A，克隆入pAN1110中，并产生pAN1112。
使用XhoI/AgeI消化通过合适的6ZFP（通过酵母单杂交鉴定到的）例如通过ZFP-855C代替pAN1111的ZFP-855A，产生pAN1133。
此外，通过在第一DNA合成步骤中退火 、 、 、 使用Platinum Pfx DNA聚合酶生成SID-NLS (SID对应于根据Beerli, R.R.等, 1998, Proc Natl Acad Sci U S A 95, 14628-14633的Mad mSin3相互作用结构域的氨基酸1-36)。将该PCR产物的等分试样作为模板与Platinum Pfx DNA聚合酶、OAN1096和OAN1099用于第二DNA合成步骤。将第二PCR产物用XhoI/EcoRV切割，并用于代替用XhoI/EcoRV切割的pAN1111中的KRAB-NLS。将所获的质粒pAN1208用于在XhoI/AgeI处理后用任何来自Y1H筛选的6ZFP代替ZFP-855A。
此外，根据Gommans, W.M. 等, 2007, Mol Carcinog 46, 391-401按N-末端NLS，随后为6ZFP、GGSGGS (SEQ ID NO: 9)接头序列、人KRAB的氨基酸11-55和C-末端3xmyc标签将蛋白结构域的次序重排列。首先，DNA片段AgeI-EcoRI-NNNNNN-BamHI-3xmyc-STOP-NotI-HindIII用pAN1133作为模板、Platinum Pfx DNA聚合酶、 、 通过PCR生成，用AgeI/HindIII切割，连接入用AgeI/HindIII切割的pAN1109中，产生pAN1183。其次，EcoRV-ATG-NLS-XhoI-XmaI-ZFP-855C-AgeI-GGSGGS接头-EcoRI用pAN1133作为模板、Platinum Pfx DNA聚合酶、 、 通过PCR产生，并使用EcoRI/EcoRV克隆插入到pAN1183中，产生pAN1184。第三，将人KRAB的氨基酸11-55用Platinum Pfx DNA聚合酶、 、 从pAN1133中扩增，用EcoRI/BamHI切割，并连接入用EcoRI/BamHI切割的pAN1184中。将最终的质粒pAN1185通过用XhoI/AgeI切割用于用任何来自Y1H筛选的6ZFP代替ZFP-855C。
对于激活ATF的克隆，通过用EcoRI/BamHI切割并插入退火的OAN1253 (SEQ ID NO: 264)、OAN1254 (SEQ ID NO: 265)、OAN1255 (SEQ ID NO: 266)和OAN1256 (SEQ ID NO: 267)中，将C-末端KRAB结构域用VP64编码序列代替。
改良的酵母单杂交（Y1H）筛选
酵母菌株和培养基
酿酒酵母（Saccharomyces cerevisiae）Y1H Gold购自Clontech，YPD培养基和YPD琼脂购自Carl Roth。合成缺陷型(SD)培养基含有20 g/l葡萄糖、6.8 g/l Na2HPO4·2H2O、9.7 g/l NaH2PO4·2H2O (均来自Carl Roth)、1.4 g/l酵母合成缺陷型培养基添加物、6.7 g/l酵母氮源基础、0.1 g/l L-色氨酸、0.1 g/l L-亮氨酸、0.05 g/l L-腺嘌呤、0.05 g/l L-组氨酸、0.05 g/l尿嘧啶(均来自Sigma-Aldrich)。SD-U培养基含有除了尿嘧啶之外的所有组分，制备不含L-亮氨酸的SD-L。SD琼脂板不含有磷酸钠，但含有16 g/l Bacto琼脂(BD)。金担子素A (AbA)购自Clontech。
诱饵酵母菌株的制备
在20 μl的总体积中用BstBI将约5 μg的每种诱饵质粒线性化，并将一半的反应混合物直接用于酿酒酵母Y1H Gold的热激转化。在转化前一天将酵母细胞用于接种5 ml YPD培养基，并在滚瓶机（roller）上在RT下生长过夜。将一毫升该预培养物用新鲜YPD培养基1：20稀释，并在30℃, 225 rpm下孵育2-3小时。对于每一转化反应，通过离心收获1 OD600，将酵母细胞用1 ml无菌水洗涤一次并用1 ml TE/LiAc (10 mM Tris/HCl, pH 7.5, 1 mM EDTA, 100 mM醋酸锂)洗涤一次。最后，将酵母细胞重悬于50 μl TE/LiAc中，并与50 μg单链鲑鱼精DNA (Sigma-Aldrich)、10 ul的BstBI线性化的诱饵质粒(见上文)和300 μl PEG/TE/LiAc (10 mM Tris/HCl, pH 7.5, 1 mM EDTA, 100 mM醋酸锂, 50% (w/v) PEG 3350)混合。将细胞和DNA在滚瓶机上在RT下孵育20分钟，随后置于42℃水浴中15分钟。最后，通过离心收集酵母细胞，重悬于100 μl无菌水中，并涂布在SD-U琼脂板上。在30℃下孵育3天后，选择来自每一转化反应的生长在SD-U上的8个克隆，以分析它们对金担子素A (AbA)的敏感性。预培养物在滚瓶机上在RT下生长过夜。对于每一培养物，测量OD600，并用无菌水调整为OD600=0.3。从该第一稀释中，用无菌水制备五个额外的1/10稀释步骤。对于每一克隆，将每一稀释步骤的5 μl点在含有SD-U、SD-U 100 ng/ml AbA、SD-U 150 ng/ml AbA和SD-U 200 ng/ml AbA的琼脂板上。在30℃下孵育3天后，选择在SD-U上生长良好并对AbA最敏感的三个克隆用于进一步分析。通过Matchmaker Insert Check PCR Mix 1 (Clontech)按照制造商说明书验证诱饵质粒稳定整合入酵母基因组中。将三个克隆中的一个用于随后的Y1H筛选。
用六聚锌指蛋白文库转化诱饵酵母菌株
将50 μl的酵母诱饵菌株预培养物稀释至100 ml YPD培养基中，并在30℃和225 rpm下孵育直到OD600=1.6-2.0 (大约20小时)。通过在水平转子(5分钟, 1500×g, 4℃)中离心收集细胞。根据Benatuil L.等, 2010, Protein Eng Des Sel 23, 155-159完成电感受态细胞的制备。对于每一转化反应，将400 μl电感受态诱饵酵母细胞与1 μg编码6ZFP文库的猎物质粒混合并在冰上孵育3分钟。将细胞-DNA悬浮液转移至预冷的2 mm电穿孔杯中。电穿孔(EasyjecT Plus电穿孔仪, 2.5 kV和25 μF)后，将酵母细胞转移至8 ml的YPD:1 M山梨醇1:1的混合物中，并在 30℃和225 rpm下孵育90分钟。通过离心收集细胞，并在1 ml的SD-L培养基中重悬。将50 μl的等分试样涂布在含有1000-4000 ng/ml AbA的10 cm SD-L琼脂板上。此外，将50 μl的细胞悬浮液用于制备1/100和1/1000稀释，并将50 μl未稀释的和稀释的细胞铺板至SD-L上。所有平板在30℃下孵育3天。从具有稀释的转化子的平板上计算获得的克隆的总数。尽管用未稀释细胞的SD-L平板表明所有转化子的生长，但如果猎物6ZFP成功结合到它的诱饵靶位点上，则含有AbA的SD-L平板仅产生菌落形成。
阳性相互作用的验证和编码6ZFP的猎物质粒的回收
对于最初分析，从含有最高AbA浓度的SD-L平板上挑出四十个形状良好的菌落，并将酵母细胞在含有3000-4000 ng/ml AbA的SD-L上重新划线两次，以获得单菌落。对于每一克隆，将一个菌落用于接种5 ml SD-L培养基，并将细胞在RT下生长过夜。第二天，用无菌水调节OD600=0.3，制备五个额外的1/10稀释，并将每一稀释步骤的5 μl点在SD-L、SD-L 1000 ng/ml AbA、SD-L 1500 ng/ml AbA、SD-L 2000 ng/ml AbA、SD-L 3000 ng/ml AbA和SD-L 4000 ng/ml AbA的两块平板上。按照它们在高AbA浓度上生长的能力将克隆分级。从最佳生长的克隆中，将5 ml初始SD-L预培养物用于离心下细胞，并将它们在100 μl水或剩余培养基中重悬。加入50 U溶细胞酶(Sigma-Aldrich, L2524)后，将细胞在水平振荡器中在30℃和300 rpm下孵育1小时。将生成的球芽用来自NucleoSpin Plasmid试剂盒的250 μl A1缓冲液稀释，加入一匙尖（one spatula tip）玻璃珠(Sigma-Aldrich, G8772)，并将管通过涡旋剧烈混合20秒。允许玻璃珠沉降，并将250 μl上清液转移至新管中，并用于继续进行标准NucleoSpin Plasmid试剂盒方案。用50 μl洗脱缓冲液洗脱后，通过热激转化或电穿孔将5 μl质粒DNA转化入大肠杆菌DH5α中。 从含氨苄青霉素的LB平板上挑取两个单独的菌落，分离质粒并将文库插入片段测序。将获得结果分析对于每一靶位点的6ZFP之间的共有序列。
细胞培养和转染
HeLa细胞在5% CO2和37℃下生长在添加有4.5 g/l葡萄糖、10%热失活胎牛血清、2 mM L-谷氨酰胺和1 mM丙酮酸钠(均来自Sigma-Aldrich)的Dulbecco's Modified Eagle's Medium (DMEM)中。对于萤光素酶报道基因测定，将7000 HeLa细胞/孔接种入96孔板中。第二天，使用Effectene Transfection Reagent (Qiagen)按照制造商说明书进行共转染。编码人工转录因子和编码萤光素酶的质粒中量制备物（midi preparations）以比例3：1使用。转染后6小时和24小时，每孔用100 μl新鲜的DMEM更换培养基。U937 (Sigma)和KU812F细胞(Sigma)生长在添加有10% FBS、2 mM谷氨酰胺和1 mM丙酮酸钠的RMPI-1640培养基中。使用Cell Line Nucleofector试剂盒C (Amaxa)或Cell Line Nucleofector试剂盒T (Amaxa)按照制造商说明书通过核转染来转染U937和KU812F细胞。RBL-2H3细胞(DSMZ)生长在添加有2 mM谷氨酰胺和1 mM丙酮酸盐的70% MEM/20% RMPI-1640/10%热失活FBS中。使用Cell line nucleofector试剂盒T (Amaxa)核转染RBL-2H3细胞。
按照供应商建议使用平滑肌细胞生长培养基2来生长初级人子宫平滑肌细胞(hUtSMCs, PromoCell)。
组合的萤光素酶/SEAP启动子活性测定
用人工转录因子表达构建体和携带有在ETRA、ETRB、TLR4或FCER1启动子控制下的分泌型Gaussia萤光素酶和在组成型CMV启动子控制下的分泌型碱性磷酸酶(Secrete-Pair Dual Luminescence Assay, GeneCopeia, Rockville, MD)的质粒共转染HeLa或RBL-2H3细胞。转染后两天，收集细胞培养物上清液，并使用Secrete-Pair Dual Luminescence测定(GeneCopoeia)或SEAP报道基因测定(Roche)测量萤光素酶活性和SEAP活性。将用YFP-N1 (Clontech)代替人工转录因子表达构建体的共转染用作对照。将萤光素酶活性对SEAP活性归一化，并表示为对照的百分比。
人子宫平滑肌细胞(hUtSMC)网格收缩测定
将250 μl的无菌牛胶原(3.1 mg/ml; #5005-B Nutacon)与30 μl 10xPBS和22.5 μl 0.1 N NaOH混合以达到pH 7.4。将在200 μl的SMC培养基2中的25000 hUtSMC加入以中和胶原，温柔混合，转移至24孔组织培养板上，并允许在37℃、5% CO2下聚合45分钟。聚合后，加入500 μl的SMC生长培养基2。对于用人工转录因子处理，将1 μM AO74V或作为对照的合适量的缓冲液在聚合后立即加入和在24和48小时后再次加入。聚合后72小时，通过轻柔摇晃或用匙帮助将网格从管壁上脱离下，并加入100 nM的ET-1或缓冲液对照。扫描网格并使用ImageJ软件通过图像分析测定网格面积。
IL-6检测
按照制造商推荐(Amaxa)用TLR4特异性人工转录因子或对照载体的表达质粒核转染1x106 U937细胞。将来自每一核转染的1.25x105个细胞转移至12孔板中，并在用不同LPS浓度刺激8小时前，用100 nM 佛波醇-12-肉豆蔻酸酯-13-乙酸酯(PMA; Sigma)刺激48小时。使用IL-6 ELISA (Orgenium)按照制造商推荐在细胞培养物上清液中分析IL-6的浓度。
IgE结合能力的流式细胞术测定
为了测定IgE与KU812F细胞的结合，将1*106个细胞在2 ml的FACS缓冲液(1x PBS, 2% FBS)中洗涤一次，随后重悬于0.5 ml FACS缓冲液中。将2*105个细胞与10 μg/ml的人IgE (abcam)孵育30分钟，用500 μl FACS缓冲液洗涤一次，随后加入FITC标记的小鼠抗人IgE (5 μg/ml, abcam) 30分钟。将样品在500 μl FACS缓冲液中洗涤一次，并重悬于700 μl FACS缓冲液中。通过流式细胞术(Cyan ADP, Beckman Coulter)分析样品。使用未染色的细胞和仅用FITC标记的小鼠抗人IgE处理的细胞作为对照。
使用MTS测定的细胞增殖的测定
将7000个 HeLa细胞或hUtSMC接种于96孔板的100 μl培养基中，并分别用特异性的人工转录因子或合适的缓冲液对照处理48或72小时。为了测定细胞增殖，按照制造商推荐使用CellTiter 96 Aqueous Non-Radioactive Cell Proliferation Assay (Promega)。将一式三份完成的实验独立重复至少三次。
人工转录因子蛋白的产生
将用给定的人工转录因子的表达质粒转化的大肠杆菌BL21(DE3)生长在添加有100 μM ZnCl2的1L LB培养基中，直至达到0.8-1的OD600，并用1 mM IPTG诱导两小时。通过离心收集细菌，通过超声处理制备细菌裂解物，并纯化包涵体。为此目的，通过离心(5000g, 4℃, 15分钟)收集包涵体，并在20 ml的结合缓冲液(50 mM HEPES, 500 mM NaCl, 10 mM咪唑; pH 7.5)中洗涤三次。将纯化的包涵体在冰上于30 ml结合缓冲液A (50 mM HEPES, 500 mM NaCl, 10 mM咪唑, 6M GuHCl; pH 7.5)中溶解一小时。将溶解的包涵体在4℃和13'000 g下离心40分钟，并通过0.45 μm PVDF滤器过滤。使用His-Trap柱在?ktaprime FPLC (GeHealthcare)上用结合缓冲液A和洗脱缓冲液B(50 mM HEPES, 500 mM NaCl, 500 mM咪唑, 6M GuHCl; pH 7.5)纯化His标签化的人工转录因子。将含有纯化的人工转录因子的级分合并，并在4℃下透析过夜，在人工转录因子含有SID结构域的情况下，对缓冲液S (50 mM Tris-HCl, 500 mM NaCl, 200 mM精氨酸, 100 μM ZnCl2, 5 mM GSH, 0.5 mM GSSG, 50%甘油; pH 7.5)透析，或者对于含有KRAB结构域的人工转录因子，则对缓冲液K (50 mM Tris-HCl, 300 mM NaCl, 500 mM精氨酸, 100 μM ZnCl2, 5 mM GSH, 0.5 mM GSSG, 50%甘油; pH 8.5)透析。透析后，将蛋白样品在4℃下以14'000 rpm离心30分钟，并使用0.22 μm Millex-GV针头式滤器（filter tips）(Millipore)无菌过滤。
统计学分析
使用SPSS (IBM)利用Student's t检验（其中为合适的(Excel, Microsoft Cooperation)或通用的线性单变量模型）完成统计学分析。显示的所有实验为三次独立实验的平均值，并带有代表SEM的误差棒。
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Cys Gly Lys Ser Phe Ser Thr Ser Gly His Leu Val Arg His Gln Arg
115 120 125
Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser
130 135 140
Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly
145 150 155 160
Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser
165 170 175
Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr
180 185 190
Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Glu Leu Val
195 200 205
Arg His Gln Arg Thr His Thr Gly Glu Gln Lys Leu Ile Ser Glu Glu
210 215 220
Asp Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu
225 230 235 240
Ile Ser Glu Glu Asp Leu
245
<210> 14
<211> 279
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 14
Met His His His His His His Gly Tyr Gly Arg Lys Lys Arg Arg Gln
1 5 10 15
Arg Arg Arg Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Pro Trp Asp
20 25 30
Ile Met Ala Ala Ala Val Arg Met Asn Ile Gln Met Leu Leu Glu Ala
35 40 45
Ala Asp Tyr Leu Glu Arg Arg Glu Arg Glu Ala Glu His Gly Tyr Ala
50 55 60
Ser Met Leu Pro Tyr Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro
65 70 75 80
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
85 90 95
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
100 105 110
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
115 120 125
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
130 135 140
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly His Leu Val Arg His Gln
145 150 155 160
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
165 170 175
Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr
180 185 190
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
195 200 205
Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
210 215 220
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Glu Leu
225 230 235 240
Val Arg His Gln Arg Thr His Thr Gly Glu Gln Lys Leu Ile Ser Glu
245 250 255
Glu Asp Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys
260 265 270
Leu Ile Ser Glu Glu Asp Leu
275
<210> 15
<211> 276
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 15
Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro
1 5 10 15
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
20 25 30
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
35 40 45
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu Val Arg His Gln
50 55 60
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
65 70 75 80
Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr
85 90 95
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
100 105 110
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
115 120 125
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
130 135 140
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
145 150 155 160
Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln
165 170 175
Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala
180 185 190
Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu
195 200 205
Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe
210 215 220
Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp
225 230 235 240
Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu
245 250 255
Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser
260 265 270
Glu Glu Asp Leu
275
<210> 16
<211> 276
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 16
Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro
1 5 10 15
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
20 25 30
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
35 40 45
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
50 55 60
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
65 70 75 80
Ser Phe Ser Thr Ser Gly His Leu Val Arg His Gln Arg Thr His Thr
85 90 95
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
100 105 110
Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
115 120 125
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
130 135 140
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
145 150 155 160
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Glu Leu Val Arg His Gln
165 170 175
Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala
180 185 190
Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu
195 200 205
Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe
210 215 220
Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp
225 230 235 240
Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu
245 250 255
Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser
260 265 270
Glu Glu Asp Leu
275
<210> 17
<211> 309
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 17
Met His His His His His His Gly Tyr Gly Arg Lys Lys Arg Arg Gln
1 5 10 15
Arg Arg Arg Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Pro Trp Asp
20 25 30
Ile Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys
35 40 45
Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn
50 55 60
Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys
65 70 75 80
Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His
85 90 95
Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly
100 105 110
Lys Ser Phe Ser Thr Ser Gly His Leu Val Arg His Gln Arg Thr His
115 120 125
Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser
130 135 140
Asp Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys
145 150 155 160
Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser
165 170 175
Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys
180 185 190
Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Glu Leu Val Arg His
195 200 205
Gln Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg
210 215 220
Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala
225 230 235 240
Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp
245 250 255
Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu
260 265 270
Asp Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp
275 280 285
Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile
290 295 300
Ser Glu Glu Asp Leu
305
<210> 18
<211> 246
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 18
Met Ala Ala Ala Val Arg Met Asn Ile Gln Met Leu Leu Glu Ala Ala
1 5 10 15
Asp Tyr Leu Glu Arg Arg Glu Arg Glu Ala Glu His Gly Tyr Ala Ser
20 25 30
Met Leu Pro Tyr Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly
35 40 45
Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Thr
50 55 60
Gly Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr
65 70 75 80
Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu Thr
85 90 95
Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu
100 105 110
Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg
115 120 125
Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser
130 135 140
Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly
145 150 155 160
Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser
165 170 175
Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr
180 185 190
Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Thr
195 200 205
Glu His Gln Arg Thr His Thr Gly Glu Gln Lys Leu Ile Ser Glu Glu
210 215 220
Asp Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu
225 230 235 240
Ile Ser Glu Glu Asp Leu
245
<210> 19
<211> 218
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 19
Met Ala Ala Ala Val Arg Met Asn Ile Gln Met Leu Leu Glu Ala Ala
1 5 10 15
Asp Tyr Leu Glu Arg Arg Glu Arg Glu Ala Glu His Gly Tyr Ala Ser
20 25 30
Met Leu Pro Tyr Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly
35 40 45
Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser
50 55 60
Gly His Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr
65 70 75 80
Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Thr
85 90 95
Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu
100 105 110
Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg
115 120 125
Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser
130 135 140
Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly
145 150 155 160
Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser
165 170 175
Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Gln Lys Leu
180 185 190
Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu
195 200 205
Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu
210 215
<210> 20
<211> 266
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 20
Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro
1 5 10 15
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu
20 25 30
Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
35 40 45
Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly His Leu Thr Glu His Gln
50 55 60
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
65 70 75 80
Ser Phe Ser Thr Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr
85 90 95
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
100 105 110
Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
115 120 125
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
130 135 140
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
145 150 155 160
Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln
165 170 175
Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Arg Thr Leu
180 185 190
Val Thr Phe Lys Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys
195 200 205
Leu Leu Asp Thr Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu
210 215 220
Asn Tyr Lys Asn Leu Val Ser Leu Gly Tyr Gly Ser Glu Gln Lys Leu
225 230 235 240
Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu
245 250 255
Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu
260 265
<210> 21
<211> 266
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 21
Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro
1 5 10 15
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
20 25 30
Ala Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
35 40 45
Glu Cys Gly Lys Ser Phe Ser Arg Ala Asp Asn Leu Thr Glu His Gln
50 55 60
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
65 70 75 80
Ser Phe Ser Ser Arg Arg Thr Cys Arg Ala His Gln Arg Thr His Thr
85 90 95
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
100 105 110
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
115 120 125
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Asn Leu
130 135 140
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
145 150 155 160
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln
165 170 175
Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Arg Thr Leu
180 185 190
Val Thr Phe Lys Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys
195 200 205
Leu Leu Asp Thr Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu
210 215 220
Asn Tyr Lys Asn Leu Val Ser Leu Gly Tyr Gly Ser Glu Gln Lys Leu
225 230 235 240
Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu
245 250 255
Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu
260 265
<210> 22
<211> 299
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 22
Met His His His His His His Gly Tyr Gly Arg Lys Lys Arg Arg Gln
1 5 10 15
Arg Arg Arg Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Pro Trp Asp
20 25 30
Ile Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys
35 40 45
Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His
50 55 60
Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys
65 70 75 80
Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly His Leu Thr Glu His
85 90 95
Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly
100 105 110
Lys Ser Phe Ser Thr Ser Gly Asn Leu Thr Glu His Gln Arg Thr His
115 120 125
Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser
130 135 140
Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys
145 150 155 160
Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn
165 170 175
Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys
180 185 190
Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu Val Arg His
195 200 205
Gln Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Arg Thr
210 215 220
Leu Val Thr Phe Lys Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp
225 230 235 240
Lys Leu Leu Asp Thr Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu
245 250 255
Glu Asn Tyr Lys Asn Leu Val Ser Leu Gly Tyr Gly Ser Glu Gln Lys
260 265 270
Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp
275 280 285
Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu
290 295
<210> 23
<211> 308
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 23
Met Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe
1 5 10 15
Lys Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp
20 25 30
Thr Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys
35 40 45
Asn Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu
50 55 60
Arg Leu Glu Lys Gly Glu Glu Pro Trp Leu Val Glu Arg Glu Ile His
65 70 75 80
Gln Glu Thr His Pro Asp Ser Glu Thr Ala Phe Glu Ile Lys Ser Ser
85 90 95
Val Ser Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys
100 105 110
Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His
115 120 125
Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys
130 135 140
Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu Thr Glu His
145 150 155 160
Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly
165 170 175
Lys Ser Phe Ser Gln Ser Gly Asn Leu Thr Glu His Gln Arg Thr His
180 185 190
Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser
195 200 205
Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys
210 215 220
Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly His
225 230 235 240
Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys
245 250 255
Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His
260 265 270
Gln Arg Thr His Thr Gly Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu
275 280 285
Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser
290 295 300
Glu Glu Asp Leu
305
<210> 24
<211> 341
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 24
Met His His His His His His Gly Tyr Gly Arg Lys Lys Arg Arg Gln
1 5 10 15
Arg Arg Arg Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Pro Trp Asp
20 25 30
Ile Met Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr
35 40 45
Phe Lys Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu
50 55 60
Asp Thr Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr
65 70 75 80
Lys Asn Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile
85 90 95
Leu Arg Leu Glu Lys Gly Glu Glu Pro Trp Leu Val Glu Arg Glu Ile
100 105 110
His Gln Glu Thr His Pro Asp Ser Glu Thr Ala Phe Glu Ile Lys Ser
115 120 125
Ser Val Ser Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu
130 135 140
Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly
145 150 155 160
His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys
165 170 175
Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu Thr Glu
180 185 190
His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys
195 200 205
Gly Lys Ser Phe Ser Gln Ser Gly Asn Leu Thr Glu His Gln Arg Thr
210 215 220
His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe
225 230 235 240
Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu
245 250 255
Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly
260 265 270
His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys
275 280 285
Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg
290 295 300
His Gln Arg Thr His Thr Gly Glu Gln Lys Leu Ile Ser Glu Glu Asp
305 310 315 320
Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile
325 330 335
Ser Glu Glu Asp Leu
340
<210> 25
<211> 6950
<212> DNA
<213> 智人
<400> 25
gtgtctatga atttaactat tctaggtacc tcatctaagt gggataataa aatatctatc 60
tttcctttta tgtctggctt atttcactta acataatatc ttaaacgttc atccatgtgg 120
tagcatatat cattcttttt taaagctgaa taatgttctg tgttatgtac atgtatttat 180
atacatatac atacatgtat ataccacatt ttgtttatcc attcttccac tgatggatat 240
ttggattgtt tccatctttt ggctagtttt caccttttgg cttttctgaa taatgctgct 300
atgaatatgg gtgtacaaat atctgtttga gactctgctt tcaattattt aggtaagtac 360
caaaaagtag aattgctgga tcatagggta attttatgtt taatttttgg agggctgcca 420
tgctgttttc cacaactgct acactatatt ttacattcag actggcaatg cacaagggtt 480
ccaatttctc aacattcttg ccaacacttg ttctttgctg tttttgcttt tgttttacta 540
taatggctat cctaataggt gtgaaggaag aatttttagt aactagtcct acaccacagt 600
gagatcagct gtctcaatag gtgggtcatg atgaatgtgt tctagcaaag actggacaga 660
ttgacatatt cagatatgca ggtgatgcac tgtccaagtg tgtctggcca cagagtgaat 720
aagggctgaa atccagcaca tgtttcacgg gccaagatgt gaactgcctc ttttgggagg 780
aagcagtaag tttttctttc ccgaaaatat tgtcagcttg ccaagccaca tgcccaaagg 840
gtcacctttt tttaatataa acaatggcac ttataaaagc tattagttat tctggttggc 900
tgattctccc tcctagagaa gctgtaagat tagtgaacag ggtaatatct agtctaaccc 960
tactagatga ctattaaggc ctctttcaat ggtggttttc tgtagatctc tttgatggtt 1020
ttacaaaatg gtccctaaat tctttgacac tcctcacact aagggttggg ctctatatac 1080
cctcaccttc aatctgggat ttgtgactgc ttgactaata gaatcaagca gaaatgacac 1140
ggtgccagtt tctgggccca gcccttaaga aactggcagc ttccactttc tgtctctggg 1200
gacattcact cttggatccc cgccaccatg ctgtgaggaa gcccaaacca caagtctcca 1260
caagtctttg tggagagacg cacgtggaga aaaactaaca ctcaacacca agcaagtgaa 1320
ctgtcttgga agtggatcct ccagcccctg ctacatggag caaaaacgag ctgtcccaga 1380
caggctctgc ccaaactgca gactgataag cagagtaaat gagtgttgtt attttaagcc 1440
actacgtttt atagtaattt gtttagctgc agcagatagc cggaacagca tgggatataa 1500
catgaccagt gctccaacct cacacttcta ccatgtgctg actctagcag tgcactgaag 1560
gactccaagg caggccttcc ctgaggagac cccagttttt actcacatgt cacaggcagt 1620
ggcctttgtc cctcatctcc tctcccaccc ccaatttagg ataaagtatc tgccgtgata 1680
aagacgttga gacccacttt ctgtaaggtc ggcttcttca ttgtttgaat ttcttgaggt 1740
ttcacggagc cacgcgctgg aaccttccat agtctctcct gaggctcctt ctttgccctg 1800
ggctggaggt ctgtagccgt gggatgctgg ctacaaggga caagatagaa gcaaaccacc 1860
tgatccagta aactgctgtc cacttcggct cctcaacggc ctctaagctt aagagggagc 1920
acgcaagcca agcaaaggcg gcagggaaga cggagaagaa accacccgtg ggccctggct 1980
ctgtgtccag ttgttccgtc acagatcaaa tctgcctgca ctaagaggat gggttcctct 2040
gcaaggcctt tcggaattct gagtcttgtc tgtcaaactc taccctctct cctccacatc 2100
ccccaccttt tctttcagga aggaaatagt taaaaaagac tcctgccctt cagggcctgg 2160
aagggggcgg cagctttgtg ctttttagtg gccgcgtccc aggatagctg gaaggttagg 2220
acgctcttgc ggtcccagag tggagtggaa ggtctggagc tttgggagga gacggggagg 2280
acagactgga ggcgtgttcc tccggagttt tctttttcgt gcgagccctc gcgcgcgcgt 2340
acagtcatcc cgctggtctg acgattgtgg agaggcggtg gagaggcttc atccatccca 2400
cccggtcgtc gccggggatt ggggtcccag cgagacctcc ccgggagaag cagtgcccag 2460
gaggttttct gaagccgggg aagctgtgca gccgaagccg ccgccgcgcc ggagcccggg 2520
acaccggcca ccctccgcgc cacccaccct cgccggctcc ggcttcctct ggcccaggcg 2580
ccgcgcggac ccggcagctg tctgcgcacg ccgagctcca cggtcggtgc aagtctttct 2640
tatcggggac tgggactggg gcgggtgcgg ggatggcgga gacgctgcct gggcccctcg 2700
gtcgggagaa gacgagagct gggaacgttc tggcccgacc gccctgcagc ttgggcgacc 2760
cgtcgcagca ggggctggca cccacttgcc ccagggcgcg cggggaggcg ggcgccttcc 2820
gtgaggggtg cgctgcggac acgtgtagag ttcggggaac ttcacctctc cacgttaggg 2880
tttaggattc ggagttttag gaagaggaag cccaatagtt ttccctgggt gacccctttt 2940
ccccaggcat gcaagaactt tgaggaacgc caagctctgt ggcttgctcc agcgccttct 3000
tgtgaagttg gctccacagc cttttcttac tggcttgctt ttcctccccc acatctgggt 3060
ctgggtatgc caagggtagc caagggcact cggtgtgagg gactgaggtg tttggggaaa 3120
ccccctgtgt agctagtttt ggaaacctgc ctgtggaggt ctgggcgttt tgttctgtgc 3180
tcgcctttaa aggacacttg ttgcttctca cgtgcagaaa acaagggctg cttctagata 3240
atcaccctct gtttacatca tcctaatggc tttcccctga tagagttggg ggtgggaggc 3300
atattaaact caggcgttgt ctctaaggag agttgctcat ttcttaacca gagagcaaaa 3360
cctcagaaaa cttgtaaaaa gttcctactt ttacttgttt tttcctccac ctgtagttgt 3420
ctatgtgtta gctaattgag aaacaccttg gcagtcattt catgtaaatt gaatgctcta 3480
aactcatgcc attttgataa ggtctgaatt tccaaattac caattacagg tgaactttga 3540
tcctgagagt ctcatatgtt aaatggttaa agtgtaagcg taaggattca gtcctctttc 3600
ttaataatag tttttgcact ccccaaactc tcagattcca aacaggaaga atgctgaatc 3660
tatagaccat gtccttcagg tctgaacctg agatgcttca ctagcgctat gactgattat 3720
gagtgatgat tcaaagcaca gcctgactca gattcctgga aatgagctac agacccacct 3780
tggaagaacc tgcagttgag gggcttcttt catttggttt tgtacttttc agccaatgat 3840
aatatgacat ccacaggcct ttgccaagta tcccttagtt ttgcagctta acatgtgtgg 3900
agagctgata gagaagtcat cagaaatgat ttctgtagtg gtgtcattgt tggaggagga 3960
aggaaggggg ttttaatgat agctggctct catgccttaa gaacccacag cgttggcctt 4020
acctcctggc ataatcagct aaccaagcag accagcatgc taacgttcaa agcaccacag 4080
gacctctgat gagtagagca gtacccaccg tagccaggtc agttcacagg tcagttccca 4140
gccgactgta accggcagag agccattcca tttctgcaaa ggccttctct ctctcatttt 4200
taagaaagct gttattgaag gttccctttc atttgggaag ctggaagacg ttagtcacga 4260
aacagcacaa gatatgaaag tatttcttct cccccctgta tctgtgatcc aaaaaggagg 4320
atgacaagaa gcttttctct tggagtcctg gtgttccctt tggaatggaa atggtgtcat 4380
ttcataggtc tatcaatttt tggccagata ctcactccag attataagaa gcaggaaaaa 4440
actgcaaact gtttataatt aagtctagtc agagctgaca aatacatggc ctgcctctag 4500
ccttgaaaaa ttgatatttt tcccatttaa tcagagtgag tcaagtccta taagcaaaat 4560
tatgtgtcct gccatttctt ggctttgaaa ccttgaccta aatacttctt caagactctg 4620
gtttccttat aagtaaaatg gagatattgc tcctgcctac ctagagtcgt aaagctcaag 4680
gcagataaca gatacatata taataagtta ctatatgtaa atactattat tatttcagag 4740
ttcaggggga aatctctggg gaaggcaaaa gtatccaata ctcgcacttt atatataccc 4800
tatactttct atgcaacttg aataaatctt attttatcca cgtattggcc aaacctaagc 4860
tttactgatt tcccaagata attgtcaaaa ttcctaaaag tggttaacat caaccttgaa 4920
tacactcaga aaaaggattc aattttattt tttacttttt atttatttaa tttttttttt 4980
tttgagacag ggtctcactc tgctgctcag gatggagtgc agtgactcga tcatagctcc 5040
ctgcagcctt aacctcctgg actcaagtga ttctcccatc tcagcctccc aagtagctga 5100
gactacaggc acacgccacc atgcctgtct aatctttttt atgttttgta aagacagggt 5160
ctcactaagt tgcccaggct agtttcaaac tcctgggctc aagtgatcct cctgctttgg 5220
cctcccaaag tgctgagatt atagacatga gccaccacac ccaaccagga ctcaattttt 5280
ttaagattaa attatgacct gggtatatac atcacagaca cgtacacaca ccaccgcata 5340
aatcagatta tgtcttcatt tgaagattca taaaagccta cagaaaagga aatatataaa 5400
atattgaaat aggatgagct atttttaatt atctttgatt attccttatt aaggtctcct 5460
agacttcctg gacaggaagg accacttgct ctctgcaagt gacttaaaaa taaataaata 5520
aataaataaa ccacaccaga tggtctttga aaatgtctaa ccaccctcct gagtctattg 5580
cttcatctaa actctggaga cttctttaaa atattttaca tacaaataaa gcacaagaat 5640
gagagacagg atggtttagt ggttaacagc tccggctcag agttggactg acctgcctta 5700
gaatcgcagt gttgccaccc atcacctgtg gccttgacaa gctgcttcac cctcctgggc 5760
ctctgtttcc ctatcttcaa aattgaaccg tggatctacc ccacaggacc attgtatgga 5820
ttacagggca tcatgcacat atagtactga gtacagtgac tggcataaaa taaatgtagc 5880
tatataggat ggttaaaaga tagcatgggg acaggtgtcc cttggaaacc aaaatactat 5940
gactagcctg gaaaagttca ttactcccat ttcattcatc ggcaaatacc gtattgtgat 6000
gataatttct gagaaatgaa aaagcaaaaa aactcctgta aaaaatttag attttacaca 6060
ttaatcatca aggttttcaa aacaaatgtt tttatcaatt atttcattct taattgaaac 6120
taaaattgtt agatgtgtta atggtgttaa ccagagttct cattaacatt tcataacact 6180
ttagatggct acctgtgctg cacttcatcc ctatgttcat tatttatcat atgcaagaca 6240
gcacacattt tagaagattc aatttttaca ttcgataatt ttattttatg cctcaatacc 6300
ctgtaccctt tactcagcat ataatgcctt ttctctttca ttaaaaaata tgtttgtatc 6360
tcaactcttg aagatttttg taattcaggt tcttatcagc ctgaatgtag agaaagaagt 6420
aactactact taaaggaaaa actgacatag cctcatccat attatgatgt agatataagt 6480
atagctatgt atagagatga gtgagagaga caaagctaga gagataggga gatggagata 6540
gagctagaga tggaaataga gacagagata gatagctata gagacggaaa tggagacaga 6600
gacagagacg gagatgacag agatgatgga gatgatggag atggagatgg agactgacat 6660
agaggagata gagcttgccc taatagagct tcagtttttt tctggttttg ctctgtcata 6720
gggtaacctg atatacatat atcttatcta atcaccaaat atatcctgca ttttaattga 6780
atcaataaat aatcattgac tatgatcttt ttggcaactg ggttttggaa caaaaattat 6840
ttttcctttt gtttcaggtg aaaaaaaagt gaaggtgtaa aagcagcaca agtgcaataa 6900
gagatatttc ctcaaatttg cctcaagatg gaaacccttt gcctcagggc 6950
<210> 26
<211> 1284
<212> DNA
<213> 智人
<400> 26
atggaaaccc tttgcctcag ggcatccttt tggctggcac tggttggatg tgtaatcagt 60
gataatcctg agagatacag cacaaatcta agcaatcatg tggatgattt caccactttt 120
cgtggcacag agctcagctt cctggttacc actcatcaac ccactaattt ggtcctaccc 180
agcaatggct caatgcacaa ctattgccca cagcagacta aaattacttc agctttcaaa 240
tacattaaca ctgtgatatc ttgtactatt ttcatcgtgg gaatggtggg gaatgcaact 300
ctgctcagga tcatttacca gaacaaatgt atgaggaatg gccccaacgc gctgatagcc 360
agtcttgccc ttggagacct tatctatgtg gtcattgatc tccctatcaa tgtatttaag 420
ctgctggctg ggcgctggcc ttttgatcac aatgactttg gcgtatttct ttgcaagctg 480
ttcccctttt tgcagaagtc ctcggtgggg atcaccgtcc tcaacctctg cgctcttagt 540
gttgacaggt acagagcagt tgcctcctgg agtcgtgttc agggaattgg gattcctttg 600
gtaactgcca ttgaaattgt ctccatctgg atcctgtcct ttatcctggc cattcctgaa 660
gcgattggct tcgtcatggt accctttgaa tataggggtg aacagcataa aacctgtatg 720
ctcaatgcca catcaaaatt catggagttc taccaagatg taaaggactg gtggctcttc 780
gggttctatt tctgtatgcc cttggtgtgc actgcgatct tctacaccct catgacttgt 840
gagatgttga acagaaggaa tggcagcttg agaattgccc tcagtgaaca tcttaagcag 900
cgtcgagaag tggcaaaaac agttttctgc ttggttgtaa tttttgctct ttgctggttc 960
cctcttcatt taagccgtat attgaagaaa actgtgtata acgagatgga caagaaccga 1020
tgtgaattac ttagtttctt actgctcatg gattacatcg gtattaactt ggcaaccatg 1080
aattcatgta taaaccccat agctctgtat tttgtgagca agaaatttaa aaattgtttc 1140
cagtcatgcc tctgctgctg ctgttaccag tccaaaagtc tgatgacctc ggtccccatg 1200
aacggaacaa gcatccagtg gaagaaccac gatcaaaaca accacaacac agaccggagc 1260
agccataagg acagcatgaa ctga 1284
<210> 27
<211> 1180
<212> DNA
<213> 智人
<400> 27
tgtccccgga cgaggactgc ccccctccct cgggcaacta ctactgatgc tgtccaggca 60
tcgcccaagg ggaaaggttg cagcggggtc ggaaggcgcg ggaggagtct ggcggtgatt 120
gatgggaagg gatgaatgaa taaaagtact tgtctgatgg cagcagagac cccgagcaaa 180
cggtggaggc tacactgtct ggcattctcg cagcgtttcg tcagagccgg acccgcctgc 240
agctcaaggg aggcgtgctc ctctcccaga gcaggctgga acccagctgg gttccgcctc 300
ccgggaaggt ggtctccatt cgtcgctctg catctggttt gtcagatccg agaggtaaac 360
attcgggctt ggtgttgaat taaaatcatt gattgaacct tattctgggg cttcggtttg 420
gcttactagt ttgggatttt aaaaaaataa aaattaagcc tatagagagg gcaaattaaa 480
attaggttgg gtaaaggaag gagcgcgagt gtttgaagcc gtttggaggg aacagcggtt 540
tccaagttcc tgctgacttg agaagtctct gcgggtttcc gaatctccgg cgcactcctg 600
ggcgcgctgc gggagctgta gctcagccag ccagggagta gcggctttca tccgccggga 660
ggagtctttc gagttcaatc gcggggtata gaggttcccc tgcggggcaa aatgcagagc 720
ttgacacaag cccttggcct ctaggtgcct taattccgcg gttcccacgc acgcttaact 780
aagacgtgtc tgtattcctc ccgttacgtg aaagagttcg gagctttgcc tgggaccccc 840
atcattccct ccctggcaca ccccttccag aacgccccgc cccactgcat attatttacc 900
cctcctggcc acgcggggga agaaaaacag ctgagagggc atcaggaagg agtttcgacc 960
cgcgctggcg agtcatgagc gccaagtttc ccactggcgc gcaaacttga gttacttttg 1020
agcgtggata ctggcgaaga ggctgcgggc ggtattagcg tttgcagcga cttggctcgg 1080
gcagctgacc caagtgtcct gtcttccttc ctctgcttgt ctctaggctc tgaaactgcg 1140
gagcggccac cggacgcctt ctggagcagg tagcagcatg 1180
<210> 28
<211> 644
<212> DNA
<213> 智人
<400> 28
ccttgagttc agactggaag cctctagaat tgtgagaaaa tgaatgtctg ttgtttaagc 60
cacccagtct gtggtatttc cttatggcag ccccagcaaa ctaatacaaa tagtgtttcc 120
acagctgaaa caaaattgga aaatcaccgt catcctagag agttacaagg gctattttaa 180
tagaacctga ttgttttcct aaattcacca agcccaggca gaggtcagat gactaattgg 240
gataaaagcc aactagcttc ctcttgctgt ttctttagcc actggtctgc aggcgttttc 300
ttcttctaac ttcctctcct gtgacaaaag agataactat tagagaaaca aaagtccaga 360
atgctaaggt tgccgctttc acttcctctc accctttagc ccagaactgc tttgaataca 420
ccaattgctg tggggcggct cgaggaagag aagacaccag tgcctcagaa actgctcggt 480
cagacggtga tagcgagcca cgcattcaca gggccactgc tgctcacaga agcagtgagg 540
atgatgccag gatgatgtct gcctcgcgcc tggctgggac tctgatccca gccatggcct 600
tcctctcctg cgtgagacca gaaagctggg agccctgcgt ggag 644
<210> 29
<211> 220
<212> DNA
<213> 智人
<400> 29
taagtgggta aatattaaat tgcccagttg ggcaccatcc tgaatattat ctctaaagaa 60
agaagcaaaa ccaggcacag ctgatgggtt aaccagatat gatacagaaa acatttcctt 120
ctgctttttg gttttaagcc tatatttgaa gccttagatc tctccagcac agtaagcacc 180
aggagtccat gaagaagatg gctcctgcca tggaatcccc 220
<210> 30
<211> 18
<212> DNA
<213> 智人
<400> 30
tcctccagcc cctgctac 18
<210> 31
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 31
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu
35 40 45
Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 32
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 32
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu
35 40 45
Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly His Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 33
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 33
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Ala Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 34
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 34
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu
35 40 45
Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly His Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 35
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 35
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly His Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly Ala Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 36
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 36
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly His Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 37
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 37
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu
35 40 45
Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Asp Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 38
<211> 15
<212> DNA
<213> 智人
<400> 38
ctcctctccc acccc 15
<210> 39
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 39
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 40
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 40
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu
35 40 45
Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asp Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 41
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 41
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu
35 40 45
Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 42
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 42
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 43
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 43
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly Ala Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu
35 40 45
Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 44
<211> 15
<212> DNA
<213> 智人
<400> 44
aaggtcggct tcttc 15
<210> 45
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 45
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 46
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 46
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Thr Gly Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 47
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 47
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser
1 5 10 15
Arg Arg Thr Cys Arg Ala His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 48
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 48
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser
1 5 10 15
Lys Lys His Leu Ala Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 49
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 49
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Thr Gly Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
35 40 45
Ala Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 50
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 50
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 51
<211> 15
<212> DNA
<213> 智人
<400> 51
cggagccacg cgctg 15
<210> 52
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 52
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Gly His Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 53
<211> 18
<212> DNA
<213> 智人
<400> 53
cggctcctca acggcctc 18
<210> 54
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 54
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Lys Asn Ser Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
115 120 125
Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 55
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 55
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 56
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 56
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Asn Ser Thr Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 57
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 57
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser His Ser Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 58
<211> 18
<212> DNA
<213> 智人
<400> 58
ccacccgtgg gccctggc 18
<210> 59
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 59
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly His Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Thr Gly Ala Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 60
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 60
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ala Asp Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 61
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 61
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Thr Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Thr Gly Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 62
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 62
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Asn Asp Thr Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys
145 150 155 160
Arg Ala His Gln Arg Thr His Thr
165
<210> 63
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 63
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly His Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys
145 150 155 160
Arg Ala His Gln Arg Thr His Thr
165
<210> 64
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 64
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly His Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys
145 150 155 160
Arg Ala His Gln Arg Thr His Thr
165
<210> 65
<211> 18
<212> DNA
<213> 智人
<400> 65
ctcctccaca tcccccac 18
<210> 66
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 66
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Ala Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 67
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 67
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 68
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 68
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Cys Arg Asp Leu Ala Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly His Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 69
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 69
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asp Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Ala Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 70
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 70
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 71
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 71
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
35 40 45
Ala Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 72
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 72
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 73
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 73
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Glu Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 74
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 74
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
35 40 45
Ala Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 75
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 75
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
115 120 125
Pro Gly Ala Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 76
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 76
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Ala Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 77
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 77
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asp Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 78
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 78
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asp Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 79
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 79
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Ala Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 80
<211> 140
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 80
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
115 120 125
Pro Gly Ala Leu Val Arg His Gln Arg Thr His Thr
130 135 140
<210> 81
<211> 18
<212> DNA
<213> 智人
<400> 81
ggcctggaag ggggcggc 18
<210> 82
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 82
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 83
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 83
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu
35 40 45
Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 84
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 84
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly Ala Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 85
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 85
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ala Asp Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 86
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 86
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu
35 40 45
Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Asn Asp Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 87
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 87
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Thr Gly Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 88
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 88
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu
35 40 45
Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser
115 120 125
Lys Lys His Leu Ala Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 89
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 89
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 90
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 90
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Asn Asp Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 91
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 91
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Cys Arg Asp Leu Ala Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 92
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 92
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 93
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 93
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Glu Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Asp Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Ala Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 94
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 94
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly Ala Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu
35 40 45
Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Asn Asp Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 95
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 95
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu
35 40 45
Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu
145 150 155 160
Glu Arg His Gln Arg Thr His Thr
165
<210> 96
<211> 18
<212> DNA
<213> 智人
<400> 96
ggaggagacg gggaggac 18
<210> 97
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 97
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu
145 150 155 160
Glu Arg His Gln Arg Thr His Thr
165
<210> 98
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 98
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 99
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 99
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Ala Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 100
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 100
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 101
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 101
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 102
<211> 164
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 102
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ile
1 5 10 15
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
20 25 30
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln
35 40 45
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
50 55 60
Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln Arg Thr His Thr
65 70 75 80
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
85 90 95
Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
100 105 110
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
115 120 125
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
130 135 140
Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln
145 150 155 160
Arg Thr His Thr
<210> 103
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 103
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly His Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
115 120 125
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 104
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 104
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly His Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 105
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 105
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Ala Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 106
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 106
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 107
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 107
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly Ala Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 108
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 108
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 109
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 109
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly Ala Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Glu Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
115 120 125
Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 110
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Ala Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu
145 150 155 160
Glu Arg His Gln Arg Thr His Thr
165
<210> 111
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 111
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 112
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 112
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu
145 150 155 160
Glu Arg His Gln Arg Thr His Thr
165
<210> 113
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 113
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 114
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 114
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
115 120 125
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 115
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 115
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly His Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 116
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 116
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly His Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
115 120 125
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 117
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 117
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
115 120 125
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Ala Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 118
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 118
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly His Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Glu Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 119
<211> 18
<212> DNA
<213> 智人
<400> 119
ctcgggcaac tactactg 18
<210> 120
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 120
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Asn Asp Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys
35 40 45
Arg Ala His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Asn Ser Thr Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Ser Lys Lys His Leu Ala Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 121
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 121
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Asn Asp Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 122
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 122
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser
1 5 10 15
Arg Arg Thr Cys Arg Ala His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 123
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 123
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser
1 5 10 15
Lys Lys His Leu Ala Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 124
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 124
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys
35 40 45
Arg Ala His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Lys Asn Ser Leu Thr Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
115 120 125
Cys Arg Asp Leu Ala Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Lys Lys His Leu
145 150 155 160
Ala Glu His Gln Arg Thr His Thr
165
<210> 125
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 125
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys Arg Ala His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 126
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 126
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Thr Gly Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Ser Lys Lys His Leu Ala Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 127
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 127
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Asn Asp Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Thr Gly Ala Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Thr Gly Ala Leu Thr Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 128
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 128
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Lys Asn Ser Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys
35 40 45
Arg Ala His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 129
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 129
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser His Thr Gly His Leu
35 40 45
Leu Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 130
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 130
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Asn Asp Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 131
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 131
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Thr Gly Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys
35 40 45
Arg Ala His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 132
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 132
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Lys Asn Ser Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys
35 40 45
Arg Ala His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 133
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 133
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Thr Gly Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 134
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 134
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser
1 5 10 15
Lys Lys His Leu Ala Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
115 120 125
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 135
<211> 140
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 135
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Gly His Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr
130 135 140
<210> 136
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 136
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Thr Gly Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys Arg Ala His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Ser Arg Arg Thr Cys Arg Ala His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 137
<211> 18
<212> DNA
<213> 智人
<400> 137
gaggttcccc tgcggggc 18
<210> 138
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 138
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Ser Arg Arg Thr Cys Arg Ala His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 139
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 139
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Ser Lys Lys His Leu Ala Glu His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 140
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 140
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Ser Arg Arg Thr Cys Arg Ala His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 141
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 141
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Thr Gly Ala Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Ser Arg Arg Thr Cys Arg Ala His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 142
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 142
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Ser Arg Arg Thr Cys Arg Ala His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 143
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 143
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Ser Arg Arg Thr Cys Arg Ala His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 144
<211> 18
<212> DNA
<213> 智人
<400> 144
caccaagccc aggcagag 18
<210> 145
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 145
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ala Asp Asn Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 146
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 146
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu
35 40 45
Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ala Asp Asn Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Lys Asn Ser Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ala Asp Asn Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 147
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 147
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ala Asp Asn Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Thr Gly Ala Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 148
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 148
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu
35 40 45
Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 149
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 149
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu
35 40 45
Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ala Asp Asn Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Lys Asn Ser Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys
145 150 155 160
Arg Ala His Gln Arg Thr His Thr
165
<210> 150
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 150
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu
35 40 45
Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ala Asp Asn Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asn Leu
145 150 155 160
Thr Glu His Gln Arg Thr His Thr
165
<210> 151
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 151
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu
35 40 45
Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ala Asp Asn Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Lys Asn Ser Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys
145 150 155 160
Arg Ala His Gln Arg Thr His Thr
165
<210> 152
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 152
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu
35 40 45
Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ala Asp Asn Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys
145 150 155 160
Arg Ala His Gln Arg Thr His Thr
165
<210> 153
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 153
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Glu Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys
145 150 155 160
Arg Ala His Gln Arg Thr His Thr
165
<210> 154
<211> 18
<212> DNA
<213> 智人
<400> 154
gctgtggggc ggctcgag 18
<210> 155
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 155
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Thr Gly Ala Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys Arg Ala His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 156
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 156
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly His Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 157
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 157
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Thr Gly Ala Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Ser Lys Lys His Leu Ala Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Glu Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
115 120 125
Cys Arg Asp Leu Ala Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 158
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 158
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Glu Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 159
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 159
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Cys Arg Asp Leu Ala Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ala Asp Asn Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys Arg Ala His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 160
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 160
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser His Ser Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 161
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 161
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 162
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 162
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser His Ser Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 163
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 163
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Thr Gly Ala Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 164
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 164
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Cys Arg Asp Leu Ala Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Asn Ser Thr Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 165
<211> 18
<212> DNA
<213> 智人
<400> 165
atggccttcc tctcctgc 18
<210> 166
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 166
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Asn Ser Thr Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 167
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 167
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser
1 5 10 15
Lys Lys His Leu Ala Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 168
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 168
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu
35 40 45
Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
115 120 125
Pro Gly Ala Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 169
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 169
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser
1 5 10 15
Lys Lys His Leu Ala Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 170
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 170
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser
1 5 10 15
Lys Lys His Leu Ala Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
115 120 125
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu
145 150 155 160
Arg Arg His Gln Arg Thr His Thr
165
<210> 171
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 171
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
115 120 125
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu
145 150 155 160
Arg Arg His Gln Arg Thr His Thr
165
<210> 172
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 172
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Asn Asp Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
115 120 125
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 173
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 173
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser
1 5 10 15
Arg Arg Thr Cys Arg Ala His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly His Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu
145 150 155 160
Ala Arg His Gln Arg Thr His Thr
165
<210> 174
<211> 18
<212> DNA
<213> 智人
<400> 174
gcccagttgg gcaccatc 18
<210> 175
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 175
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asn Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 176
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 176
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asn Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 177
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 177
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly His Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 178
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 178
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly His Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 179
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 179
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly His Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
115 120 125
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 180
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 180
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asn Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 181
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 181
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu
35 40 45
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly His Leu Thr Glu His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 182
<211> 18
<212> DNA
<213> 智人
<400> 182
gtccatgaag aagatggc 18
<210> 183
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 183
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 184
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 184
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
1 5 10 15
Ser Gly Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 185
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 185
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 186
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 186
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 187
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 187
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
1 5 10 15
Ser Asp Asp Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr
115 120 125
Thr Gly Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 188
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 188
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 189
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 189
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ala Asp Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 190
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 190
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp
1 5 10 15
Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Asn Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 191
<211> 168
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 191
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln
1 5 10 15
Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
20 25 30
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu
35 40 45
Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
50 55 60
Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln
65 70 75 80
Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys
85 90 95
Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg
115 120 125
Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu
145 150 155 160
Val Arg His Gln Arg Thr His Thr
165
<210> 192
<211> 12
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 192
Pro Val Arg Arg Pro Arg Arg Arg Arg Arg Arg Lys
1 5 10
<210> 193
<211> 12
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 193
Thr His Arg Leu Pro Arg Arg Arg Arg Arg Arg Lys
1 5 10
<210> 194
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 194
Arg Arg Arg Arg Arg Arg Arg Arg Arg
1 5
<210> 195
<211> 9
<212> PRT
<213> Influenza A virus
<400> 195
Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
1 5
<210> 196
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 196
Pro Lys Lys Lys Arg Lys Val
1 5
<210> 197
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> 合成构建体
<400> 197
Tyr Lys Asp Asp Asp Asp Lys
1 5
<210> 198
<211> 57
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 198
tcgacaggcc caggcggccc tcgaggatat catgatgact agtggccagg ccggccc 57
<210> 199
<211> 57
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 199
aattgggccg gcctggccac tagtcatcat gatatcctcg agggccgcct gggcctg 57
<210> 200
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 200
cgccgcatgc attcatgcag gcc 23
<210> 201
<211> 17
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 201
tgcatgaatg catgcgg 17
<210> 202
<211> 37
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 202
aatcgcgagc tccttaagaa actggcagct tccactt 37
<210> 203
<211> 31
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 203
aatcgcctcg agctgccggg tccgcgcggc g 31
<210> 204
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 204
gctagctgtc gacacatggt gcgtgataac ttgccc 36
<210> 205
<211> 41
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 205
gctagctggt accaggcctg ctgctacctg ctccagaagg c 41
<210> 206
<211> 40
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 206
gctagctgtc gacataagcc agtgacaaaa agatacatac 40
<210> 207
<211> 44
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 207
gctagctggt accaggcctt atttgatctc tgtggcttct tgag 44
<210> 208
<211> 37
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 208
ctagctgata tcagcttagc ggtttacatg acttgac 37
<210> 209
<211> 34
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 209
ctagctaagc ttcacgcagg agaggaaggc catg 34
<210> 210
<211> 42
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 210
gctagctgtc gacttaaatt cctatttatt aaccttttta gc 42
<210> 211
<211> 42
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 211
gctagctggt accaggcctg tcaccaccca cagtaaaggt tc 42
<210> 212
<211> 37
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 212
ctagctgata tcgctagcca tgctcctgaa tatgtat 37
<210> 213
<211> 43
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 213
ctagctaagc ttggcaggag ccctcttctt catggactcc tgg 43
<210> 214
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 214
agcttgtgaa ctgtcttgga agtggatcct ccagcccctg ctacatggag caaaaacgag 60
ctgtcccatg gc 72
<210> 215
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 215
tcgagccatg ggacagctcg tttttgctcc atgtagcagg ggctggagga tccacttcca 60
agacagttca ca 72
<210> 216
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 216
agcttaggca gtggcctttg tccctcatct cctctcccac ccccaattta ggataaagta 60
tctgcccatg gc 72
<210> 217
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 217
tcgagccatg ggcagatact ttatcctaaa ttgggggtgg gagaggagat gagggacaaa 60
ggccactgcc ta 72
<210> 218
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 218
agcttagacg ttgagaccca ctttctgtaa ggtcggcttc ttcattgttt gaatttcttg 60
aggttccatg gc 72
<210> 219
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 219
tcgagccatg gaacctcaag aaattcaaac aatgaagaag ccgaccttac agaaagtggg 60
tctcaacgtc ta 72
<210> 220
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 220
agcttattgt ttgaatttct tgaggtttca cggagccacg cgctggaacc ttccatagtc 60
tctccccatg gc 72
<210> 221
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 221
tcgagccatg gggagagact atggaaggtt ccagcgcgtg gctccgtgaa acctcaagaa 60
attcaaacaa ta 72
<210> 222
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 222
agcttaaaaa agactcctgc ccttcagggc ctggaagggg gcggcagctt tgtgcttttt 60
agtggccatg gc 72
<210> 223
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 223
tcgagccatg gccactaaaa agcacaaagc tgccgccccc ttccaggccc tgaagggcag 60
gagtcttttt ta 72
<210> 224
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 224
agcttgcgtg ctccctctta agtttagagg ccgttgagga gccgaagtgg acagcagttt 60
actggccatg gc 72
<210> 225
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 225
tcgagccatg gccagtaaac tgctgtccac ttcggctcct caacggcctc taaacttaag 60
agggagcacg ca 72
<210> 226
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 226
agcttggcag ggaagacgga gaagaaacca cccgtgggcc ctggctctgt gtccagttgt 60
tccgtccatg gc 72
<210> 227
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 227
tcgagccatg gacggaacaa ctggacacag agccagggcc cacgggtggt ttcttctccg 60
tcttccctgc ca 72
<210> 228
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 228
agcttgtctg tcaaactcta ccctctctcc tccacatccc ccaccttttc tttcaggaag 60
gaaatccatg gc 72
<210> 229
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 229
tcgagccatg gatttccttc ctgaaagaaa aggtggggga tgtggaggag agagggtaga 60
gtttgacaga ca 72
<210> 230
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 230
agcttagtgg aaggtctgga gctttgggag gagacgggga ggacagactg gaggcgtgtt 60
cctccccatg gc 72
<210> 231
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 231
tcgagccatg gggaggaaca cgcctccagt ctgtcctccc cgtctcctcc caaagctcca 60
gaccttccac ta 72
<210> 232
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 232
agcttggacg aggactgccc ccctccctcg ggcaactact actgatgctg tccaggcatc 60
gcccaccatg gc 72
<210> 233
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 233
tcgagccatg gtgggcgatg cctggacagc atcagtagta gttgcccgag ggaggggggc 60
agtcctcgtc ca 72
<210> 234
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 234
agcttcgagt tcaatcgcgg ggtatagagg ttcccctgcg gggcaaaatg cagagcttga 60
cacaaccatg gc 72
<210> 235
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 235
tcgagccatg gttgtgtcaa gctctgcatt ttgccccgca ggggaacctc tataccccgc 60
gattgaactc ga 72
<210> 236
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 236
agcttacctg attgttttcc taaattcacc aagcccaggc agaggtcaga tgactaattg 60
ggataccatg gc 72
<210> 237
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 237
tcgagccatg gtatcccaat tagtcatctg acctctgcct gggcttggtg aatttaggaa 60
aacaatcagg ta 72
<210> 238
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 238
agcttactgc tttgaataca ccaattgctg tggggcggct cgaggaagag aagacaccag 60
tgcctccatg gc 72
<210> 239
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 239
tcgagccatg gaggcactgg tgtcttctct tcctcgagcc gccccacagc aattggtgta 60
ttcaaagcag ta 72
<210> 240
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 240
agcttgctgg gactctgatc ccagccatgg ccttcctctc ctgcgtgaga ccagaaagct 60
gggagccatg gc 72
<210> 241
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 241
tcgagccatg gctcccagct ttctggtctc acgcaggaga ggaaggccat ggctgggatc 60
agagtcccag ca 72
<210> 242
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 242
agctttaagt gggtaaatat taaattgccc agttgggcac catcctgaat attatctcta 60
aagaaccatg gc 72
<210> 243
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 243
tcgagccatg gttctttaga gataatattc aggatggtgc ccaactgggc aatttaatat 60
ttacccactt aa 72
<210> 244
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 244
agcttccagc acagtaagca ccaggagtcc atgaagaaga tggctcctgc catggaatcc 60
cctacccatg gc 72
<210> 245
<211> 72
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 245
tcgagccatg ggtaggggat tccatggcag gagccatctt cttcatggac tcctggtgct 60
tactgtgctg ga 72
<210> 246
<211> 61
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 246
aatcgctcta gagatatcat atatctcgag atatataccg gtgagcagaa actcatctct 60
g 61
<210> 247
<211> 40
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 247
gcgattgaat tcgcggccgc ttacagatct tcctcagaga 40
<210> 248
<211> 31
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 248
aatcgcgata tcatggatgc taagtccctg a 31
<210> 249
<211> 32
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 249
gcgattctcg agccccactt tacgtttctt tt 32
<210> 250
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 250
aatcgcctcg agcccgggcc gggtgaaaag ccctat 36
<210> 251
<211> 29
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 251
gcgattaccg gtctgtgctg atgagcccc 29
<210> 252
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 252
aatcgcctcg agcccgggcc aggcgaaaag ccctac 36
<210> 253
<211> 30
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 253
gcgattaccg gtctgtgctg aactaccgcc 30
<210> 254
<211> 53
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 254
aatcgcgata tcatggcggc ggcggttcgg atgaacatcc agatgctgct gga 53
<210> 255
<211> 54
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 255
atccagatgc tgctggaggc ggccgactat ctggagcggc gggagagaga agct 54
<210> 256
<211> 47
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 256
ggtatggtaa catggaggca taaccatgtt cagcttctct ctcccgc 47
<210> 257
<211> 53
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 257
gcgattctcg agccccactt tacgtttctt tttcgggtat ggtaacatgg agg 53
<210> 258
<211> 48
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 258
gcgattaccg gtgaattcat atatggatcc gagcagaaac tcatctct 48
<210> 259
<211> 40
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 259
gcgattaagc ttgcggccgc ttacagatct tcctcagaga 40
<210> 260
<211> 34
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 260
gcgattgata tcatgccgaa aaagaaacgt aaag 34
<210> 261
<211> 47
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 261
gcgattgaat tcgctgccgc cgctgccgcc accggtatga gtcctct 47
<210> 262
<211> 28
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 262
gcgattgaat tccgcacact ggttacct 28
<210> 263
<211> 28
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 263
gcgattggat ccatagccca ggctaacc 28
<210> 264
<211> 90
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 264
aattcgggcg cgccgacgcg ctggacgatt tcgatctcga catgctgggt tctgatgccc 60
tcgatgactt tgacctggat atgttgggaa 90
<210> 265
<211> 91
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 265
gtcgcttccc aacatatcca ggtcaaagtc atcgagggca tcagaaccca gcatgtcgag 60
atcgaaatcg tccagcgcgt cggcgcgccc g 91
<210> 266
<211> 81
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 266
gcgacgcatt ggatgacttt gatctggaca tgctcggctc cgatgctctg gacgatttcg 60
atctcgatat gttaattaac g 81
<210> 267
<211> 80
<212> DNA
<213> 人工序列
<220>
<223> 合成构建体
<400> 267
gatccgttaa ttaacatatc gagatcgaaa tcgtccagag catcggagcc gagcatgtcc 60
agatcaaagt catccaatgc 80
Claims (28)
1.人工转录因子，其包含融合至抑制或激活蛋白结构域、核定位序列和蛋白质转导结构域的特异性靶向受体基因启动子的多指锌指蛋白。
2.人工转录因子，其中受体基因启动子是内皮素受体A启动子。
3.人工转录因子，其中受体基因启动子是内皮素受体B启动子。
4.人工转录因子，其中受体基因启动子是Toll样受体4启动子。
5.人工转录因子，其中受体基因启动子是FCER1A启动子。
6.权利要求1、2、3、4或5的人工转录因子，其包含六聚锌指蛋白。
7.权利要求2、3、4或5的人工转录因子，其包含具有选自以下的蛋白序列的锌指蛋白：SEQ ID NO:31至SEQ ID NO:37、SEQ ID NO:39至SEQ ID NO:43、SEQ ID NO:45至SEQ ID NO:50、SEQ ID NO:52、SEQ ID NO:54至SEQ ID NO:57、SEQ ID NO:59至SEQ ID NO:64、SEQ ID NO:66至SEQ ID NO:80、SEQ ID NO:82至SEQ ID NO:95、SEQ ID NO:97至SEQ ID NO:118、SEQ ID NO:120至SEQ ID NO:136、SEQ ID NO:138至SEQ ID NO:143、SEQ ID NO:145至SEQ ID NO:153、SEQ ID NO:155至SEQ ID NO:164、SEQ ID NO:166至SEQ ID NO:173、SEQ ID NO:175至SEQ ID NO:181、和SEQ ID NO:183至SEQ ID NO:191。
8.权利要求2、3、4或5的人工转录因子，其包含具有选自以下蛋白序列的锌指蛋白：SEQ ID NO 56、83、85、101、114、118、127、133、140、142、146、147、156、159、175、和181。
9.权利要求2、3、4或5的人工转录因子，其包含SEQ ID NO 118、133、156、或175的锌指蛋白。
10.权利要求1-9中任一项的人工转录因子，其中所述锌指蛋白融合至抑制蛋白结合域。
11.权利要求10的人工转录因子，其中所述抑制蛋白结构域是SEQ ID NO: 1的N-末端KRAB、SEQ ID NO: 2的C-末端的KRAB、SEQ ID NO: 3的SID、或SEQ ID NO: 4的ERD。
12.权利要求1-9中任一项的人工转录因子，其中所述锌指蛋白融合至激活蛋白结合域。
13.权利要求12的人工转录因子，其中所述激活蛋白结构域是SEQ ID NO: 5的VP16或SEQ ID NO: 6的VP64。
14.权利要求1-13中任一项的人工转录因子，其中所述核定位序列是含有K-K/R-X-K/R共有序列的碱性氨基酸簇或SEQ ID NO: 196的SV40 NLS。
15.权利要求1-14中任一项的人工转录因子，其中所述蛋白质转导结构域是SEQ ID NO: 7的HIV衍生的TAT肽、HSV-1 VP22肽、SEQ ID NO: 192的合成肽mT02、SEQ ID NO: 193的合成肽mT03、SEQ ID NO: 194的R9肽、ANTP结构域、或保护性抗原/致死因子N末端PTD。
16.人工转录因子，其包含融合至抑制或激活蛋白结构域和核定位序列的特异性靶向内皮素受体A启动子的多指锌指蛋白。
17.人工转录因子，其包含融合至抑制或激活蛋白结构域和核定位序列的特异性靶向内皮素受体B启动子的多指锌指蛋白。
18.人工转录因子，其包含融合至抑制或激活蛋白结构域和核定位序列的特异性靶向Toll样受体4启动子的多指锌指蛋白。
19.人工转录因子，其包含融合至抑制或激活蛋白结构域和核定位序列的特异性靶向FCER1A启动子的多指锌指蛋白。
20.药物组合物，其包含权利要求1-19的人工转录因子。
21.权利要求1-19的人工转录因子，其用于调节细胞对外界刺激和对其他可溶性信号分子的反应。
22.权利要求1-19的人工转录因子，其用于治疗由特异性效应物与受体的结合所调节的疾病，为此所述多指锌指蛋白特异性靶向所述受体基因启动子。
23.权利要求2或6-16的人工转录因子，其用于影响对内皮素的细胞应答，以降低或提高内皮素受体A水平，并且用于治疗由内皮素调节的疾病。
24.权利要求3、6-15或17的人工转录因子，其用于影响对内皮素的细胞应答，以降低或提高内皮素受体B水平，并且用于治疗由内皮素调节的疾病。
25.权利要求4、6-15或18的人工转录因子，其用于影响对脂多糖的细胞应答，以降低或提高Toll样受体4水平，并且用于治疗由脂多糖调节的疾病。
26.权利要求5-15或19的人工转录因子，其用于影响对IgE的细胞应答，以降低或提高IgE受体水平，并且用于治疗由IgE调节的疾病。
27.治疗疾病的方法，其包括向有其需要的患者施用治疗有效量的权利要求1-26的人工转录因子，其中待治疗疾病由特异性效应物与受体的结合所调节，为此所述多指锌指蛋白特异性靶向所述受体基因启动子。
28.治疗由内皮素调节的疾病的方法，其包括向有其需要的患者施用治疗有效量的权利要求2、3或6-17的人工转录因子。
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| EP11184706.7 | 2011-10-11 | ||
| EP11184706 | 2011-10-11 | ||
| PCT/EP2012/069981 WO2013053719A2 (en) | 2011-10-11 | 2012-10-10 | Regulation of receptor expression through delivery of artificial transcription factors |
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| EP (1) | EP2766484A2 (zh) |
| JP (1) | JP2014530607A (zh) |
| KR (1) | KR20140079780A (zh) |
| CN (1) | CN103998609A (zh) |
| AU (1) | AU2012323032A1 (zh) |
| BR (1) | BR112014008456A2 (zh) |
| CA (1) | CA2851560A1 (zh) |
| CL (1) | CL2014000897A1 (zh) |
| CO (1) | CO6930308A2 (zh) |
| EA (1) | EA201490531A1 (zh) |
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| IL (1) | IL231865A0 (zh) |
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| MA (1) | MA36970A1 (zh) |
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| KR20200086670A (ko) | 2017-10-13 | 2020-07-17 | 셀렉타 바이오사이언시즈, 인크. | 항바이러스 전달 벡터 IgM 반응을 약화시키기 위한 방법 및 조성물 |
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| MX2021014566A (es) | 2019-05-28 | 2022-03-22 | Selecta Biosciences Inc | Métodos y composiciones para la respuesta inmune atenuada anti-vector de transferencia viral. |
| WO2021169333A1 (zh) * | 2020-02-25 | 2021-09-02 | 四川省人民医院 | Znf124基因在视网膜色素变性疾病早期筛查或辅助诊断中的应用 |
| US11897888B1 (en) | 2020-04-30 | 2024-02-13 | Stinginn Llc | Small molecular inhibitors of sting signaling compositions and methods of use |
| WO2023064367A1 (en) | 2021-10-12 | 2023-04-20 | Selecta Biosciences, Inc. | Methods and compositions for attenuating anti-viral transfer vector igm responses |
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|---|---|---|---|---|
| CN111304314A (zh) * | 2020-02-25 | 2020-06-19 | 四川省人民医院 | Znf124基因在视网膜色素变性疾病早期筛查或辅助诊断中的应用 |
| CN111304314B (zh) * | 2020-02-25 | 2020-11-20 | 四川省人民医院 | Znf124基因在视网膜色素变性疾病早期筛查或辅助诊断中的应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| KR20140079780A (ko) | 2014-06-27 |
| EA201490531A1 (ru) | 2014-08-29 |
| AU2012323032A1 (en) | 2014-04-03 |
| IN2014CN02586A (zh) | 2015-08-07 |
| CL2014000897A1 (es) | 2014-11-21 |
| MX2014004331A (es) | 2014-11-26 |
| TN2014000117A1 (en) | 2015-07-01 |
| HK1197083A1 (zh) | 2015-01-02 |
| SG11201400701WA (en) | 2014-08-28 |
| MA36970A1 (fr) | 2016-03-31 |
| CO6930308A2 (es) | 2014-04-28 |
| EP2766484A2 (en) | 2014-08-20 |
| WO2013053719A2 (en) | 2013-04-18 |
| US20140296129A1 (en) | 2014-10-02 |
| IL231865A0 (en) | 2014-05-28 |
| JP2014530607A (ja) | 2014-11-20 |
| ZA201401960B (en) | 2015-06-24 |
| CA2851560A1 (en) | 2013-04-18 |
| BR112014008456A2 (pt) | 2017-04-11 |
| WO2013053719A3 (en) | 2013-06-27 |
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