CN112105728B - CRISPR/Cas效应蛋白及*** - Google Patents

CRISPR/Cas效应蛋白及*** Download PDF

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CN112105728B
CN112105728B CN201980030881.2A CN201980030881A CN112105728B CN 112105728 B CN112105728 B CN 112105728B CN 201980030881 A CN201980030881 A CN 201980030881A CN 112105728 B CN112105728 B CN 112105728B
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赖锦盛
周英思
朱金洁
张湘博
赵海铭
宋伟彬
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Abstract

涉及核酸编辑领域，特别是规律成簇的间隔短回文重复(CRISPR)技术领域。一种Cas效应蛋白，包含此类蛋白的融合蛋白，以及编码它们的核酸分子。提供了用于核酸编辑(例如，基因或基因组编辑)的复合物和组合物，其包含上述蛋白或融合蛋白，或编码它们的核酸分子。用于核酸编辑(例如，基因或基因组编辑)的方法，其使用包含上述的蛋白或融合蛋白。

Description

CRISPR/Cas效应蛋白及***
技术领域
本发明涉及核酸编辑领域，特别是规律成簇的间隔短回文重复(CRISPR)技术领域。具体而言，本发明涉及Cas效应蛋白，包含此类蛋白的融合蛋白，以及编码它们的核酸分子。本发明还涉及用于核酸编辑(例如，基因或基因组编辑)的复合物和组合物，其包含本发明的蛋白或融合蛋白，或编码它们的核酸分子。本发明还涉及用于核酸编辑(例如，基因或基因组编辑)的方法，其使用包含本发明的蛋白或融合蛋白。
背景技术
CRISPR/Cas技术是一种被广泛使用的基因编辑技术，它通过RNA引导对基因组上的靶序列进行特异性结合并切割DNA产生双链断裂，利用生物非同源末端连接或同源重组进行定点基因编辑。
CRISPR/Cas9***是最常用的II型CRISPR***，它识别3’-NGG的PAM基序，对靶标序列进行平末端切割。CRISPR/Cas Type V***是一类近两年新发现的的CRISPR***，它具有5’-TTN的基序，对靶标序列进行粘性末端切割，例如Cpf1,C2c1,CasX,CasY。然而目前存在的不同的CRISPR/Cas各有不同的优点和缺陷。例如Cas9,C2c1和CasX均需要两条RNA进行向导RNA，而Cpf1只需要一条向导RNA而且可以用来进行多重基因编辑。CasX具有980个氨基酸的大小，而常见的Cas9，C2c1,CasY和Cpf1通常大小在1300个氨基酸左右。此外，Cas9，Cpf1，CasX，CasY的PAM序列都比较复杂多样，而C2c1识别严谨的5’-TTN,因此它的靶标位点比其他***容易被预测从而降低了潜在的脱靶效应。
总之，鉴于目前可获得的CRISPR/Cas***都受限于一些缺陷，开发一种更稳健的、具有多方面良好性能的新型CRISPR/Cas***对生物技术的发展具有重要意义。
发明内容
本申请的发明人经过大量实验和反复摸索，出人意料地发现了一种新型RNA指导的核酸内切酶。基于这一发现，本发明人开发了新的CRISPR/Cas***以及基于该***的基因编辑方法。
Cas效应蛋白
因此，在第一方面，本发明提供了一种蛋白，其具有SEQ ID NOs：1-18任一项所示的氨基酸序列或其直系同源物、同源物、变体或功能性片段；其中，所述直系同源物、同源物、变体或功能性片段基本保留了其所源自的序列的生物学功能。
在某些实施方案中，所述蛋白具有Cas效应蛋白活性。在某些实施方案中，所述蛋白是CRISPR/Cas***中的效应蛋白。
在本发明中，上述序列的生物学功能是指Cas效应蛋白活性，包括但不限于，与导向RNA结合的活性、核酸内切酶活性、在导向RNA引导下与靶序列特定位点结合并切割的活性。
在某些实施方案中，所述直系同源物、同源物、变体与其所源自的序列相比具有至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、或至少99％的序列同一性，并且基本保留了其所源自的序列的Cas效应蛋白活性(例如，与导向RNA结合的活性、核酸内切酶活性、在导向RNA引导下与靶序列特定位点结合并切割的活性)。
在某些实施方案中，所述直系同源物、同源物、变体与SEQ ID NOs：1-18任一项所示的序列相比具有至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、或至少99％的序列同一性，并且基本保留了其所源自的序列的Cas效应蛋白活性(例如，与导向RNA结合的活性、核酸内切酶活性、在导向RNA引导下与靶序列特定位点结合并切割的活性)。
在某些实施方案中，所述蛋白来自选自下列的物种：Sulfuricurvum sp、Omnitrophica WOR_2、Smithella sp和Agrobacterium sp。在某些实施方案中，所述蛋白为包含于选自下列的物种的CRISPR基因座(CRISPR locus)中的Cas效应蛋白：Sulfuricurvumsp、Omnitrophica WOR_2、Smithella sp和Agrobacterium sp。在此类实施方案中，所述蛋白具有SEQ ID NOs：5-18任一项所示的氨基酸序列或其直系同源物、同源物、变体或功能性片段；其中，所述直系同源物、同源物、变体或功能性片段基本保留了其所源自的序列的生物学功能。在某些实施方案中，所述直系同源物、同源物、变体与SEQ ID NOs：5-18任一项所示的氨基酸序列相比具有至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、或至少99％的序列同一性，并且基本保留了其所源自的序列的Cas效应蛋白活性(例如，与导向RNA结合的活性、核酸内切酶活性、在导向RNA引导下与靶序列特定位点结合并切割的活性)。
在某些实施方案中，本发明的蛋白包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NOs：1-18任一项所示的序列；
(ii)与SEQ ID NOs：1-18任一项所示的序列相比具有一个或多个氨基酸的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个氨基酸的置换、缺失或添加)的序列；或
(iii)与SEQ ID NOs：1-18任一项所示的序列具有至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、或至少99％的序列同一性的序列。
在某些实施方案中，本发明的蛋白包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：1或2所示的序列；
(ii)与SEQ ID NO：1或2所示的序列相比具有一个或多个氨基酸的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个氨基酸的置换、缺失或添加)的序列；或
(iii)与SEQ ID NO：1或2所示的序列具有至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、或至少99％的序列同一性的序列。
在某些实施方案中，本发明的蛋白具有SEQ ID NO：1或2所示的氨基酸序列。
在某些实施方案中，本发明的蛋白包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：3或4所示的序列；
(ii)与SEQ ID NO：3或4所示的序列相比具有一个或多个氨基酸的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个氨基酸的置换、缺失或添加)的序列；或
(iii)与SEQ ID NO：3或4所示的序列具有至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、或至少99％的序列同一性的序列。
在某些实施方案中，本发明的蛋白具有SEQ ID NO：3或4所示的氨基酸序列。
在某些实施方案中，本发明的蛋白包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：5或6所示的序列；
(ii)与SEQ ID NO：5或6所示的序列相比具有一个或多个氨基酸的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个氨基酸的置换、缺失或添加)的序列；或
(iii)与SEQ ID NO：5或6所示的序列具有至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、或至少99％的序列同一性的序列。
在某些实施方案中，本发明的蛋白具有SEQ ID NO：5或6所示的氨基酸序列。
在某些实施方案中，本发明的蛋白包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NOs：7-18任一项所示的序列；
(ii)与SEQ ID NOs：7-18任一项所示的序列相比具有一个或多个氨基酸的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个氨基酸的置换、缺失或添加)的序列；或
(iii)与SEQ ID NOs：7-18任一项所示的序列具有至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、或至少99％的序列同一性的序列。
在某些实施方案中，本发明的蛋白具有SEQ ID NOs：7-18任一项所示的氨基酸序列。
衍生的蛋白
本发明的蛋白可进行衍生化，例如被连接至另一个分子(例如另一个多肽或蛋白)。通常，蛋白的衍生化(例如，标记)不会不利影响该蛋白的期望活性(例如，与导向RNA结合的活性、核酸内切酶活性、在导向RNA引导下与靶序列特定位点结合并切割的活性)。因此，本发明的蛋白还意欲包括此类衍生化的形式。例如，可以将本发明的蛋白功能性连接(通过化学偶合、基因融合、非共价连接或其它方式)于一个或多个其它分子基团，例如另一个蛋白或多肽，检测试剂，药用试剂等。
特别地，可以将本发明的蛋白连接其他功能性单元。例如，可以将其与核定位信号(NLS)序列连接，以提高本发明的蛋白进入细胞核的能力。例如，可以将其与靶向部分连接，以使得本发明的蛋白具有靶向性。例如，可以将其与可检测的标记连接，以便于对本发明的蛋白进行检测。例如，可以将其与表位标签连接，以便于本发明的蛋白的表达、检测、示踪和/或纯化。
缀合物
因此，在第二方面，本发明提供了一种缀合物，其包含如上所述的蛋白和修饰部分。
在某些实施方案中，所述修饰部分选自另外的蛋白或多肽、可检测的标记或其任意组合。
在某些实施方案中，所述另外的蛋白或多肽选自表位标签、报告基因序列、核定位信号(NLS)序列、靶向部分、转录激活结构域(例如，VP64)、转录抑制结构域(例如，KRAB结构域或SID结构域)、核酸酶结构域(例如，Fok1)，具有选自下列的活性的结构域：甲基化酶活性,去甲基化酶,转录激活活性,转录抑制活性,转录释放因子活性,组蛋白修饰活性,核酸酶活性,单链RNA切割活性,双链RNA切割活性,单链DNA切割活性,双链DNA切割活性和核酸结合活性；以及其任意组合。
在某些实施方案中，本发明的缀合物包含一个或多个NLS序列，例如SV40病毒大T抗原的NLS。在某些示例性实施方案中，所述NLS序列如SEQ ID NO:73所示。在某些实施方案中，所述NLS序列位于、靠近或接近本发明的蛋白的末端(例如，N端或C端)。在某些示例性实施方案中，所述NLS序列位于、靠近或接近本发明的蛋白的C端。
在某些实施方案中，本发明的缀合物包含表位标签(epitope tag)。这类表位标签是本领域技术人员熟知的，其实例包括但不限于His、V5、FLAG、HA、Myc、VSV-G、Trx等，并且本领域技术人员已知如何根据期望目的(例如，纯化、检测或示踪)选择合适的表位标签。
在某些实施方案中，本发明的缀合物包含报告基因序列。这类报告基因是本领域技术人员熟知的，其实例包括但不限于GST、HRP、CAT、GFP、HcRed、DsRed、CFP、YFP、BFP等。
在某些实施方案中，本发明的缀合物包含能够与DNA分子或细胞内分子结合的结构域，例如麦芽糖结合蛋白(MBP)、Lex A的DNA结合结构域(DBD)、GAL4的DBD等。
在某些实施方案中，本发明的缀合物包含可检测的标记，例如荧光染料，例如FITC或DAPI。
在某些实施方案中，本发明的蛋白任选地通过接头与所述修饰部分偶联、缀合或融合。
在某些实施方案中，所述修饰部分直接连接至本发明的蛋白的N端或C端。
在某些实施方案中，所述修饰部分通过接头连接至本发明的蛋白的N端或C端。这类接头是本领域熟知的，其实例包括但不限于包含一个或多个(例如，1个，2个，3个，4个或5个)氨基酸(如，Glu或Ser)或氨基酸衍生物(如，Ahx、β-Ala、GABA或Ava)的接头，或PEG等。
融合蛋白
在第三方面，本发明提供了一种融合蛋白，其包含本发明的蛋白以及另外的蛋白或多肽。
在某些实施方案中，所述另外的蛋白或多肽选自表位标签、报告基因序列、核定位信号(NLS)序列、靶向部分、转录激活结构域(例如，VP64)、转录抑制结构域(例如，KRAB结构域或SID结构域)、核酸酶结构域(例如，Fok1)，具有选自下列的活性的结构域：甲基化酶活性,去甲基化酶,转录激活活性,转录抑制活性,转录释放因子活性,组蛋白修饰活性,核酸酶活性,单链RNA切割活性,双链RNA切割活性,单链DNA切割活性,双链DNA切割活性和核酸结合活性；以及其任意组合。
在某些实施方案中，本发明的融合蛋白包含一个或多个NLS序列，例如SV40病毒大T抗原的NLS。在某些实施方案中，所述NLS序列位于、靠近或接近本发明的蛋白的末端(例如，N端或C端)。在某些示例性实施方案中，所述NLS序列位于、靠近或接近本发明的蛋白的C端。
在某些实施方案中，本发明的融合蛋白包含表位标签。
在某些实施方案中，本发明的融合蛋白包含报告基因序列。
在某些实施方案中，本发明的融合蛋白包含能够与DNA分子或细胞内分子结合的结构域。
在某些实施方案中，本发明的蛋白任选地通过接头与所述另外的蛋白或多肽融合。
在某些实施方案中，所述另外的蛋白或多肽直接连接至本发明的蛋白的N端或C端。
在某些实施方案中，所述另外的蛋白或多肽通过接头连接至本发明的蛋白的N端或C端。
在某些示例性实施方案中，本发明的融合蛋白具有选自下列的氨基酸序列：SEQID NOs:74-91。
本发明的蛋白、本发明的缀合物或本发明的融合蛋白不受其产生方式的限定，例如，其可以通过基因工程方法(重组技术)产生，也可以通过化学合成方法产生。
同向重复序列
在第四方面，本发明提供了一种分离的核酸分子，其包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NOs：37-54任一项所示的序列；
(ii)与SEQ ID NOs：37-54任一项所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NOs：37-54任一项所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列；
并且，(ii)-(v)中任一项所述的序列基本保留了其所源自的序列的生物学功能，所述序列的生物学功能是指，作为CRISPR-Cas***中的同向重复序列的活性。
在某些实施方案中，所述分离的核酸分子是CRISPR-Cas***中的同向重复序列。
在某些实施方案中，所述核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NOs：37-54任一项所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；或
(c)(a)中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子是RNA。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：37所示的序列；
(ii)与SEQ ID NO：37所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：37所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：37所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；或
(c)SEQ ID NO：37所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：38所示的序列；
(ii)与SEQ ID NO：38所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：38所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：38所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；或
(c)SEQ ID NO：38所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：39所示的序列；
(ii)与SEQ ID NO：39所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：39所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：39所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：39所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：40所示的序列；
(ii)与SEQ ID NO：40所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：40所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：40所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：40所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：41所示的序列；
(ii)与SEQ ID NO：41所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：41所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：41所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：41所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：42所示的序列；
(ii)与SEQ ID NO：42所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：42所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：42所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：42所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：43所示的序列；
(ii)与SEQ ID NO：43所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：43所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：43所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：43所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：44所示的序列；
(ii)与SEQ ID NO：44所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：44所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：44所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：44所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：45所示的序列；
(ii)与SEQ ID NO：45所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：45所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：45所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：45所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：46所示的序列；
(ii)与SEQ ID NO：46所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：46所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：46所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：46所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：47所示的序列；
(ii)与SEQ ID NO：47所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：47所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：47所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：47所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：48所示的序列；
(ii)与SEQ ID NO：48所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：48所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：48所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：48所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：49所示的序列；
(ii)与SEQ ID NO：49所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：49所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：49所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：49所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：50所示的序列；
(ii)与SEQ ID NO：50所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：50所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：50所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：50所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：51所示的序列；
(ii)与SEQ ID NO：51所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：51所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：51所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：51所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：52所示的序列；
(ii)与SEQ ID NO：52所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：52所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：52所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：52所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：53所示的序列；
(ii)与SEQ ID NO：53所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：53所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：53所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：53所示的核苷酸序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(i)SEQ ID NO：54所示的序列；
(ii)与SEQ ID NO：54所示的序列相比具有一个或多个碱基的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个碱基的置换、缺失或添加)的序列；
(iii)与SEQ ID NO：54所示的序列具有至少20％、至少30％、至少40％、至少50％、至少60％、至少70％、至少80％、至少90％、至少95％的序列同一性的序列；
(iv)在严格条件下与(i)-(iii)任一项中所述的序列杂交的序列；或
(v)(i)-(iii)任一项中所述的序列的互补序列。
在某些实施方案中，所述分离的核酸分子包含选自下列的序列，或由选自下列的序列组成：
(a)SEQ ID NO：54所示的核苷酸序列；
(b)在严格条件下与(a)中所述的序列杂交的序列；
(c)SEQ ID NO：54所示的核苷酸序列的互补序列。
CRISPR/Cas复合物
在第五方面，本发明提供了一种复合物，其包含：
(i)蛋白组分，其选自：本发明的蛋白、缀合物或融合蛋白，及其任意组合；和
(ii)核酸组分，其从5’至3’方向包含如第四方面所述的分离的核酸分子和能够与靶序列杂交的导向序列，
其中，所述蛋白组分与核酸组分相互结合形成复合物。
在某些实施方案中，所述导向序列连接于所述核酸分子的3’端。
在某些实施方案中，所述导向序列包含所述靶序列的互补序列。
在某些实施方案中，所述核酸组分是CRISPR-Cas***中的导向RNA。
在某些实施方案中，所述核酸分子是RNA。
在某些实施方案中，所述复合物不包含反式作用crRNA(tracrRNA)。
在某些实施方案中，所述导向序列在长度上为至少5个、至少10个、在某些实施方案中，所述导向序列在长度上为10-30个、或15-25个、或15-22个、或19-25个或19-22个核苷酸。
在某些实施方案中，所述分离的核酸分子在长度上为55-70个核苷酸，例如55-65个核苷酸，例如60-65个核苷酸，例如62-65个核苷酸，例如63-64个核苷酸。在某些实施方案中，所述分离的核酸分子在长度上为15-30个核苷酸，例如15-25个核苷酸，例如20-25个核苷酸，例如22-24个核苷酸，例如23个核苷酸。
编码核酸、载体及宿主细胞
在第六方面，本发明提供了一种分离的核酸分子，其包含：
(i)编码本发明的蛋白或融合蛋白的核苷酸序列；
(ii)编码如第四方面所述的分离的核酸分子；或
(iii)包含(i)和(ii)的核苷酸序列。
在某些实施方案中，(i)-(iii)任一项中所述的核苷酸序列经密码子优化用于在原核细胞中进行表达。在某些实施方案中，(i)-(iii)任一项中所述的核苷酸序列经密码子优化用于在真核细胞中进行表达。
在第七方面，本发明还提供了一种载体，其包含如第六方面所述的分离的核酸分子。本发明的载体可以是克隆载体，也可以是表达载体。在某些实施方案中，本发明的载体是例如质粒，粘粒，噬菌体，柯斯质粒等等。在某些选实施方案中，所述载体能够在受试者(例如哺乳动物，例如人)体内表达本发明的蛋白、融合蛋白、如第四方面所述的分离的核酸分子或如第五方面所述的复合物。
在第八方面，本发明还提供了包含如上所述的分离的核酸分子或载体的宿主细胞。此类宿主细胞包括但不限于，原核细胞例如大肠杆菌细胞，以及真核细胞例如酵母细胞，昆虫细胞，植物细胞和动物细胞(如哺乳动物细胞，例如小鼠细胞、人细胞等)。本发明的细胞还可以是细胞系，例如293T细胞。
组合物及载体组合物
在第九方面，本发明还提供了一种组合物，其包含：
(i)第一组分，其选自：本发明的蛋白、缀合物、融合蛋白、编码所述蛋白或融合蛋白的核苷酸序列，以及其任意组合；和
(ii)第二组分，其为包含导向RNA的核苷酸序列，或者编码所述包含导向RNA的核苷酸序列的核苷酸序列；
其中，所述导向RNA从5’至3’方向包含同向重复序列和导向序列，所述导向序列能够与靶序列杂交；
所述导向RNA能够与(i)中所述的蛋白、缀合物或融合蛋白形成复合物。
在某些实施方案中，所述同向重复序列是如第四方面所定义的分离的核酸分子。
在某些实施方案中，所述导向序列连接至所述同向重复序列的3’端。在某些实施方案中，所述导向序列包含所述靶序列的互补序列。
在某些实施方案中，所述组合物不包含tracrRNA。
在某些实施方案中，所述组合物是非天然存在的或经修饰的。在某些实施方案中，所述组合物中的至少一个组分是非天然存在的或经修饰的。在某些实施方案中，所述第一组分是非天然存在的或经修饰的；和/或，所述第二组分是非天然存在的或经修饰的。
在某些实施方案中，当所述靶序列为DNA时，所述靶序列位于原间隔序列临近基序(PAM)的3’端，并且所述PAM具有5’-NTN或5’-TNN所示的序列，其中，N选自A、G、T、C。
在某些实施方案中，当所述靶序列为RNA时，所述靶序列不具有PAM结构域限制。
在某些实施方案中，所述靶序列是来自原核细胞或真核细胞的DNA或RNA序列。在某些实施方案中，所述靶序列是非天然存在的DNA或RNA序列。
在某些实施方案中，所述靶序列存在于细胞内。在某些实施方案中，所述靶序列存在于细胞核内或细胞质(例如，细胞器)内。在某些实施方案中，所述细胞是真核细胞。在某些实施方案中，所述细胞是原核细胞。
在某些实施方案中，所述蛋白连接有一个或多个NLS序列。在某些实施方案中，所述缀合物或融合蛋白包含一个或多个NLS序列。在某些实施方案中，所述NLS序列连接至所述蛋白的N端或C端。在某些实施方案中，所述NLS序列融合至所述蛋白的N端或C端。
在第十方面，本发明还提供了一种组合物，其包含一种或多种载体，所述一种或多种载体包含：
(i)第一核酸，其为编码本发明的蛋白或融合蛋白的核苷酸序列；任选地所述第一核酸可操作地连接至第一调节元件；以及
(ii)第二核酸，其编码包含导向RNA的核苷酸序列；任选地所述第二核酸可操作地连接至第二调节元件；
其中：
所述第一核酸与第二核酸存在于相同或不同的载体上；
所述导向RNA从5’至3’方向包含同向重复序列和导向序列，所述导向序列能够与靶序列杂交；
所述导向RNA能够与(i)中所述的效应蛋白或融合蛋白形成复合物。
在某些实施方案中，所述同向重复序列是如第四方面所定义的分离的核酸分子。
在某些实施方案中，所述导向序列连接至所述同向重复序列的3’端。在某些实施方案中，所述导向序列包含所述靶序列的互补序列。
在某些实施方案中，所述组合物不包含tracrRNA。
在某些实施方案中，所述组合物是非天然存在的或经修饰的。在某些实施方案中，所述组合物中的至少一个组分是非天然存在的或经修饰的。
在某些实施方案中，所述第一调节元件是启动子，例如诱导型启动子。
在某些实施方案中，所述第二调节元件是启动子，例如诱导型启动子。
在某些实施方案中，当所述靶序列为DNA时，所述靶序列位于原间隔序列临近基序(PAM)的3’端，并且所述PAM具有5’-NTN或5’-TNN所示的序列，其中，N选自A、G、T、C。
在某些实施方案中，当所述靶序列为RNA时，所述靶序列不具有PAM结构域限制。
在某些实施方案中，所述靶序列是来自原核细胞或真核细胞的DNA或RNA序列。在某些实施方案中，所述靶序列是非天然存在的DNA或RNA序列。
在某些实施方案中，所述靶序列存在于细胞内。在某些实施方案中，所述靶序列存在于细胞核内或细胞质(例如，细胞器)内。在某些实施方案中，所述细胞是真核细胞。在某些实施方案中，所述细胞是原核细胞。
在某些实施方案中，所述蛋白连接有一个或多个NLS序列。在某些实施方案中，所述缀合物或融合蛋白包含一个或多个NLS序列。在某些实施方案中，所述NLS序列连接至所述蛋白的N端或C端。在某些实施方案中，所述NLS序列融合至所述蛋白的N端或C端。
在某些实施方案中，一种类型的载体是质粒，其是指其中可以例如通过标准分子克隆技术***另外的DNA片段的环状双链DNA环。另一种类型的载体是病毒载体，其中病毒衍生的DNA或RNA序列存在于用于包装病毒(例如，逆转录病毒、复制缺陷型逆转录病毒、腺病毒、复制缺陷型腺病毒、以及腺相关病毒)的载体中。病毒载体还包含由用于转染到一种宿主细胞中的病毒携带的多核苷酸。某些载体(例如，具有细菌复制起点的细菌载体和附加型哺乳动物载体)能够在它们被导入的宿主细胞中自主复制。其他载体(例如，非附加型哺乳动物载体)在引入宿主细胞后整合到该宿主细胞的基因组中，并且由此与该宿主基因组一起复制。而且，某些载体能够指导它们可操作连接的基因的表达。这样的载体在此被称为“表达载体”。在重组DNA技术中使用的普通表达栽体通常是质粒形式。
重组表达载体可包含处于适合于在宿主细胞中的核酸表达的形式的本发明的核酸分子，这意味着这些重组表达载体包含基于待用于表达的宿主细胞而选择的一种或多种调节元件，所述调节元件可操作地连接至待表达的核酸序列。
递送及递送组合物
本发明的蛋白、缀合物、融合蛋白、如第四方面所述的分离的核酸分子、本发明的复合物、如第六方面所述的分离的核酸分子、如第七方面所述的载体、如第九方面及第十方面所述的组合物，可以通过本领域已知的任何方法进行递送。此类方法包括但不限于，电穿孔、脂转染、核转染、显微注射、声孔效应、基因枪、磷酸钙介导的转染、阳离子转染、脂质体转染、树枝状转染、热激转染、核转染、磁转染、脂转染、穿刺转染、光学转染、试剂增强性核酸摄取、以及经由脂质体、免疫脂质体、病毒颗粒、人工病毒体等的递送。
因此，在另一个方面，本发明提供了一种递送组合物，其包含递送载体，以及选自下列的一种或多种：本发明的蛋白、缀合物、融合蛋白、如第四方面所述的分离的核酸分子、本发明的复合物、如第六方面所述的分离的核酸分子、如第七方面所述的载体、如第九方面及第十方面所述的组合物。
在某些实施方案中，所述递送载体是粒子。
在某些实施方案中，所述递送载体选自脂质颗粒、糖颗粒、金属颗粒、蛋白颗粒、脂质体、外泌体、微泡、基因枪或病毒载体(例如，复制缺陷型逆转录病毒、慢病毒、腺病毒或腺相关病毒)。
试剂盒
在另一个方面，本发明提供了一种试剂盒，其包含如上所述的组分中的一种或多种。在某些实施方案中，所述试剂盒包含一种或多种选自下列的组分：本发明的蛋白、缀合物、融合蛋白、如第四方面所述的分离的核酸分子、本发明的复合物、如第六方面所述的分离的核酸分子、如第七方面所述的载体、如第九方面及第十方面所述的组合物。
在某些实施方案中，本发明的试剂盒包含如第九方面所述的组合物。在某些实施方案中，所述试剂盒还包含使用所述组合物的说明书。
在某些实施方案中，本发明的试剂盒包含如第十方面所述的组合物。在某些实施方案中，所述试剂盒还包含使用所述组合物的说明书。
在某些实施方案中，本发明的试剂盒中包含的组分可以被提供于任何适合的容器中。
在某些实施方案中，所述试剂盒还包含一种或多种缓冲液。缓冲液可以是任何缓冲液，包括但不限于碳酸钠缓冲液、碳酸氢钠缓冲液、硼酸盐缓冲液、Tris缓冲液、MOPS缓冲液、HEPES缓冲液及其组合。在某些实施方案中，该缓冲液是碱性的。在某些实施方案中，该缓冲液具有从约7至约10的pH。
在某些实施方案中，该试剂盒还包括一个或多个寡核苷酸，该一个或多个寡核苷酸对应于一个用于***进载体中的导向序列，以便可操作地连接该导向序列和调节元件。在某些实施方案中，该试剂盒包括用于同源重组的编辑模板。
方法及用途
在另一个方面，本发明提供了一种修饰靶基因的方法，其包括：将如第五方面所述的复合物、如第九方面所述的组合物或如第十方面所述的组合物与所述靶基因接触，或者递送至包含所述靶基因的细胞中；所述靶序列存在于所述靶基因中。
在某些实施方案中，所述方法用于体外(in vitro)或离体(ex vivo)修饰靶基因。在某些实施方案中，所述方法不是通过疗法来治疗人或动物的方法。在某些实施方案中，所述方法不包括修饰人类种系遗传特性的步骤。
在某些实施方案中，所述靶基因存在于细胞内。在某些实施方案中，所述细胞是原核细胞。在某些实施方案中，所述细胞是真核细胞。在某些实施方案中，所述细胞是哺乳动物细胞。在某些实施方案中，所述细胞是人类细胞。在某些实施方案中，所述细胞选自非人灵长类动物、牛、猪或啮齿类动物细胞。在某些实施方案中，所述细胞是非哺乳动物真核细胞，例如家禽或鱼等。在某些实施方案中，所述细胞是植物细胞，例如栽培植物(如木薯、玉米、高粱、小麦或水稻)、藻类、树或蔬菜具有的细胞。
在某些实施方案中，所述靶基因存在于体外的核酸分子(例如，质粒)中。在某些实施方案中，所述靶基因存在于质粒中。
在某些实施方案中，所述修饰是指所述靶序列的断裂，如DNA的双链断裂或RNA的单链断裂。
在某些实施方案中，所述断裂导致靶基因的转录降低。
在某些实施方案中，所述方法还包括：将编辑模板与所述靶基因接触，或者递送至包含所述靶基因的细胞中。在此类实施方案中，所述方法通过与所述编辑模板同源重组修复所述断裂的靶基因，其中所述修复导致一种突变，包括所述靶基因的一个或多个核苷酸的***、缺失、或取代。在某些实施方案中，所述突变导致在从包含该靶序列的基因表达的蛋白质中的一个或多个氨基酸改变。
因此，在某些实施方案中，所述修饰还包括将编辑模板(例如外源核酸)***所述断裂中。
在某些实施方案中，所述的蛋白、缀合物、融合蛋白、分离的核酸分子、复合物、载体或组合物包含于递送载体中。
在某些实施方案中，所述递送载体选自脂质颗粒、糖颗粒、金属颗粒、蛋白颗粒、脂质体、外泌体、病毒载体(如复制缺陷型逆转录病毒、慢病毒、腺病毒或腺相关病毒)。
在某些实施方案中，所述方法其用于改变靶基因或编码靶基因产物的核酸分子中的一个或多个靶序列来修饰细胞、细胞系或生物体。
在另一个方面，本发明提供了一种改变基因产物的表达的方法，其包括：将如第五方面所述的复合物、如第九方面所述的组合物或如第十方面所述的组合物与编码所述基因产物的核酸分子接触，或者递送至包含所述核酸分子的细胞中，所述靶序列存在于所述核酸分子中。
在某些实施方案中，所述核酸分子存在于细胞内。在某些实施方案中，所述细胞是原核细胞。在某些实施方案中，所述细胞是真核细胞。在某些实施方案中，所述细胞是哺乳动物细胞。在某些实施方案中，所述细胞是人类细胞。在某些实施方案中，所述细胞选自非人灵长类动物、牛、猪或啮齿类动物细胞。在某些实施方案中，所述细胞是非哺乳动物真核细胞，例如家禽或鱼等。在某些实施方案中，所述细胞是植物细胞，例如栽培植物(如木薯、玉米、高粱、小麦或水稻)、藻类、树或蔬菜具有的细胞。
在某些实施方案中，所述核酸分子存在于体外的核酸分子(例如，质粒)中。在某些实施方案中，所述核酸分子存在于质粒中。
在某些实施方案中，所述基因产物的表达被改变(例如，增强或降低)。在某些实施方案中，所述基因产物的表达被增强。在某些实施方案中，所述基因产物的表达被降低。
在某些实施方案中，所述基因产物是蛋白。
在某些实施方案中，所述的蛋白、缀合物、融合蛋白、分离的核酸分子、复合物、载体或组合物包含于递送载体中。
在某些实施方案中，所述递送载体选自脂质颗粒、糖颗粒、金属颗粒、蛋白颗粒、脂质体、外泌体、病毒载体(如复制缺陷型逆转录病毒、慢病毒、腺病毒或腺相关病毒)。
在某些实施方案中，所述方法其用于改变靶基因或编码靶基因产物的核酸分子中的一个或多个靶序列来修饰细胞、细胞系或生物体。
在另一个方面，本发明涉及如第一方面所述的蛋白、如第二方面所述的缀合物、如第三方面所述的融合蛋白、如第四方面所述的分离的核酸分子、如第五方面所述的复合物、如第六方面所述的分离的核酸分子、如第七方面所述的载体、如第九方面所述的组合物、如第十方面所述的组合物、本发明的试剂盒或递送组合物，用于核酸编辑(例如，体外或离体核酸编辑)的用途，或者在制备制剂中的用途，所述制剂用于核酸编辑。
在某些实施方案中，待被编辑的核酸存在于细胞内。在某些实施方案中，所述细胞是原核细胞或真核细胞。在某些实施方案中，待被编辑的核酸存在于体外的核酸分子(例如，质粒)中。
在某些实施方案中，所述核酸编辑包括基因或基因组编辑，例如修饰基因、敲除基因、改变基因产物的表达、修复突变、和/或***多核苷酸。在某些实施方案中，所述基因或基因组编辑不包括修饰人类种系遗传特性的步骤。在某些实施方案中，所述用途不是通过疗法来治疗人或动物的方法。
在某些实施方案中，所述用途还包括通过与外源模板多核苷酸的同源重组来修复被编辑的靶序列，其中所述修复可以产生该靶序列的突变，包括一个或多个核苷酸的***、缺失或取代。
在另一个方面，本发明涉及如第一方面所述的蛋白、如第二方面所述的缀合物、如第三方面所述的融合蛋白、如第四方面所述的分离的核酸分子、如第五方面所述的复合物、如第六方面所述的分离的核酸分子、如第七方面所述的载体、如第九方面所述的组合物、如第十方面所述的组合物、本发明的试剂盒或递送组合物，在制备制剂中的用途，所述制剂用于：
(i)体外或离体单链DNA的检测(例如原核细胞中的单链DNA的检测)；
(ii)编辑靶基因座中的靶序列来修饰生物或非人类生物(例如，原核生物)。
细胞及细胞子代
在某些情况下，由本发明的方法引入到细胞的修饰可以使得细胞和其子代被改变以改进其生物产物(如抗体、淀粉、乙醇或其他期望的细胞输出物)的产生。在某些情况下，由本发明的方法引入到细胞的修饰可以使得细胞和其子代包括使所生产生物产物发生变化的改变。
因此，在另一方面，本发明还涉及如上所述的方法获得的细胞或其子代，其中所述细胞含有在其野生型中不存在的修饰。
本发明还涉及如上所述的细胞或其子代的细胞产物。
本发明还涉及一种体外的、离体的或体内的细胞或细胞系或它们的子代，所述细胞或细胞系或它们的子代包含：如第一方面所述的蛋白、如第二方面所述的缀合物、如第三方面所述的融合蛋白、如第四方面所述的分离的核酸分子、如第五方面所述的复合物、如第六方面所述的分离的核酸分子、如第七方面所述的载体、如第九方面所述的组合物、如第十方面所述的组合物、本发明的试剂盒或递送组合物。
在某些实施方案中，所述细胞是原核细胞。
在某些实施方案中，所述细胞是真核细胞。在某些实施方案中，所述细胞是哺乳动物细胞。在某些实施方案中，所述细胞是人类细胞。某些实施方案中，所述细胞是非人哺乳动物细胞，例如非人灵长类动物、牛、羊、猪、犬、猴、兔、啮齿类(如大鼠或小鼠)的细胞。在某些实施方案中，所述细胞是非哺乳动物真核细胞，例如家禽鸟类(如鸡)、鱼类或甲壳动物(如蛤蜊、虾)的细胞。在某些实施方案中，所述细胞是植物细胞，例如单子叶植物或双子叶植物具有的细胞或栽培植物或粮食作物如木薯、玉米、高粱、大豆、小麦、燕麦或水稻具有的细胞，例如藻类、树或生产植物、果实或蔬菜(例如，树类如柑橘树、坚果树；茄属植物、棉花、烟草、番茄、葡萄、咖啡、可可等)。
在某些实施方案中，所述细胞是干细胞或干细胞系。
术语定义
在本发明中，除非另有说明，否则本文中使用的科学和技术名词具有本领域技术人员所通常理解的含义。并且，本文中所用的分子遗传学、核酸化学、化学、分子生物学、生物化学、细胞培养、微生物学、细胞生物学、基因组学和重组DNA等操作步骤均为相应领域内广泛使用的常规步骤。同时，为了更好地理解本发明，下面提供相关术语的定义和解释。
在本发明中，表述“Cas12g”是指，本发明人首次发现并鉴定的一种Cas效应蛋白，其具有选自下列的氨基酸序列：
(i)SEQ ID NO：1或2所示的序列；
(ii)与SEQ ID NO：1或2所示的序列相比具有一个或多个氨基酸的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个氨基酸的置换、缺失或添加)的序列；或
(iii)与SEQ ID NO：1或2所示的序列具有至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、或至少99％的序列同一性的序列。
本发明的Cas12g是一种在导向RNA引导下与靶序列特定位点结合并切割的核酸内切酶，同时具有DNA和RNA内切酶活性。
在本发明中，表述“Cas12h”是指，本发明人首次发现并鉴定的一种Cas效应蛋白，其具有选自下列的氨基酸序列：
(i)SEQ ID NO：3或4所示的序列；
(ii)与SEQ ID NO：3或4所示的序列相比具有一个或多个氨基酸的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个氨基酸的置换、缺失或添加)的序列；或
(iii)与SEQ ID NO：3或4所示的序列具有至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、或至少99％的序列同一性的序列。
本发明的Cas12h是一种在导向RNA引导下与靶序列特定位点结合并切割的核酸内切酶，同时具有DNA和RNA内切酶活性。
在本发明中，表述“Cas12w”是指，本发明人首次发现并鉴定的一种Cas效应蛋白，其具有选自下列的氨基酸序列：
(i)SEQ ID NO：5或6所示的序列；
(ii)与SEQ ID NO：5或6所示的序列相比具有一个或多个氨基酸的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个氨基酸的置换、缺失或添加)的序列；或
(iii)与SEQ ID NO：5或6所示的序列具有至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、或至少99％的序列同一性的序列。
本发明的Cas12w是一种在导向RNA引导下与靶序列特定位点结合并切割的核酸内切酶，同时具有DNA和RNA内切酶活性。
在本发明中，表述“Cas12j”是指，本发明人首次发现并鉴定的一种Cas效应蛋白，其具有选自下列的氨基酸序列：
(i)SEQ ID NOs：7-18任一项所示的序列；
(ii)与SEQ ID NOs：7-18任一项所示的序列相比具有一个或多个氨基酸的置换、缺失或添加(例如1个，2个，3个，4个，5个，6个，7个，8个，9个或10个氨基酸的置换、缺失或添加)的序列；或
(iii)与SEQ ID NOs：7-18任一项所示的序列具有至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、或至少99％的序列同一性的序列。
本发明的Cas12j是一种在导向RNA引导下与靶序列特定位点结合并切割的核酸内切酶，同时具有DNA和RNA内切酶活性。
如本文中所使用的，术语“规律成簇的间隔短回文重复(CRISPR)-CRISPR-相关(Cas)(CRISPR-Cas)***”或“CRISPR***”可互换地使用并且具有本领域技术人员通常理解的含义，其通常包含与CRISPR相关(“Cas”)基因的表达有关的转录产物或其他元件，或者能够指导所述Cas基因活性的转录产物或其他元件。此类转录产物或其他元件可以包含编码Cas效应蛋白的序列和包含CRISPR RNA(crRNA)的导向RNA，以及在CRISPR-Cas9***中所含有的反式作用crRNA(tracrRNA)序列，或来自CRISPR基因座的其他序列或转录产物。
如本文中所使用的，术语“Cas效应蛋白”、“Cas效应酶”可互换地使用并且是指，CRISPR-Cas***中呈现的任一种大于长度900个氨基酸的蛋白质。在某些情况下，这类蛋白是指从Cas基因座中鉴定的蛋白。
如本文中所使用的，术语“导向RNA(guide RNA)”、“成熟crRNA”可互换地使用并且具有本领域技术人员通常理解的含义。一般而言，导向RNA可以包含同向(direct)重复序列和导向序列(guide sequence)，或者基本上由或由同向重复序列和导向序列(在内源性CRISPR***背景下也称为间隔序列(spacer))组成。在某些情况下，导向序列是与靶序列具有足够互补性从而与所述靶序列杂交并引导CRISPR/Cas复合物与所述靶序列的特异性结合的任何多核苷酸序列。在某些实施方案中，当最佳比对时，导向序列与其相应靶序列之间的互补程度为至少50％、至少60％、至少70％、至少80％、至少90％、至少95％、或至少99％。确定最佳比对在本领域的普通技术人员的能力范围内。例如，存在公开和可商购的比对算法和程序，诸如但不限于ClustalW、matlab中的史密斯-沃特曼算法(Smith-Waterman)、Bowtie、Geneious、Biopython以及SeqMan。
在某些情况下，所述导向序列在长度上为至少5个、至少10个、至少15个、至少16个、至少17个、至少18个、至少19个、至少20个、至少21个、至少22个、至少23个、至少24个、至少25个、至少26个、至少27个、至少28个、至少29个、至少30个、至少35个、至少40个、至少45个或至少50个核苷酸。在某些情况下，所述导向序列在长度上为不超过50个、45个、40个、35个、30个、25个、24个、23个、22个、21个、20个、15个、10个或更少个核苷酸。在某些实施方案中，所述导向序列在长度上为10-30个、或15-25个、或15-22个、或19-25个或19-22个核苷酸。
在某些情况下，所述同向重复序列在长度上为至少10个、至少15个、至少16个、至少17个、至少18个、至少19个、至少20个、至少21个、至少22个、至少23个、至少24个、至少25个、至少26个、至少27个、至少28个、至少29个、至少30个、至少35个、至少40个、至少45个、至少50个、至少55个、至少56个、至少57个、至少58个、至少59个、至少60个、至少61个、至少62个、至少63个、至少64个、至少65个或至少70个核苷酸。在某些情况下，所述同向重复序列在长度上为不超过70个、65个、64个、63个、62个、61个、60个、59个、58个、57个、56个、55个、50个、45个、40个、35个、30个、29个、28个、27个、26个、25个、24个、23个、22个、21个、20个、15个、10个或更少个核苷酸。在某些实施方案中，所述同向重复序列在长度上为55-70个核苷酸，例如55-65个核苷酸，例如60-65个核苷酸，例如62-65个核苷酸，例如63-64个核苷酸。在某些实施方案中，所述同向重复序列在长度上为15-30个核苷酸，例如15-25个核苷酸，例如20-25个核苷酸，例如22-24个核苷酸，例如23个核苷酸。
如本文中所使用的，术语“CRISPR/Cas复合物”是指，导向RNA(guide RNA)或成熟crRNA与Cas蛋白结合所形成的核糖核蛋白复合体，其包含杂交到靶序列上并且与Cas蛋白结合的导向序列。该核糖核蛋白复合体能够识别并切割能与该导向RNA或成熟crRNA杂交的多核苷酸。
因此，在形成CRISPR/Cas复合物的情况下，“靶序列”是指被设计为具有靶向性的导向序列所靶向的多核苷酸，例如与该导向序列具有互补性的序列，其中靶序列与导向序列之间的杂交将促进CRISPR/Cas复合物的形成。完全互补性不是必需的，只要存在足够互补性以引起杂交并且促进一种CRISPR/Cas复合物的形成即可。靶序列可以包含任何多核苷酸，如DNA或RNA。在某些情况下，所述靶序列位于细胞的细胞核或细胞质中。在某些情况下，该靶序列可位于真核细胞的一个细胞器例如线粒体或叶绿体内。可被用于重组到包含该靶序列的靶基因座中的序列或模板被称为“编辑模板”或“编辑多核苷酸”或“编辑序列”。在某些实施方案中，所述编辑模板为外源核酸。在某些实施方案中，该重组是同源重组。
在本发明中，表述“靶序列”或“靶多核苷酸”可以是对细胞(例如，真核细胞)而言任何内源或外源的多核苷酸。例如，该靶多核苷酸可以是一种存在于真核细胞的细胞核中的多核苷酸。该靶多核苷酸可以是一个编码基因产物(例如，蛋白质)的序列或一个非编码序列(例如，调节多核苷酸或无用DNA)。在某些情况下，据信该靶序列应该与原间隔序列临近基序(PAM)相关。对PAM的精确序列和长度要求取决于使用的Cas效应酶而不同，但是PAM典型地是临近原间隔序列(也即，靶序列)的2-5个碱基对序列。本领域技术人员能够鉴定与给定的Cas效应蛋白一起使用的PAM序列。
在某些情况下，靶序列或靶多核苷酸可以包括多个疾病相关基因和多核苷酸以及信号传导生化途径相关基因和多核苷酸。此类靶序列或靶多核苷酸的非限制性实例，包括分别提交于2012年12月12日和2013年1月2日的美国临时专利申请61/736,527和61/748,427、提交于2013年12月12日的国际申请PCT/US2013/074667中所列举的那些，其全部通过引用并入本文。
在某些情况下，靶序列或靶多核苷酸的实例包括与信号传导生化途径相关的序列，例如信号传导生化途径相关基因或多核苷酸。靶多核苷酸的实例包括疾病相关基因或多核苷酸。“疾病相关”基因或多核苷酸是指与非疾病对照的组织或细胞相比，在来源于疾病影响的组织的细胞中以异常水平或以异常形式产生转录或翻译产物的任何基因或多核苷酸。在改变的表达与疾病的出现和/或进展相关的情况下，它可以是一个以异常高的水平被表达的基因；或者，它可以是一个以异常低的水平被表达的基因。疾病相关基因还指具有一个或多个突变或直接负责或与一个或多个负责疾病的病因学的基因连锁不平衡的遗传变异的基因。转录的或翻译的产物可以是已知的或未知的，并且可以处于正常或异常水平。
如本文中所使用的，术语“野生型”具有本领域技术人员通常理解的含义，其表示生物、菌株、基因的典型形式或者当它在自然界存在时区别于突变体或变体形式的特征，其可从自然中的来源分离并且没有被人为有意地修饰。
如本文中所使用的，术语“非天然存在的”或“工程化的”可互换地使用并且表示人工的参与。当这些术语用于描述核酸分子或多肽时，其表示该核酸分子或多肽至少基本上从它们在自然界中或如发现于自然界中的与其结合的至少另一种组分游离出来。
如本文中所使用的，术语“直系同源物(orthologue,ortholog)”具有本领域技术人员通常理解的含义。作为进一步指导，如本文中所述的蛋白质的“直系同源物”是指属于不同物种的蛋白质，该蛋白质执行与作为其直系同源物的蛋白相同或相似的功能。
如本文中所使用的，术语“同一性”用于指两个多肽之间或两个核酸之间序列的匹配情况。当两个进行比较的序列中的某个位置都被相同的碱基或氨基酸单体亚单元占据时(例如，两个DNA分子的每一个中的某个位置都被腺嘌呤占据，或两个多肽的每一个中的某个位置都被赖氨酸占据)，那么各分子在该位置上是同一的。两个序列之间的“百分数同一性”是由这两个序列共有的匹配位置数目除以进行比较的位置数目×100的函数。例如，如果两个序列的10个位置中有6个匹配，那么这两个序列具有60％的同一性。例如，DNA序列CTGACT和CAGGTT共有50％的同一性(总共6个位置中有3个位置匹配)。通常，在将两个序列比对以产生最大同一性时进行比较。这样的比对可通过使用，例如，可通过计算机程序例如Align程序(DNAstar,Inc.)方便地进行的Needleman等人(1970)J.Mol.Biol.48：443-453的方法来实现。还可使用已整合入ALIGN程序(版本2.0)的E.Meyers和W.Miller(Comput.ApplBiosci.，4:11-17(1988))的算法，使用PAM120权重残基表(weight residue table)、12的缺口长度罚分和4的缺口罚分来测定两个氨基酸序列之间的百分数同一性。此外，可使用已整合入GCG软件包(可在www.gcg.com上获得)的GAP程序中的Needleman和Wunsch(J MoIBiol.48:444-453(1970))算法，使用Blossum 62矩阵或PAM250矩阵以及16、14、12、10、8、6或4的缺口权重(gap weight)和1、2、3、4、5或6的长度权重来测定两个氨基酸序列之间的百分数同一性。
如本文中所使用的，术语“载体”是指，可将多聚核苷酸***其中的一种核酸运载工具。当载体能使***的多核苷酸编码的蛋白获得表达时，载体称为表达载体。载体可以通过转化，转导或者转染导入宿主细胞，使其携带的遗传物质元件在宿主细胞中获得表达。载体是本领域技术人员公知的，包括但不限于：质粒；噬菌粒；柯斯质粒；人工染色体，例如酵母人工染色体(YAC)、细菌人工染色体(BAC)或P1来源的人工染色体(PAC)；噬菌体如λ噬菌体或M13噬菌体及动物病毒等。可用作载体的动物病毒包括但不限于，逆转录酶病毒(包括慢病毒)、腺病毒、腺相关病毒、疱疹病毒(如单纯疱疹病毒)、痘病毒、杆状病毒、***瘤病毒、***多瘤空泡病毒(如SV40)。一种载体可以含有多种控制表达的元件，包括但不限于，启动子序列、转录起始序列、增强子序列、选择元件及报告基因。另外，载体还可含有复制起始位点。
如本文中所使用的，术语“宿主细胞”是指，可用于导入载体的细胞，其包括但不限于，如大肠杆菌或枯草菌等的原核细胞，如酵母细胞或曲霉菌等的真菌细胞，如S2果蝇细胞或Sf9等的昆虫细胞，或者如纤维原细胞，CHO细胞，COS细胞，NSO细胞，HeLa细胞，BHK细胞，HEK293细胞或人细胞等的动物细胞。
本领域技术人员将理解，表达载体的设计可取决于诸如待转化的宿主细胞的选择、所希望的表达水平等因素。一种载体可以被引入到宿主细胞中而由此产生转录物、蛋白质、或肽，包括由如本文所述的蛋白、融合蛋白、分离的核酸分子等(例如，CRISPR转录物，如核酸转录物、蛋白质、或酶)。
如本文中所使用的，术语“调节元件”旨在包括启动子、增强子、内部核糖体进入位点(IRES)、和其他表达控制元件(例如转录终止信号，如多聚腺苷酸化信号和多聚U序列)，其详细描述可参考戈德尔(Goeddel)，《基因表达技术：酶学方法》(GENE EXPRESSIONTECHNOLOGY:METHODS IN ENZYMOLOGY)185，学术出版社(Academic Press)，圣地亚哥(SanDiego)，加利福尼亚州(1990)。在某些情况下，调节元件包括指导一个核苷酸序列在许多类型的宿主细胞中的组成型表达的那些序列以及指导该核苷酸序列只在某些宿主细胞中表达的那些序列(例如，组织特异型调节序列)。组织特异型启动子可主要指导在感兴趣的期望组织中的表达，所述组织例如肌肉、神经元、骨、皮肤、血液、特定的器官(例如肝脏、胰腺)、或特殊的细胞类型(例如淋巴细胞)。在某些情况下，调节元件还可以时序依赖性方式(如以细胞周期依赖性或发育阶段依赖性方式)指导表达，该方式可以是或者可以不是组织或细胞类型特异性的。在某些情况下，术语“调节元件”涵盖的是增强子元件，如WPRE；CMV增强子；在HTLV-I的LTR中的R-U5’片段((Mol.Cell.Biol.，第8(1)卷，第466-472页，1988)；SV40增强子；以及在兔β-珠蛋白的外显子2与3之间的内含子序列(Proc.Natl.Acad.Sci.USA.，第78(3)卷，第1527-31页，1981)。
如本文中所使用的，术语“启动子”具有本领域技术人员公知的含义，其是指一段位于基因的上游能启动下游基因表达的非编码核苷酸序列。组成型(constitutive)启动子是这样的核苷酸序列：当其与编码或者限定基因产物的多核苷酸可操作地相连时，在细胞的大多数或者所有生理条件下，其导致细胞中基因产物的产生。诱导型启动子是这样的核苷酸序列，当可操作地与编码或者限定基因产物的多核苷酸相连时，基本上只有当对应于所述启动子的诱导物在细胞中存在时，其导致所述基因产物在细胞内产生。组织特异性启动子是这样的核苷酸序列：当可操作地与编码或者限定基因产物的多核苷酸相连时，基本上只有当细胞是该启动子对应的组织类型的细胞时，其才导致在细胞中产生基因产物。
如本文中所使用的，术语“可操作地连接”旨在表示感兴趣的核苷酸序列以一种允许该核苷酸序列的表达的方式被连接至该一种或多种调节元件(例如，处于一种体外转录/翻译***中或当该载体被引入到宿主细胞中时，处于该宿主细胞中)。
如本文中所使用的，术语“互补性”是指核酸与另一个核酸序列借助于传统的沃森-克里克或其他非传统类型形成一个或多个氢键的能力。互补百分比表示一个核酸分子中可与一个第二核酸序列形成氢键(例如，沃森-克里克碱基配对)的残基的百分比(例如，10个之中有5、6、7、8、9、10个即为50％、60％、70％、80％、90％、和100％互补)。“完全互补”表示一个核酸序列的所有连续残基与一个第二核酸序列中的相同数目的连续残基形成氢键。如本文使用的“基本上互补”是指在一个具有8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、30、35、40、45、50个或更多个核苷酸的区域上至少为60％、65％、70％、75％、80％、85％、90％、95％、97％、98％、99％、或100％的互补程度，或者是指在严格条件下杂交的两个核酸。
如本文中所使用的，对于杂交的“严格条件”是指与靶序列具有互补性的一个核酸主要地与该靶序列杂交并且基本上不杂交到非靶序列上的条件。严格条件通常是序列依赖性的，并且取决于许多因素而变化。一般而言，该序列越长，则该序列特异性地杂交到其靶序列上的温度就越高。严格条件的非限制性实例描述于蒂森(Tijssen)(1993)的《生物化学和分子生物学中的实验室技术-核酸探针杂交》(Laboratory Techniques InBiochemistryAnd Molecular Biology-Hybridization With Nucleic Acid Probes)，第I部分，第二章，“杂交原理概述和核酸探针分析策略”(“Overview of principles ofhybridization andthe strategy of nucleic acid probe assay”)，爱思唯尔(Elsevier)，纽约。
如本文中所使用的，术语“杂交”是指其中一个或多个多核苷酸反应形成一种复合物的反应，该复合物经由这些核苷酸残基之间的碱基的氢键键合而稳定化。氢键键合可以借助于沃森-克里克碱基配对、Hoogstein结合或以任何其他序列特异性方式而发生。该复合物可包含形成一个双链体的两条链、形成多链复合物的三条或多条链、单个自我杂交链、或这些的任何组合。杂交反应可以构成一个更广泛的过程(如PCR的开始、或经由一种酶的多核苷酸的切割)中的一个步骤。能够与一个给定序列杂交的序列被称为该给定序列的“互补物”。
如本文中所使用的，术语“表达”是指，藉此从DNA模板转录成多核苷酸(如转录成mRNA或其他RNA转录物)的过程和/或转录的mRNA随后藉此翻译成肽、多肽或蛋白质的过程。转录物和编码的多肽可以总称为“基因产物”。如果多核苷酸来源于基因组DNA，表达可以包括真核细胞中mRNA的剪接。
如本文中所使用的，术语“接头”是指，由多个氨基酸残基通过肽键连接形成的线性多肽。本发明的接头可以为人工合成的氨基酸序列，或天然存在的多肽序列，例如具有铰链区功能的多肽。此类接头多肽是本领域众所周知的(参见例如，Holliger,P.等人(1993)Proc.Natl.Acad.Sci.USA 90:6444-6448；Poljak,R.J.等人(1994)Structure 2:1121-1123)。
如本文中所使用的，术语“治疗”是指，治疗或治愈病症，延缓病症的症状的发作，和/或延缓病症的发展。
如本文中所使用的，术语“受试者”包括但不限于各种动物，例如哺乳动物，例如牛科动物、马科动物、羊科动物、猪科动物、犬科动物、猫科动物、兔科动物、啮齿类动物(例如，小鼠或大鼠)、非人灵长类动物(例如，猕猴或食蟹猴)或人。在某些实施方式中，所述受试者(例如人)患有病症(例如，疾病相关基因缺陷所导致的病症)。
发明的有益效果
与现有技术相比，本发明的Cas蛋白及***具有显著的有利方面。例如，本发明的Cas效应蛋白的PAM结构域具有的5’-TG结构，可以与其他PAM为5’-TTN的***相互补充，扩大识别范围。例如，本发明的Cas效应蛋白能在真核生物体内进行DNA切割，在分子大小上比Cpf1和Cas9蛋白小约200-400个氨基酸，因此转染效率上明显优于Cpf1和Cas9。
附图说明
图1为实施例3中Cas12h.1的PAM结构域分析结果。
序列信息
本发明涉及的部分序列的信息提供于下面的表1中。
表1：序列的描述
Figure BDA0002764893630000381
Figure BDA0002764893630000391
Figure BDA0002764893630000401
Figure BDA0002764893630000411
具体实施方式
现参照下列意在举例说明本发明(而非限定本发明)的实施例来描述本发明。
除非特别指明，否则基本上按照本领域内熟知的以及在各种参考文献中描述的常规方法进行实施例中描述的实验和方法。例如，本发明中所使用的免疫学、生物化学、化学、分子生物学、微生物学、细胞生物学、基因组学和重组DNA等常规技术，可参见参见萨姆布鲁克(Sambrook)、弗里奇(Fritsch)和马尼亚蒂斯(Maniatis)，《分子克隆：实验室手册》(MOLECULAR CLONING:A LABORATORY MANUAL)，第2次编辑(1989)；《当代分子生物学实验手册》(CURRENT PROTOCOLS IN MOLECULAR BIOLOGY)(F.M.奥苏贝尔(F.M.Ausubel)等人编辑，(1987))；《酶学方法》(METHODS IN ENZYMOLOGY)系列(学术出版公司)：《PCR 2：实用方法》(PCR 2:A PRACTICAL APPROACH)(M.J.麦克弗森(M.J.MacPherson)、B.D.黑姆斯(B.D.Hames)和G.R.泰勒(G.R.Taylor)编辑(1995))、哈洛(Harlow)和拉内(Lane)编辑(1988)《抗体：实验室手册》(ANTIBODIES,A LABORATORY MANUAL)，以及《动物细胞培养》(ANIMAL CELL CULTURE)(R.I.弗雷谢尼(R.I.Freshney)编辑(1987))。
另外，实施例中未注明具体条件者，按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者，均为可以通过市购获得的常规产品。本领域技术人员知晓，实施例以举例方式描述本发明，且不意欲限制本发明所要求保护的范围。本文中提及的全部公开案和其他参考资料以其全文通过引用合并入本文。
以下实施例涉及的部分试剂的来源如下：
LB液体培养基：10g胰蛋白胨(Tryptone)，5g酵母提取物(Yeast Extract)，10gNaCl，定容至1L，灭菌。若需加抗生素，则待培养基冷却后加，50μg/ml的终浓度。
氯仿/异戊醇：240ml的氯仿加10ml的异戊醇，混匀。
RNP缓冲液：100mM氯化钠,50mM Tris-HCl,10mM MgCl2,100μg/ml BSA,pH 7.9。
原核表达载体pACYC-Duet-1和pUC19购自北京全式金生物技术有限公司。
大肠杆菌感受态EC100购自Epicentre公司。
除非特别指明，以下实施例中涉及的序列合成均由南京金斯瑞生物科技有限公司完成，涉及的测序均由上海英骏生物技术有限公司完成。
实施例1.Cas12g基因和Cas12g导向RNA的获得
1、CRISPR和基因的注释：使用Prodigal对将NCBI和JGI数据库的微生物基因组和宏基因组数据进行基因注释得到所有蛋白，同时用Piler-CR进行CRISPR座的注释，参数均为默认参数。
2、蛋白质的过滤：通过序列一致性对注释蛋白去冗余，去除序列完全一致的蛋白，同时将长度大于800个氨基酸的蛋白划分为大分子蛋白。由于目前发现的所有第二类CRISPR/Cas***的效应蛋白长度多大于900个氨基酸，所以为了降低计算复杂度，我们在挖掘CRISPR效应蛋白的时候只对大分子蛋白进行考虑。
3、CRISPR相关大分子蛋白的获得：将每一个CRISPR座上下游延伸10Kb，将对CRISPR邻近区间内的非冗余大分子蛋白进行鉴定。
4、CRISPR相关大分子蛋白质的聚类：使用BLASTP对非冗余大分子CRISPR相关蛋白进行内部的两两比对，输出Evalue<1E-10的比对结果。使用MCL对BLASTP的输出结果进行聚类分析，CRISPR相关蛋白质家族。
5、CRISPR富集大分子蛋白质家族的鉴定：使用BLASTP对CRISPR相关蛋白质家族的蛋白比对到去除去CRISPR相关蛋白的非冗余大分子蛋白数据库，输出Evalue<1E-10的比对结果。如果一个非CRISPR相关蛋白数据库发现的同源蛋白小于100％，那么则说明这个家族的蛋白在CRISPR区域是富集的，通过这种方法我们对CRISPR富集大分子蛋白质家族进行鉴定。
6、蛋白功能和结构域的注释：利用Pfam数据库，NR数据库以及从NCBI收集的Cas蛋白对CRISPR富集大分子蛋白质家族进行注释，得到新的CRISPR/Cas蛋白质家族。利用Mafft对每个CRISPR/Cas家族蛋白进行多重序列比对，然后用JPred和HHpred进行保守结构域分析，鉴定含有RuvC结构域的蛋白质家族。
在此基础上，本发明人获得了一种全新的Cas效应蛋白，即Cas12g，其包括2种活性同源物序列，分别命名为Cas12g.1(SEQ ID NO:1)、Cas12g.2(SEQ ID NO:2)，两种同源物的编码DNA分别如SEQ ID NOs:19、20所示。Cas12g.1、Cas12g.2所对应的原型同向重复序列(pre-crRNA中所含有的repeat序列)分别如SEQ ID NOs:37、38所示。
实施例2.Cas12h基因和Cas12h导向RNA的获得
1、CRISPR和基因的注释：使用Prodigal对将NCBI和JGI数据库的微生物基因组和宏基因组数据进行基因注释得到所有蛋白，同时用Piler-CR进行CRISPR座的注释，参数均为默认参数。
2、蛋白质的过滤：通过序列一致性对注释蛋白去冗余，去除序列完全一致的蛋白，同时将长度大于800个氨基酸的蛋白划分为大分子蛋白。由于目前发现的所有第二类CRISPR/Cas***的效应蛋白长度多大于900个氨基酸，所以为了降低计算复杂度，我们在挖掘CRISPR效应蛋白的时候只对大分子蛋白进行考虑。
3、CRISPR相关大分子蛋白的获得：将每一个CRISPR座上下游延伸10Kb，将对CRISPR邻近区间内的非冗余大分子蛋白进行鉴定。
4、CRISPR相关大分子蛋白质的聚类：使用BLASTP对非冗余大分子CRISPR相关蛋白进行内部的两两比对，输出Evalue<1E-10的比对结果。使用MCL对BLASTP的输出结果进行聚类分析，CRISPR相关蛋白质家族。
5、CRISPR富集大分子蛋白质家族的鉴定：使用BLASTP对CRISPR相关蛋白质家族的蛋白比对到去除去CRISPR相关蛋白的非冗余大分子蛋白数据库，输出Evalue<1E-10的比对结果。如果一个非CRISPR相关蛋白数据库发现的同源蛋白小于100％，那么则说明这个家族的蛋白在CRISPR区域是富集的，通过这种方法我们对CRISPR富集大分子蛋白质家族进行鉴定。
6、蛋白功能和结构域的注释：利用Pfam数据库，NR数据库以及从N CBI收集的Cas蛋白对CRISPR富集大分子蛋白质家族进行注释，得到新的CRISPR/Cas蛋白质家族。利用Mafft对每个CRISPR/Cas家族蛋白进行多重序列比对，然后用JPred和HHpred进行保守结构域分析，鉴定含有Ruv C结构域的蛋白质家族。
在此基础上，本发明人获得了一种全新的Cas效应蛋白，即Cas12h，其包括2种活性同源物序列，分别命名为Cas12h.1(SEQ ID NO:3)、Cas 12h.2(SEQ ID NO:4)，两种同源物的编码DNA分别如SEQ ID NOs:21、22所示。Cas12h.1、Cas12h.2所对应的原型同向重复序列(pre-crRNA中所含有的repeat序列)分别如SEQ ID NOs:39、40所示。
实施例3.Cas12h.1蛋白的PAM结构域鉴定
1.构建重组质粒pACYC-Duet-1+CRISPR/Cas12h.1并测序。根据测序结果，对重组质粒pACYC-Duet-1+CRISPR/Cas12h.1进行结构描述如下：将载体pACYC-Duet-1的限制性内切酶Pml I和Kpn I识别序列间的小片段替换为SEQ ID NO:21所示的序列中自5’末端起第1至2664位所示的双链DNA分子。重组质粒pACYC-Duet-1+CRISPR/Cas12h表达SEQ ID NO:3所示的Cas12h.1蛋白和SEQ ID NO:39所示的Cas12h.1导向RNA。
2.重组质粒pACYC-Duet-1+CRISPR/Cas12h.1中含有表达盒，该表达盒的核苷酸序列如SEQ ID NO:94所示。SEQ ID NO:94所示的序列中，自5’末端起第1至44位为pLacZ启动子的核苷酸序列，第45至2708位为Cas12h.1基因的核苷酸序列，第2709至2794位为终止子的核苷酸序列(用于终止转录)。自5’末端起第2795至2834位为J23119启动子的核苷酸序列，第2835至3007位为CRISPR阵列的核苷酸序列，第3008至3093位为rrnB-T1终止子的核苷酸序列(用于终止转录)。
3.重组大肠杆菌的获得：将重组质粒pACYC-Duet-1+CRISPR/Cas12h.1导入大肠杆菌EC100中，得到重组大肠杆菌，命名为EC100/pACYC-Duet-1+CRISPR/Cas12h.1。将重组质粒pACYC-Duet-1导入大肠杆菌EC100中，得到重组农杆菌，命名为EC100/pACYC-Duet-1。
4.PAM文库的构建：人工合成SEQ ID NO:93所示的序列，并连接到pUC19载体，其中SEQ ID NO:93所示的序列包括5’端八个随机碱基和靶序列。对PAM文库的靶标序列5’端前面设计了8个随机碱基构建质粒文库。将质粒分别转入到含有Cas12h.1基因座的大肠杆菌中和不含有Cas.12h.1基因座的大肠杆菌中。在37℃下处理1小时后，我们对质粒进行提取，并对PAM区域序列进行PCR扩增和测序。
5.PAM文库结构域的获得：分别统计实验组和对照组中65,536种组合的PAM序列出现次数，并用各自组所有的PAM序列数目进行标准化。对于任意一条PAM序列，当log2(对照组标准化值/实验组标准化值)大于2时，认为这条PAM被显著消耗。用Weblogo对显著消耗的PAM序列进行预测，结果如图1所示，Cas12h.1的PAM结构域为5’-TG结构，这种PAM可以与其他PAM为5’-TTN的***相互补充，扩大识别范围。
尽管本发明的具体实施方式已经得到详细的描述，但本领域技术人员将理解：根据已经公布的所有教导，可以对细节进行各种修改和变动，并且这些改变均在本发明的保护范围之内。本发明的全部分为由所附权利要求及其任何等同物给出。
SEQUENCE LISTING
<110> 中国农业大学
<120> CRISPR/Cas效应蛋白及***
<130> IDC200414
<150> CN 201810426666.1
<151> 2018-05-07
<160> 112
<170> PatentIn version 3.5
<210> 1
<211> 882
<212> PRT
<213> 人工序列
<220>
<223> Cas12g.1的氨基酸序列
<400> 1
Met Leu Tyr Thr Met Asn Val Lys Thr Ile Lys Leu Lys Val Asp Ala
1 5 10 15
Thr Lys Glu Val Glu Ser Arg Leu Thr Lys Met Leu Leu Val His Asn
20 25 30
Asn Ile Gly Arg Glu Ile Ile Asn Phe Leu Ile Leu Cys Ser Gly Asn
35 40 45
Asp Asn Ile Arg Lys Thr Lys Phe Asp Glu Phe Gly Asn Ser Tyr Asp
50 55 60
Glu Phe Cys Asn Leu Lys Leu Asp Gln Phe Asn Leu Tyr Asp Arg Leu
65 70 75 80
Thr Glu Ile His Asp Glu Val Thr Leu Glu Asp Phe Gln Lys Thr Leu
85 90 95
Asn Asp Ile Tyr Asp Leu Val Leu Asn Ser Lys Ser Phe Ser Asn Val
100 105 110
Ser Ser Thr Ile Phe Asn Lys Asn Lys Lys Val Asn Phe Asp Glu Thr
115 120 125
Lys Lys Gly Asp Leu Ser Arg Lys Cys Leu Met Asn Ala Arg Asp Trp
130 135 140
Gly Val Leu Pro Leu Ile Ser Val Asp Asp Asp Ile Val Thr Cys Gly
145 150 155 160
Thr Leu Lys Gly Ile Leu Ser Glu Cys Gln Ser Arg Ile Leu Ser Trp
165 170 175
Asn Glu Cys Asn Leu Ser Thr Lys Glu Thr Tyr Ser Glu Lys Lys Ser
180 185 190
Glu Tyr Gln Ser Ile Leu Asp Asp Ser Met Thr Lys Asp Ala Asp Val
195 200 205
Thr Thr Ala Met Ile Gln Phe Met Asp Asp Val Ser Asn Val Tyr Gly
210 215 220
Ser Asn Asn Glu Asn Gln Leu Lys Trp Phe Asn Asn Arg Phe Leu Thr
225 230 235 240
Tyr Val Arg Asn Lys Ile Arg Pro Phe Leu Leu Thr Asn Ser Pro Ile
245 250 255
Asp Asn Phe Glu Gln Ser Asp Thr Ser Tyr Asn Cys Ser Ile Glu Ile
260 265 270
Val Arg Ile Leu Ser Lys Tyr Glu Ile Leu Trp Lys Asp Glu Val Ser
275 280 285
Val Asn Arg Tyr Lys Lys Thr Cys Asp Asp Gly Ile Asn Ile Glu Lys
290 295 300
Tyr Arg Tyr Leu Val His Ala Lys Ser Asp Phe Leu Arg Tyr Lys Glu
305 310 315 320
Thr Ala Ser Phe Lys Glu Ile His Ala Val Lys Ser Pro Ile Ser Leu
325 330 335
Cys Phe Gly Asn Asn Tyr Gln Pro Phe Ser Leu Ser Asp Val Gly Asp
340 345 350
Arg His Asn Ile Asn Phe Gly Tyr Lys Phe Gly Lys Leu Gly Lys Gln
355 360 365
Arg Lys Glu Cys Ser Phe Asn Leu Asn Tyr Arg Arg Lys Lys Val Lys
370 375 380
Tyr Ala Asn Thr Pro Val Arg Ser Asp Glu Asn Lys Cys Tyr Leu Asp
385 390 395 400
Asn Leu Glu Ile Glu Asp Ala Lys Asn Gly Ser Tyr Lys Leu Ser Tyr
405 410 415
Met Val Asn Lys Lys Tyr Lys Arg Glu Ser Phe Ile Lys Glu Pro Lys
420 425 430
Met Lys Met Tyr Asn Gly Lys Leu Tyr Met Tyr Phe Pro Met Ser Asn
435 440 445
Glu Phe Glu Glu Asp Arg Asp Ser Phe Ala Leu Leu Thr Tyr Phe Ser
450 455 460
Arg Ser Ser Asn Ser Lys Ser Gln Ile Asp Glu Ala Ser Asn Ile Leu
465 470 475 480
Gln Asn Arg Lys Ile Arg Val Cys Gly Val Asp Leu Gly Ile Asn Pro
485 490 495
Thr Phe Ala Leu Ser Val Leu Glu Tyr Ser Asp Asn Lys Ile Thr Asp
500 505 510
Thr Asn Ile Gly Met Lys His Glu Gly Ser Tyr Asn Asn Phe Ser Glu
515 520 525
Ile Arg Lys Gln Ile Asn Asp Val Thr Asp Met Ile Ser Tyr Leu Lys
530 535 540
Ser Lys Tyr Asp Asn Cys Glu Lys Asp Tyr Ser Ser Lys Ile Asp Asp
545 550 555 560
His Ile Lys Ser Arg Leu Asn Glu Glu Ile Ser Asn Phe Cys Asp Leu
565 570 575
Val Ser Tyr Lys Arg Asn Lys Asn Thr Ile Ile Arg Lys Glu Ile Lys
580 585 590
Asn Val Glu Lys Glu Ile Asn Lys Ile Lys Asn Cys Arg Arg His Thr
595 600 605
Leu Lys Lys Asp Leu Thr Glu Asn Phe Gly Trp Val Ser Ala Leu Asn
610 615 620
Glu Phe Ile Ser Leu Lys His Ser Phe Asn Asp Met Gly Glu Ser Phe
625 630 635 640
Asp Ser Lys Thr Asn Pro Ser Tyr Ser Tyr Phe Glu Lys Trp Lys Arg
645 650 655
Tyr Ile Asp Asn Ile Lys Asp Asp Ser Leu Lys Thr Val Ser Arg Glu
660 665 670
Ile Leu Asn Phe Cys Ile Glu Asn Ser Val Asp Phe Ile Ala Leu Glu
675 680 685
Asp Leu Gln Thr Phe Ala Pro Ser Asp Asp Arg Thr Lys Ser His Asn
690 695 700
Lys Leu Thr Gln Leu Trp Cys Phe Gly Lys Leu Lys Lys Cys Leu Glu
705 710 715 720
Asp Ile Ala Ser Met Tyr Gly Ile His Val Tyr Ser Ser Thr Asp Pro
725 730 735
Arg Asn Thr Ser Asp Thr His Phe Glu Ser Lys Asn Phe Gly Tyr Arg
740 745 750
Asp Glu Ser Asn Lys His Asn Leu Trp Val Asn Val Asp Gly Glu Tyr
755 760 765
Thr Val Val Asp Ser Asp Ile Asn Ala Ser Lys Asn Ile Ala Asn Arg
770 775 780
Phe Leu Thr His His Lys Asp Leu Lys Gln Leu Pro Met Ile Gly Asp
785 790 795 800
Gly Thr Leu Phe Lys Ile Asp Ser Ser Ser Lys Arg Asn Lys Ser Phe
805 810 815
Ala Val Lys Leu Asn Ile His Lys Asn Val Tyr Glu Leu Ile Asp Gly
820 825 830
Glu Phe Val Lys Ser Asn Lys Lys Pro Asn Gly Thr Ser Arg Lys Gln
835 840 845
Thr Ala Tyr Ile His Gly Asp Met Phe Ile Asp Ser Ile Ser His Lys
850 855 860
Asn Lys Lys Met Phe Leu Arg Glu Asn Leu Ile Arg Asn Gly Phe Ile
865 870 875 880
Ser Lys
<210> 2
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<223> Cas12g.2的氨基酸序列
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Met Asn Lys Thr Asp Thr Gln Asn Asn Glu Gln Ile Asn Lys Pro Thr
1 5 10 15
Gln Leu Leu Asn Asn Lys Asp Ile Glu Leu Thr Val Lys Thr Val Lys
20 25 30
Ser Ala Thr Val Lys Val Asp Asn Asn Ser Lys Lys Glu Leu Phe Gly
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Leu Phe Asn Tyr Phe Thr Ser Val Ala Ser Gly Ile Lys Asp Lys Val
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Tyr Asn Leu Gln Ser Asp Glu Lys Thr Ala Pro Ile Phe Asn Asp Tyr
65 70 75 80
Val Lys Gln Pro Gln Arg Gly Arg Ser Ala Ala Thr Thr Leu Phe Thr
85 90 95
Lys Leu Asp Ala Glu Lys Thr Tyr Thr Ser Gln His Ser Phe Pro Gly
100 105 110
Lys Trp Arg Asp Ser Gly Ile Phe Pro Leu Tyr Asn Lys Glu Ser Glu
115 120 125
Lys Tyr Asp Leu Ser Thr His Gly Tyr His Tyr Ser Ala Asn Ala Glu
130 135 140
Ile His Thr Gln Leu Asp Ser His Asp Glu Cys Asn Lys Glu Cys Glu
145 150 155 160
Lys Glu Tyr Ala Ala Leu Arg Asp Glu Val Asn Asn Tyr Lys Tyr Glu
165 170 175
Phe Thr Leu Gln Phe Lys Ala Glu Asn Ala Glu Lys Phe Tyr Asn Phe
180 185 190
Val Glu Lys Leu Thr Leu Met Gly Trp Arg Tyr Asp Ala Thr Phe Arg
195 200 205
Ser Phe Phe Glu Leu His Met His Pro Lys Leu Lys Thr Gly Glu Thr
210 215 220
Thr Tyr Arg Ala Thr Tyr Lys Leu Pro Ser Gly Lys Ser Lys Arg Tyr
225 230 235 240
Ser Phe Phe Arg Asp Asp Ile Ala Asp Glu Ile Ala Lys Asn Pro Glu
245 250 255
Phe Trp Pro Met Leu Glu Ser Ser Asn Ala Ile Ser Trp Ile Asn Ser
260 265 270
Asn Asn Leu Leu Ser Arg Lys Lys Asp Lys Ala Asn Tyr Ser Ser Thr
275 280 285
Ser Leu Ile Lys Ser Gln Ile Arg Leu Tyr Leu Gly Asn Asn Gly Val
290 295 300
Pro Phe Thr Ala Arg Glu His Asp Gly Arg Ile Tyr Phe Ser Phe Arg
305 310 315 320
Leu Pro Ala Ile Asn Gly Glu Lys Gly Arg Met Val Glu Ile Pro Cys
325 330 335
Ser Tyr Lys Lys Val Phe Asn Gly Lys Ala Arg Lys Ser Cys Tyr Leu
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Gly Gly Leu Thr Ile Glu Lys Thr Asp Ala Gly Lys His Ile Phe Lys
355 360 365
Tyr Ser Val Asn Asn Lys Lys Pro Gln Val Ala Glu Leu Asn Glu Cys
370 375 380
Phe Leu Arg Leu Val Val Arg Asn Arg Glu Tyr Phe Asn Asn Val Val
385 390 395 400
Ala Gly Lys Ile Thr Asp Ile Asn Thr Asp His Phe Asp Phe Tyr Val
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Asp Leu Pro Leu Asn Val Lys Glu Asp Pro Ile His Asp Leu Ser Ser
420 425 430
Thr Glu Val Phe Gly Lys Asn Gly Leu Arg Ser Tyr Tyr Ser Ser Ala
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Tyr Pro Glu Ile Lys Asn Leu Gly Ser Gln Ile Glu Thr Gly Lys Asn
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Leu Thr Cys Pro Ile Thr Lys Thr His Asn Ile Met Gly Ile Asp Leu
465 470 475 480
Gly Gln Arg Asn Pro Phe Ala Tyr Cys Ile Lys Asp Asn Thr Gly Lys
485 490 495
Leu Ile Ala Gln Gly His Met Asp Gly Ser Lys Asn Glu Thr Tyr Lys
500 505 510
Lys Tyr Ile Asn Phe Gly Lys Glu Ser Thr Ser Val Ser His Leu Ile
515 520 525
Lys Glu Thr Arg Ser Tyr Leu His Gly Asp Pro Glu Ala Ile Ser Lys
530 535 540
Glu Leu Tyr Asn Glu Val Ala Gly Phe Cys Asn Asn Pro Val Ser Tyr
545 550 555 560
Glu Glu Tyr Leu Lys Tyr Leu Asp Ser Lys Lys Phe Leu Ile Asn Lys
565 570 575
Glu Asp Leu Ser Lys Asn Ala Met His Leu Leu Arg Gln Lys Asp His
580 585 590
Asn Trp Ile Gly Arg Asp Trp Leu Trp Tyr Ile Ser Lys Gln Tyr Lys
595 600 605
Lys His Asn Glu Asn Arg Met Gln Asp Ala Asp Trp Arg Gln Thr Leu
610 615 620
Tyr Trp Ile Asp Ser Leu Tyr Arg Tyr Ile Asp Val Met Lys Ser Phe
625 630 635 640
His Asn Phe Gly Ser Phe Tyr Asp Lys Asn Leu Lys Lys Lys Val Asn
645 650 655
Gly Thr Val Val Gly Phe Cys Lys Thr Val His Asp Gln Ile Asn Asn
660 665 670
Asn Asn Asp Asp Met Phe Lys Lys Phe Thr Asn Glu Leu Met Ser Val
675 680 685
Ile Arg Glu His Lys Val Ser Val Val Ala Leu Glu Lys Met Asp Ser
690 695 700
Met Leu Gly Asp Lys Ser Arg His Thr Phe Glu Asn Arg Asn Tyr Asn
705 710 715 720
Leu Trp Pro Val Gly Gln Leu Lys Thr Phe Met Glu Gly Lys Leu Glu
725 730 735
Ser Phe Asn Val Ala Leu Ile Glu Ile Asp Glu Arg Asn Thr Ser Gln
740 745 750
Val Cys Lys Glu Asn Trp Ser Tyr Arg Glu Ala Asp Asp Leu Tyr Tyr
755 760 765
Val Thr Asp Gly Glu Ser His Lys Val His Ala Asp Glu Asn Ala Ala
770 775 780
Asn Asn Ile Val Asp Arg Cys Ile Ser Arg His Thr Asn Met Phe Ser
785 790 795 800
Leu His Met Val Asn Pro Lys Asp Asp Tyr Tyr Val Pro Thr Cys Ile
805 810 815
Trp Asp Thr Thr Glu Glu Ser Gly Lys Arg Val Arg Gly Phe Leu Thr
820 825 830
Lys Leu Tyr Lys Asn Ser Asp Val Val Phe Thr Lys Lys Gly Asp Lys
835 840 845
Leu Val Lys Ser Lys Thr Ser Val Lys Glu Leu Lys Lys Leu Val Gly
850 855 860
Lys Thr Lys Glu Lys Arg Gly Gln Tyr Trp Tyr Arg Phe Glu Gly Lys
865 870 875 880
Ser Trp Ile Asn Glu Ala Asp Arg Asp Thr Ile Ile Leu Asn Ala Lys
885 890 895
Lys Ile Ser Arg Glu Arg Asp Asn Gly Glu Gln Ser Thr Asp Thr Arg
900 905 910
Ser Gln Asn Val Thr Val Ser Val Leu Asp Val Cys Glu Thr Ala Glu
915 920 925
Lys Lys Lys Leu Val Leu Val
930 935
<210> 3
<211> 887
<212> PRT
<213> 人工序列
<220>
<223> Cas12h.1的氨基酸序列
<400> 3
Met Ala Leu Ile Gln Arg Ala Gly Val Leu Lys Thr Lys Ser Asp Phe
1 5 10 15
Pro Lys Val Ile Lys Asp Trp His Asp Ser Leu Leu Ala Asp Tyr Arg
20 25 30
Lys Phe Phe Pro Ile Ile Phe Ser Trp Cys Pro Glu Tyr Gly Tyr Thr
35 40 45
Thr Ile Gln Asp Asn Lys Pro Val Phe Val Ser Pro Glu Glu Arg Met
50 55 60
Glu Ser Ile Arg Lys Glu Ala Lys Glu His Leu Asn Glu Val Leu Ala
65 70 75 80
Phe Gly Lys Met Ile Gly Ser Lys Gly Val Gly Gly Ser Ser Ser Tyr
85 90 95
Ala Ile Phe Tyr Lys His His Lys Asn Asn Glu Asn Gly Ala Tyr Thr
100 105 110
Pro Ser Arg Ala Lys Phe Met Lys Glu Gly Ile His Asn Arg Arg Val
115 120 125
Glu Leu Val Asp Val Leu Met Leu Asn Ala Ile Pro Asp Glu Glu Trp
130 135 140
Val Lys Ile Ala Gln Glu Val Val Gly Tyr Ser Glu Glu Arg Leu Lys
145 150 155 160
Leu Tyr Trp Asn Lys Phe Ile Ala Lys Arg Val Val Ser His Asp Arg
165 170 175
Lys Leu Gly Lys Ile Val Arg Glu Lys Tyr Leu Glu Pro Lys Gly Leu
180 185 190
Val Cys Ala Gln Pro Glu Asn Ser Thr Tyr Cys Arg Val Leu Thr Glu
195 200 205
Ile Ile Lys Arg Gln Leu His Ser Gln Ile Glu Lys Ser Lys Phe His
210 215 220
Glu Glu Glu Leu Lys Ser Ile Glu Lys Thr Val Ser Glu Phe Asp Ser
225 230 235 240
Pro Leu Leu Asp Phe Ile Cys Gln Tyr Ala Glu Glu Leu Asn Gln Ile
245 250 255
Asn Ser Gly Leu Ser Lys Tyr Val Ile Lys Asn Ala Val Lys Glu Val
260 265 270
Ile Ser Pro Pro Glu Lys Gln Ser Glu Ile Tyr Val Gln Ser Gln Val
275 280 285
Leu Ser Gln Glu Lys Tyr Lys Pro Leu Val Asn Ala Thr Ile Lys Glu
290 295 300
Ile Leu Ser Gly Tyr Glu Gln Trp Lys Val Lys Ser Arg Tyr Glu Asn
305 310 315 320
Arg Leu Lys Asn Arg Lys Tyr Val Leu Tyr Pro Lys Leu Ser Ala Asn
325 330 335
Tyr Lys Ile Pro Ile Gly Gln Asn Ser Leu Gly Lys Phe Lys Ile Asn
340 345 350
Val Ser Glu Asn Gly Glu Ile Val Ile Arg Leu Asn Asp Met Ala Asp
355 360 365
Val Val Cys Met Pro Ser Lys Tyr Phe Phe Asn Leu Lys Ser Ser Pro
370 375 380
Val Val Asp Lys Lys Lys Gln Leu Val Gly Tyr Gln Ile Ser Phe Asn
385 390 395 400
His Asn Ser Arg Arg Lys Glu Pro Thr Glu Lys Pro Asp Phe Asn Gly
405 410 415
Ile Val Lys Glu Ile Gly Leu Gln Leu Lys Asp Asp Gly Arg Phe Tyr
420 425 430
Ile Thr Leu Pro Tyr Cys Met Glu Tyr Ser Asn Asp Asn Phe Asp Leu
435 440 445
Ile Arg Pro Leu Leu Thr Ser Ser Pro Thr Glu Asp Gln Ile Lys Lys
450 455 460
Met Pro Ser Glu Phe Asn Val Val Gly Phe Asp Leu Asn Leu Ser Met
465 470 475 480
Pro Leu Pro Ile Thr Arg Ala Ile Val Gly Lys Ser Val Lys Gly Glu
485 490 495
Ile Asn Val Glu Tyr Leu Gly Gln Ala Lys Val Ile Glu Ser Thr His
500 505 510
Leu Ile Tyr Asp Asn Asn Arg Cys Lys Val Leu Ile Ala Tyr Lys Arg
515 520 525
Gln Cys Asp Leu Ile Lys Arg Ala Ile Arg Glu Trp Lys Ile Cys Lys
530 535 540
Gly Lys Asn Ile Asp Ile Ser Glu Lys Thr Tyr Glu Trp Leu Glu Ser
545 550 555 560
His Thr Lys Arg Trp Asn Pro Ser Arg Gln Pro Glu Ser Met Gln Asp
565 570 575
Arg Phe Ser Val Ser Lys Met Arg Ile Gln Ile Leu Val Asn Lys Ala
580 585 590
Lys Ser Arg Ile Ala Lys Tyr Asn Asp Asn Ser Trp Lys Thr Gly His
595 600 605
Gly Asn Glu Ser Glu Leu Ile Arg Leu Ile Asp Ala Asp Asp Ala Tyr
610 615 620
Asn Ser Leu Val Ser Thr Tyr Asn Arg Ile His Leu Lys Ser Asn Gln
625 630 635 640
Phe Ile Tyr Ala Leu Pro Ser Lys Asn Asn Ser Arg Ser Asn Lys Lys
645 650 655
Glu Tyr Cys Leu Arg Arg Ile Ala Ala Lys Ile Ala Arg Tyr Cys His
660 665 670
Leu His Asn Val Asn Ile Cys Ile Gly Glu Asn Leu Ser Phe Gln Gln
675 680 685
Asp Ser Asp Asn Ile Ser Lys Asp Asn Ser Leu Val Arg Leu Phe Ser
690 695 700
Ser Lys Ser Ile Ala Asn Tyr Met Lys Leu Ala Met Glu Lys Phe Gly
705 710 715 720
Ile Ala Phe Ile Asp Ser Ala Asp Pro Ser Gly Thr Ser Lys Thr Asp
725 730 735
Pro Val Thr Gly Asn Ile Gly Tyr Arg Asn Lys Phe Asp Lys Arg Lys
740 745 750
Leu His Val Ile Arg Asn Gly Asn Trp Gly Trp Val Asp Ser Asp Ile
755 760 765
Ala Ala Ser Leu Asn Ile Leu Ile Arg Gly Ile Asn Arg Ser Ile Val
770 775 780
Pro Tyr Lys Phe Phe Val Gly Lys Lys Lys Gln Glu Ser Lys Arg Leu
785 790 795 800
Asn His Phe Leu Asn Lys Ile Phe Gly Thr Thr Lys Val Phe Phe Tyr
805 810 815
Glu Asp Gln Phe Gly Phe Ala Asn Pro Ser Leu Ser Lys Lys Glu Gly
820 825 830
Glu Asn Leu Ile Ala Asn Gln Tyr Leu Tyr Tyr Arg Glu Gly Lys Phe
835 840 845
Val Thr Gln Lys Ile His Arg Gln Ile Glu Asp Asp Phe Lys Lys Ile
850 855 860
Asp Phe Ser Asn Thr Pro Glu Val Asn Leu Ile Pro Ser Gly Val Lys
865 870 875 880
Leu Lys Asn Phe Gln Phe Glu
885
<210> 4
<211> 921
<212> PRT
<213> 人工序列
<220>
<223> Cas12h.2的氨基酸序列
<400> 4
Met Ala Thr Arg Ser Phe Ile Arg Thr Gly Asn Leu Lys Ala Lys Asn
1 5 10 15
Thr Ala Glu Glu Val Met Gln Trp Tyr Ala Asp Leu Gln Ser Asp Tyr
20 25 30
Arg Ser Phe Leu Asn Leu Phe Phe Gly Trp Met Ala Ile Gly Tyr Gly
35 40 45
Thr Asn Ala Glu Asp Glu Val Phe Tyr Thr Ser Lys Glu Glu Ser Glu
50 55 60
Arg Leu Arg Ser Leu Thr Ile Gly Asp Ala Lys Lys Glu Gln Leu Ala
65 70 75 80
Val Ser Phe Ile Glu Leu Leu Leu Lys Gly Gly Glu Asn Ala Ser Ser
85 90 95
Cys Tyr Asn Val Phe Tyr Arg Asn Tyr Lys Ser Leu Gly Lys Ala Lys
100 105 110
Leu Thr Gln Lys Lys Asn Asp Phe Leu Ser Ala Leu Pro Leu Leu Asp
115 120 125
Glu Asn Lys Ile Lys Glu Tyr Phe Lys Thr Asp Glu Gln Leu Ser Gln
130 135 140
Ile Cys Ile Glu Glu Trp Leu Glu Tyr Gly Val Lys Asn Leu Pro Leu
145 150 155 160
Pro Glu Ile Trp Ala Glu Val Ser Pro Arg Leu Ala Ser Ile Glu Arg
165 170 175
Ser Leu Gly Val Asp Leu Arg Leu Ala Phe Gly Leu Ser Cys Ile Arg
180 185 190
Ser Arg Asp Cys Asn Tyr Cys Arg Ile Leu Ile Glu Met Val Gly Arg
195 200 205
Asp Leu Arg Ser Ile Phe Glu Lys Tyr Asn Asn His Leu Leu Glu Thr
210 215 220
Glu Lys Ile Lys Leu Ser Met Asn Asp Lys Gln Gly Pro Val Tyr Asp
225 230 235 240
Ser Ile Cys Cys Phe Ala Ala Glu Leu Glu Ser Lys Asn Ser Gly Leu
245 250 255
Thr Lys Tyr Val Leu Thr Lys Gly Ile Asp His Val Lys Lys Gly Thr
260 265 270
Gly Glu Lys Thr Asp Ile Arg Leu Ala Val Lys Glu Leu Lys Lys Asn
275 280 285
Lys Tyr Arg Ile Leu Ile Glu Ser Ser Tyr Ser Glu Ile Met Ser Ala
290 295 300
Tyr Ser Cys Trp Arg Thr Lys Lys Gln Leu Glu Lys Arg Lys Leu Tyr
305 310 315 320
Pro Cys Phe Asp Pro Asn Arg Asn Asp Tyr Lys Val Pro Val Gly Gln
325 330 335
Gly Ser Leu Gly Asn Phe Thr Val Ser Val Glu Asp Ser Gly Asp Val
340 345 350
Leu Ile Glu Ile Val Gly Val Gly Val Ile Arg Cys Ala Ala Ser Cys
355 360 365
Tyr Phe Ser Gly Ile Val Phe Asp Glu Ile Arg Asn Lys Asn Gly Arg
370 375 380
Thr Gly Tyr Ser Leu Asn Phe Cys His Lys Ser Ile Ser Lys Gly Lys
385 390 395 400
Lys Ala Val Lys Ala Ala Ser His Thr Gly Asp Lys Ile Ser Gly Val
405 410 415
Leu Lys Glu Ile Gly Leu Arg Asn Thr Asp Ser Gly Phe Phe Val Ser
420 425 430
Leu Pro Tyr Ser Ile His His Asp Glu Lys Asn Phe Lys Ile Ala Glu
435 440 445
Phe Phe Met Ser Ala Cys Pro Lys Lys Glu Asn Val Glu Asn Leu Pro
450 455 460
Asp Lys Ile Val Val Gly Ala Ile Asp Leu Asn Val Ser Asn Pro Val
465 470 475 480
Ala Ala Val Lys Ala Val Val Tyr Arg Asp Asp Lys Ser Gly Gln Leu
485 490 495
Asn Ala Leu Asp Tyr Gly Ser Gly Asn Leu Ile Lys Lys Pro Phe Met
500 505 510
Leu Val Ala Asn Gly Pro Arg Ile Lys Asn Leu Ile Glu Ile Arg Asp
515 520 525
Asp Ala Arg Arg Val Ile Gly Ala Ile Arg Glu Phe Lys Val Ser Asn
530 535 540
Ala Val Lys Glu His Val Gly Glu Asp Thr Arg Asp Phe Leu Ile Leu
545 550 555 560
Cys Gly Asp Thr Lys Ser Ser Ser Thr Arg Tyr Leu Ile Gln Ser Trp
565 570 575
Val Lys Lys Ile Asn Ser Arg Leu Arg Lys Ile Lys Phe Glu Met Arg
580 585 590
Ser Gly Gly Tyr Arg Asp Cys Ala Asp Asn Ile Arg Leu Ile Glu Ala
595 600 605
Met Asp Gln Cys Ala Ser Met Ala Glu Ser Tyr Asn Arg Ile His Leu
610 615 620
Lys Ser Gly Glu Lys Leu Val Lys Val Ala Lys Phe Asp Lys Ser Arg
625 630 635 640
Ala Asn Phe Arg Asn Phe Val Leu Arg Gln Leu Ala Ser Lys Ile Ala
645 650 655
Asn Glu Met Lys Asp Cys Asn Val Val Phe Gly Glu Asp Leu Asp Phe
660 665 670
Ile Phe Asp Ser Asp Lys Asn Asn Asn Ala Leu Leu Arg Leu Phe Ser
675 680 685
Ala Ala Thr Leu Leu Lys Tyr Ile Ile Glu Ala Leu Glu Lys Ile Gly
690 695 700
Val Gly Phe Val Lys Val Ala Lys Asn Gly Thr Ser Gln Ser Asp Pro
705 710 715 720
Val Thr Ser Asn Pro Gly Trp Arg Asp Asp Lys Asn Lys Ser Arg Leu
725 730 735
Tyr Val Val Arg Asp Lys Gln Leu Gly Trp Ile Asp Ser Asp Leu Ala
740 745 750
Ala Thr Met Asn Ile Leu Ile Gln Gly Leu Asn His Ser Val Cys Pro
755 760 765
Tyr Lys Phe Tyr Val Lys Glu Tyr Glu Asn Lys Pro Asn Ser Thr Gln
770 775 780
Asp Ser Ile Asn Ala Ile Lys Lys Pro Glu Glu Ala Ile Gly Lys Arg
785 790 795 800
Ile Lys Arg Phe Phe Asn Leu Lys Tyr Gly Ser Ser Val Pro Lys Phe
805 810 815
Val Ser Asp Asp Arg Gly Arg Val Thr Phe Ala Lys Lys Ile Asp Ser
820 825 830
Thr Gln Thr Arg Leu Ile Asn Gln Phe Val Tyr Ala His Ser Ser Cys
835 840 845
Ile Val Thr Cys Glu Leu His Asn Glu Met Val Asn Lys Ile Lys Gln
850 855 860
Leu Ala Val Glu Lys Pro Asn Cys Gln Glu Phe Asp Val Thr Cys Asp
865 870 875 880
Pro Asp Gly Arg Tyr Asn Asn Phe Ala Leu Pro Glu Val His Asp Ser
885 890 895
Ser Lys Asp Val Gly Ala Lys Ala Leu Thr Thr Lys Asp Val Asp Phe
900 905 910
Lys Thr Ile Leu Lys Asp His Thr Ala
915 920
<210> 5
<211> 1297
<212> PRT
<213> 人工序列
<220>
<223> Cas12w.1的氨基酸序列
<400> 5
Met Gly Lys Asn Glu Asn Lys Tyr Gln Leu Ser Lys Thr Leu Arg Phe
1 5 10 15
Gly Leu Thr Leu Lys Glu Lys Ile Ser Asn Asn Glu Lys Thr Pro Tyr
20 25 30
Gln Ser His Ser Gln Phe Arg Asp Leu Ile Ile Leu Ser Glu Asn Arg
35 40 45
Ile Arg Glu Gly Ile Ser Thr Pro Gln Asn Arg Asp Leu Pro Ser Phe
50 55 60
Ile His Arg Ile Gln Asn Cys Thr Asp Phe Ile Asn Asp Phe Ile His
65 70 75 80
Asp Trp Trp Met Ile Leu Met His Thr Gly Gln Ile Glu Leu Asp Lys
85 90 95
Asp Tyr Tyr Lys Ser Leu Thr Lys Lys Val Gly Phe Val Gly Phe Trp
100 105 110
Tyr Lys Glu Asn Lys Lys Lys Gly Gly Lys Thr Lys Gln Pro Gln Ala
115 120 125
Arg Asn Ile Pro Met Gly Glu Leu Arg His Leu Cys Pro Gln Asn Thr
130 135 140
Lys Glu Cys Ala Thr Tyr Ile Thr Asp Tyr Trp Lys Asp Leu Leu Ile
145 150 155 160
Thr Ala Thr Asn Lys Leu Tyr Glu Ser Ser Glu Gln Gln Lys Lys Phe
165 170 175
Ile Lys Ala Met Glu Gln Asn Arg Thr Asp Asn Lys Pro Asn Glu Ile
180 185 190
Asp Leu Lys Lys Ser Phe Leu Ser Leu Val Ser Val Thr Met Glu Leu
195 200 205
Leu Asn Pro Ile Leu Asn Gly Gln Ile Leu Phe Asn Lys Met Asp Arg
210 215 220
Leu Asp Met Ser Lys Lys Ser Asp Asn Asp Phe Ile Asp Phe Val Asn
225 230 235 240
Asp His Glu Thr Val Arg Glu Leu Asn Asn Asp Ile Glu Glu Ile Ile
245 250 255
Ala Asp Phe Lys Glu Asn Gly Asn Asn Val Asn Tyr Cys Lys Ala Thr
260 265 270
Leu Asn Pro Asp Thr Ala Leu Lys Gln His Asn Asn Asn Ile Pro Asn
275 280 285
Asp Ile Ala Thr Asp Leu Glu Glu Leu Met Met Asp Ser Ile Val Gly
290 295 300
Asn Tyr Asp Asp Val Asn Ser Phe Met Asp Asn Tyr Val Ser Asn Leu
305 310 315 320
Ser Ala Lys Asp Lys Ile Lys Lys Ile Lys Asp Ser Asn Ile Ser Leu
325 330 335
Ile Tyr Arg Ala Ile Leu Phe Lys Tyr Lys Met Ile Pro Ala Asn Val
340 345 350
Arg Arg Asp Ile Ala Gln Gly Met Ala Lys Lys Leu Asn Lys Asp Glu
355 360 365
Glu Asn Ile Tyr Ser Phe Leu Cys Glu Phe Gly Thr Leu Arg Thr Pro
370 375 380
Gln Lys Asp Tyr Ala Asp Leu Lys Asp Lys Asp Ser Phe Asn Leu Asp
385 390 395 400
Asn Tyr Pro Leu Lys Val Ala Phe Asp Phe Ala Trp Glu Gly Leu Ala
405 410 415
Lys Ala Trp Tyr His Asp Gln Ser Asp Phe Pro Ile Asp Pro Cys Arg
420 425 430
Asp Phe Leu Gln Glu Asn Phe Asp Val Asn Leu Glu Glu Asp Gln Glu
435 440 445
Asp Glu Tyr Phe Leu Leu Tyr Ala Asp Leu Ile Glu Leu Asn Ala Leu
450 455 460
Leu Ser Thr Leu Asp Lys Gly Asn Pro Ala Asp Pro Asp Ser Ile Lys
465 470 475 480
Asn Glu Ala Leu Glu Met Val Glu Tyr Ile Asn Trp Asn Ser Leu Asp
485 490 495
Lys Lys Asn Gly Asn Tyr Tyr Lys Lys Ile Ile Lys Asn Arg Leu Lys
500 505 510
Ser Ser Lys Gly Asn Glu Thr Tyr Glu Arg Ile Lys Lys Glu Ile Ser
515 520 525
Met Ser Arg Gly Arg Leu Lys Asn Lys Ile Glu Lys Tyr Asp Asp Leu
530 535 540
Thr Ser Gln Tyr Lys Arg Ile Ala Met Asp Leu Gly Lys Lys Phe Ala
545 550 555 560
Ser Leu Arg Asp Lys Ile Ile Ala Ala Asn Glu Asp Asn Lys Val Thr
565 570 575
His Tyr Ala Met Ile Leu Glu Asp Ser Asn Cys Asp Lys Tyr Leu Leu
580 585 590
Leu Gln Lys Val Ser Asn Asn Ile Tyr His Cys Met Ser Tyr Asp Ser
595 600 605
Ser Asp Pro Lys Ala Tyr Tyr Val Asp Ser Ile Thr Ser Ser Ala Ile
610 615 620
Ala Lys Met Ile Arg Lys Glu Thr Asn Pro Ser Lys Ile Arg Glu Tyr
625 630 635 640
Ala Glu Leu Glu Glu Lys Glu Arg Glu Arg Arg Asn Val Asp Asp Trp
645 650 655
Cys Arg Phe Ile Ser Lys Lys Glu Tyr Asp Arg Arg Tyr Gln Leu Asn
660 665 670
Ile Asn Asn Gly Leu Ser Phe Glu Ala Leu Lys Lys Glu Ile Asp Ser
675 680 685
Lys Ser Tyr Ile Leu Val Lys Lys Asn Ile Ser Val Asp Ser Ile Arg
690 695 700
Glu Leu Val Glu Asn Glu Gly Cys Leu Leu Phe Pro Ile Val Asn Lys
705 710 715 720
Asp Leu Thr Lys Glu Arg Lys Thr Thr Glu Asp Asn Gln Phe Thr Lys
725 730 735
Asp Trp Asn Met Ile Phe Ser Gly Ser Glu Thr Asn Trp Arg Leu Thr
740 745 750
Pro Glu Phe Arg Val Thr Tyr Arg Asn Pro Val Pro Gly Tyr Pro Asn
755 760 765
Asp Lys Phe Gly Ser Lys Arg Tyr Ser Arg Phe Gln Met Asn Ala His
770 775 780
Phe Val Cys Asp Phe Ile Pro Ser Ser Asn Ser Tyr Thr Ser Asn Arg
785 790 795 800
Glu Gln Ile Ala Ile Phe Lys Asp Glu Gly Glu Gln Lys Lys Arg Val
805 810 815
Glu Glu Phe Asn Arg Thr Leu Ser Asn Ile Asn Gln Lys Phe Tyr Val
820 825 830
Ile Gly Ile Asp Arg Gly Gln Lys Glu Leu Ala Thr Leu Cys Val Val
835 840 845
Asp Gln Asp Lys Lys Ile His Gly Asp Phe Lys Ile Tyr Thr Arg Lys
850 855 860
Phe Asn Ser Glu Arg Lys Gln Trp Glu His Tyr Ser Leu Glu Gly Glu
865 870 875 880
Lys Gly Thr Arg Asn Ile Leu Asp Leu Ser Asn Leu Arg Val Glu Thr
885 890 895
Thr Ile Ile Ile Asp Gly Lys Pro Glu Arg Arg Gln Val Leu Val Asp
900 905 910
Leu Ser Glu Val Leu Val Lys Asp Lys Glu Gly Asn Tyr Thr Lys Pro
915 920 925
Asn Lys Met Gln Ile Lys Met Gln Gln Met Ala Tyr Val Arg Lys Leu
930 935 940
Gln Phe Gln Met Gln Ala Asn Pro Thr Glu Val Leu Glu Trp Tyr Glu
945 950 955 960
Gln Asn Pro Thr Glu Glu Leu Ile Ile Lys Asn Leu Val Asp Lys Glu
965 970 975
Asn Gly Glu Lys Gly Leu Ile Ser Phe Tyr Gly Thr Ala Leu Val Glu
980 985 990
Leu Asp Gln Thr Leu Pro Val Ser Lys Ile Lys Glu Met Leu Glu Glu
995 1000 1005
Phe Lys Ile Leu Lys Gln Arg Glu Ser Lys Lys Glu Asn Val Gln
1010 1015 1020
Lys Glu Leu Asn Asn Leu Thr Gln Leu Glu Ala Val Asp Ser Leu
1025 1030 1035
Lys Ala Gly Ile Val Ala Asn Met Val Gly Val Ile Ser Tyr Ile
1040 1045 1050
Leu Lys Thr Leu Asp Tyr Asn Ala Tyr Ile Ser Leu Glu Asp Leu
1055 1060 1065
Ser Thr Val Gln Ser Ser Thr Glu Phe Ala Ser Gly Ile Ser Gly
1070 1075 1080
Ala Ile Thr Lys Met Ser Arg Glu Glu Gly Arg Arg Ile Asp Val
1085 1090 1095
Glu Lys Tyr Ala Gly Leu Gly Leu Tyr Asn Phe Phe Glu Met Gln
1100 1105 1110
Leu Leu Arg Lys Leu His Arg Ile Gln Thr Asp Asn Gly Asn Ile
1115 1120 1125
Leu His Leu Val Pro Ala Phe Arg Ala Gln Lys Asn Tyr Asp His
1130 1135 1140
Ile Met Val Gly Lys Glu Lys Ile Lys Asn Gln Phe Gly Ile Val
1145 1150 1155
Phe Phe Val Asp Ala Ala Ala Thr Ser Ile Lys Cys Pro Arg Cys
1160 1165 1170
Gly Ala Val Asn Glu Asp Lys Phe Asn Pro Asp Lys Gln Lys Tyr
1175 1180 1185
Pro Asp Ala Glu Lys Gly Pro Lys Leu Arg Asn Arg Lys Glu Gln
1190 1195 1200
Ser Gly Lys Lys Val Trp Val Thr Arg Asp Lys Glu Asp Asp Asp
1205 1210 1215
Arg Ile Lys Cys Tyr Cys Cys Gly Phe Asp Thr Lys Glu Lys Asn
1220 1225 1230
Glu Gly Asn Pro Phe Met Tyr Ile Lys Ser Gly Asp Asp Asn Ala
1235 1240 1245
Ala Tyr Leu Ile Ser Asp Leu Gly Val Glu Ser Tyr Arg Lys Ala
1250 1255 1260
Tyr Glu Leu Ala Ala Thr Val Val Glu Asp Arg Lys Lys Thr Leu
1265 1270 1275
Thr Asn Asn Leu Asn Gln Ser Asn Tyr Lys Ile Arg Phe Leu Trp
1280 1285 1290
His Thr Met Tyr
1295
<210> 6
<211> 1257
<212> PRT
<213> 人工序列
<220>
<223> Cas12w.2的氨基酸序列
<400> 6
Met Asp Ala Asp Lys Thr Thr Lys Ala Ile Asn Glu Tyr Gln Thr Gln
1 5 10 15
Lys Thr Ile Arg Phe Gly Leu Thr Ala Thr Asn Gln Asn Leu Tyr Ser
20 25 30
Glu Glu Ile Met Lys Leu Leu Asn Ile Ser Glu Glu Arg Ile Ile Lys
35 40 45
Glu Lys Val Lys Val Asn Asn Asp Thr Asp Lys Thr Asn Gln Leu Arg
50 55 60
Gly Cys Leu Val Gln Ile Lys Lys Tyr Leu Lys Thr Trp Glu Asn Ile
65 70 75 80
Tyr Ala Gln Ile Asp Phe Leu Ala Ile Thr Lys Asp Tyr Tyr Lys Val
85 90 95
Ile Ser Lys Lys Ala Arg Phe Asp Phe Asp Lys Gly Asn Gly Ser Glu
100 105 110
Ile Lys Leu Ser Ser Leu Gln Ser Thr His Asn Lys Lys Lys Arg Tyr
115 120 125
Gln Tyr Ile Ile Asp Phe Trp Lys Glu Asn Leu Arg Lys Thr Glu Asn
130 135 140
Leu Tyr Arg Lys Ser Asp Asp Leu Leu Lys Ile Phe Glu Glu Ala Lys
145 150 155 160
Asn Gln Asn Arg Asp Asp Lys Lys Leu Asn Lys Val Glu Leu Arg Lys
165 170 175
Thr Phe Leu Asn Leu Phe Thr Leu Val Asn Glu Ser Leu Lys Pro Leu
180 185 190
Ile Glu Gly Asn Leu Phe Ile Val Asn Asp Asp Lys Ile Asp Glu Lys
195 200 205
Asn Ser Lys His Asn Tyr Val Phe Tyr Phe Ile Ser Lys Thr Glu Glu
210 215 220
Arg Arg Leu Leu Tyr Asp Asn Ile Cys Thr Leu Gln Asp Tyr Phe Lys
225 230 235 240
Asn Asn Gly Gly Tyr Val Pro Phe Gly Arg Val Thr Leu Asn Lys Trp
245 250 255
Thr Ala Leu Gln Lys Phe Asn Asn Arg Asp Ile Glu Ile Asn Arg Ile
260 265 270
Ile Lys Glu Leu Lys Ile Asn Asn Ile Ser Thr Gln Lys Thr Asp Tyr
275 280 285
Lys Tyr Asn Asp Phe Thr Glu Asn Phe Lys Glu Lys Lys Asp Glu Asn
290 295 300
Gly Lys Val Val Lys Asn Ser Ala Gly Asn Ile Ile Trp Asp Leu Lys
305 310 315 320
Ala Asn Ala Lys Ser Val Ile Glu Ile Cys Gln Phe Phe Lys Tyr Lys
325 330 335
Lys Val Pro Ile Asn Ala Arg Leu Asn Leu Ala Lys Arg Leu Ile Lys
340 345 350
Asp Asn Lys Leu Lys Lys Glu Gln Glu Asn Thr Phe Leu Ser Glu Phe
355 360 365
Gly Val Leu Lys Thr Pro Ala Phe Asp Tyr Ala Arg Asp Lys Glu Asn
370 375 380
Phe Asn Leu Thr Asn Tyr Pro Leu Lys Val Ala Phe Asp Tyr Ala Trp
385 390 395 400
Glu Asn Cys Ala Lys Asp Lys Tyr Glu Lys Ile Pro Phe Pro Lys Glu
405 410 415
Gln Cys Glu Arg Tyr Leu Gln Thr Ala Phe Glu Ile Asp Ala Thr Lys
420 425 430
Asp Glu Asn Lys Lys Leu Ile Asp Thr His Leu Asn Lys Tyr Ala Asp
435 440 445
Leu Leu Gln Phe Lys Ile Leu Leu Glu Arg Phe Lys Ala Glu Phe His
450 455 460
Lys Thr Asn Glu Glu Thr Asn Lys Asn Asn Ile Gln Lys Leu Arg Asn
465 470 475 480
Val Phe Ser Gly Leu Asp Tyr His Gly Asp Asn Arg Leu Asn Lys Asn
485 490 495
Gln Ile Gln Lys Ala Ile Glu Ala Trp Phe Asp Asn Lys Glu Gln Asn
500 505 510
Ile Gly Lys Lys Lys Glu Asn Glu Lys Leu Leu Thr Glu Asn Glu Lys
515 520 525
Asn Asn Phe Ser Leu Ser Met Gln Ile Ile Gly Gln Glu Arg Gly Gly
530 535 540
Leu Lys Asn Gly Ile Pro Lys Tyr Lys Glu Leu Thr Glu Met Phe Lys
545 550 555 560
Val Cys Ala Ser Lys Phe Gly Lys Gln Phe Ala Asp Leu Arg Asp Tyr
565 570 575
Phe Asn Glu Ala Tyr Glu Val Asp Lys Ile Lys Tyr Arg Ala Trp Ile
580 585 590
Ile Glu Asp Asp Lys Lys Asn Arg Phe Val Leu Phe Val Asn Lys Glu
595 600 605
Lys Ala Phe Asp Leu Thr Ser Glu Glu Gly Asp Leu Trp Phe Tyr Glu
610 615 620
Val Lys Ser Leu Thr Ser Lys Ser Leu Val Lys Phe Ile Lys Asn Arg
625 630 635 640
Gly Ala Tyr Pro Asp Phe His Asp Val Lys Asn Ser Phe His Tyr Ser
645 650 655
Ser Ile Lys Lys Asp Trp Gln Asn Tyr Lys Asn Asp Pro Glu Phe Leu
660 665 670
Asp Lys Leu Lys Glu Cys Leu Lys Asn Ser Lys Ile Ala Lys Asp Gln
675 680 685
Lys Trp Ala Lys Phe Cys Trp Asp Phe Lys Gln Cys Asp Thr Tyr Glu
690 695 700
Lys Leu Glu Lys Glu Val Asp Arg Lys Gly Tyr Lys Leu Glu Gly Cys
705 710 715 720
Lys Ser Glu Pro Lys Thr Ile Ser Leu Thr Gln Leu Thr Asp Trp Val
725 730 735
Glu Asn Lys Asp Cys Phe Leu Leu Pro Ile Val Asn Gln Asp Ile Asn
740 745 750
Lys Gly Asp Lys Arg Thr Lys Asn Gln Asn Gln Phe Thr Lys Asp Trp
755 760 765
Phe Asp Ile Phe Glu Asn Lys Lys Arg Leu His Pro Glu Phe Asn Ile
770 775 780
Phe Tyr Arg Phe Pro Thr Lys Asp Tyr Pro Asn Thr Lys Phe Lys Asn
785 790 795 800
Gly Thr Glu Lys Thr Lys Arg Tyr Ser Arg Phe Gln Met Leu Ala Tyr
805 810 815
Phe Gly Cys Glu Val Ile Pro Ser Gly Asn His Leu Ser Lys Lys Glu
820 825 830
Gln Ile Ala Ile Phe Asn Asn Asp Lys Lys Gln Lys Glu Glu Val Glu
835 840 845
Lys Tyr Asn Lys Ser Ile Ser Ser Asp Cys Asp Tyr Val Ile Gly Ile
850 855 860
Asp Arg Gly Ile Lys Gln Leu Ala Thr Leu Cys Val Leu Asp Lys Asn
865 870 875 880
Gly Val Ile Gln Gly Asp Phe Gln Ile Phe Thr Arg Thr Phe Asn Lys
885 890 895
Gln Thr Lys Gln Trp Glu His Lys Glu Leu Glu Gln Arg Asn Ile Leu
900 905 910
Asp Leu Ser Asn Leu Arg Val Glu Thr Thr Ile Thr Gly Lys Lys Val
915 920 925
Leu Val Asp Leu Ser Lys Ile Lys Asp Asp Glu Gly Asn Tyr Thr Asn
930 935 940
Leu Lys Gln Thr Ile Lys Leu Lys Gln Leu Ala Tyr Ile Arg Glu Leu
945 950 955 960
Gln Tyr Ala Met Gln Thr Arg Pro Asp Asp Leu Leu Asp Phe Val Lys
965 970 975
Ser Ile Asn Ser Ala Asn Asp Ile Thr Ala Glu Asn Ile Lys His Phe
980 985 990
Ile Ser Pro Tyr Lys Glu Gly Lys Asn Tyr Asp Asp Leu Pro Lys Val
995 1000 1005
Glu Met Phe Asn Leu Leu Lys Glu Trp Gly Asn Ala Asp Glu Asn
1010 1015 1020
Gly Lys Arg Lys Ile Ala Glu Leu Asp Pro Ala Asp Asn Leu Lys
1025 1030 1035
Ser Gly Ile Val Ala Asn Met Val Gly Val Val Ala Phe Leu Cys
1040 1045 1050
Glu Asn Tyr Asn Tyr Lys Val Arg Ile Ala Leu Glu Asp Leu Thr
1055 1060 1065
Arg Ala Tyr Gly Ile Gln Lys Asp Ala Leu Asn Gly Thr Ala Ile
1070 1075 1080
Tyr Gln Asn Asp Glu Asp Phe Lys Glu Gln Glu Asn Arg Arg Leu
1085 1090 1095
Ala Gly Val Gly Thr Met Gln Phe Phe Glu Val Gln Leu Leu Arg
1100 1105 1110
Lys Leu Phe Lys Ile Gln Val Asp Lys Asn Leu His Leu Ile Pro
1115 1120 1125
Ala Phe Arg Ser Val Asp Asn Tyr Glu Lys Ile Val Arg Arg Asp
1130 1135 1140
Lys Gln Asn Ser Gly Asp Glu Phe Val Asn Tyr Pro Phe Gly Ile
1145 1150 1155
Val Cys Phe Val Asp Pro Lys Tyr Thr Ser Gln Gln Cys Pro Tyr
1160 1165 1170
Cys Asn Asn Thr His Lys His Lys Lys Asn Asp Thr Glu Thr Gly
1175 1180 1185
Lys Lys Ala Phe Tyr Arg Asn Lys Gly Glu Asn Lys Asn Ser Leu
1190 1195 1200
Leu Cys Glu Lys Cys Gly Val Ser Thr Ile Glu Gly Glu Glu Thr
1205 1210 1215
Leu Ser Ser Lys Asn Asp Asn Lys Lys Gln Phe Asn Ile His Tyr
1220 1225 1230
Ile Thr Asp Gly Asp Gln Asn Gly Ala Tyr His Ile Ala Asn Lys
1235 1240 1245
Val Val Ile Asn Phe Gln Lys Asp Ser
1250 1255
<210> 7
<211> 1011
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.1的氨基酸序列
<400> 7
Met Asp Tyr Gln Gln Tyr Glu Phe Thr Arg Thr Ile Arg Phe Asn Leu
1 5 10 15
Ser Gly Asp Asp Lys Arg Ala Leu Met Leu Asp Leu Leu Asp Asp Thr
20 25 30
Gln Glu Gly Met Leu Ala Ala Phe Gln Glu Thr Tyr Lys Asn Leu Leu
35 40 45
Phe Ala Phe Gln Glu Ala Ile Leu Arg Ala Asp Gly Ser Gly Asn Leu
50 55 60
Arg Val Gly Arg Leu Glu Ile Lys Lys Ser Trp Leu Arg Gln Tyr Ala
65 70 75 80
Arg Glu Tyr Phe Tyr Ala Leu Ser Glu Asp Glu Arg Arg Cys Lys Asn
85 90 95
Lys Phe Gln Ala Lys Leu Phe Asp Arg Val Leu Ser Asp Trp Leu Glu
100 105 110
Arg Asn Asn Glu Leu Leu Gln Arg Leu Asn Asn Ile Leu Ser Leu Pro
115 120 125
Gln Glu Ser Lys Thr Gly Ala Ser Asp Leu Ser Leu Leu Val Arg Gln
130 135 140
Leu Lys Gly Ala Glu Tyr Phe Tyr Phe Ile Arg Asp Phe Thr Gln Ser
145 150 155 160
Gly Ile Ile Asn Asp Lys Asp Ser Asp Glu His Ile Lys Asn Leu Ala
165 170 175
Gly Ile Val Glu Lys Phe Glu Thr Leu Leu Asp Lys Val Leu Phe Leu
180 185 190
Thr Ala Pro Asn Ser Ser Gln Gly Val Glu Thr Thr Arg Ala Ser Phe
195 200 205
Asn Tyr Tyr Thr Val Asn Lys Ile Ser Lys Asn Phe Asp Glu Asn Ile
210 215 220
Lys Lys Ala Asn Gly Arg Leu Cys Ser Ser Tyr Gln Asn Ser Met Asn
225 230 235 240
Glu Glu Leu Leu Arg Lys Val Gly Phe Leu Lys Tyr Leu Lys Asp Glu
245 250 255
Tyr Arg Ala Glu Leu Gln Asn Val Ser Leu Lys Asp Leu Tyr Glu Ala
260 265 270
Leu Lys Lys Phe Lys Ser Gln Gln Lys Thr Ala Phe Ile Gln Ala Val
275 280 285
Gln Lys Asn Lys Ser Glu Lys Glu Leu Met Arg Glu Phe Pro Leu Phe
290 295 300
Asn Gly Lys Gln Pro Asp Thr Leu Gln Lys Phe Ile Leu Glu Thr Asp
305 310 315 320
Lys Ile Lys Arg Gly Ala Tyr Phe Gln Lys Trp Gly Phe Asp Asn Tyr
325 330 335
Ile Ser Phe Cys Asn Lys Ile Phe Lys Pro Val Ala Met Glu Thr Gly
340 345 350
Thr Arg Lys Ala Lys Ile Arg Ala Leu Glu Gln Glu Lys Ile Glu Ala
355 360 365
Arg Leu Leu Gln Tyr Trp Ala His Ile Leu Val Lys Asp Gly Lys Tyr
370 375 380
Phe Leu Leu Leu Ile Pro Lys Glu Lys Met Gly Glu Ala Lys Val Phe
385 390 395 400
Phe Ala Arg Leu Ser Asp Gln Glu Gly Gly Glu Tyr Thr Leu Tyr Ala
405 410 415
Phe Asn Ser Leu Thr Leu Arg Ala Leu Lys Lys Leu Ile Arg Arg Asn
420 425 430
Leu Gly Lys Glu Gln Val Arg Leu Ser Ala Gly Asp Ala Asp Ala Ile
435 440 445
Ala Leu Cys Gln Glu Val Leu Arg Gly Arg Tyr His Gln Leu Lys Asp
450 455 460
Leu Asp Leu Ser Gly Phe Glu Lys Glu Ile Ala Glu Ile Ala Asn Thr
465 470 475 480
Gln Tyr Glu Asn Glu Glu Glu Phe Arg Ile Ala Leu Glu Gln Val Ala
485 490 495
Tyr Tyr Leu Ser Glu Arg Lys Met Asn Glu Glu Ser Ile Glu Tyr Leu
500 505 510
Lys Lys Asn Leu Gly Ala Ile Leu Leu Glu Ile Ser Ser Tyr Asp Leu
515 520 525
Glu Arg Asn Ile Thr Gly Glu Ser Lys Glu His Thr Arg Leu Trp Ser
530 535 540
Asp Phe Trp Asn Pro Asn Asn Lys Lys Glu Cys Phe Ser Thr Arg Leu
545 550 555 560
Asn Pro Glu Leu Arg Ile Phe Tyr Arg Pro Pro Arg Glu Gln Lys Asp
565 570 575
Pro Lys Lys Gln Lys Asn Arg Phe Ser Lys Asp His Leu Ala Val Ala
580 585 590
Phe Thr Ile Ala Gln Asn Ala Ala Arg Lys Arg Met Glu Thr Ser Phe
595 600 605
Ala Glu Glu Lys Asp Leu Val Glu Gln Val Lys Lys Phe Asn Glu Glu
610 615 620
Val Val Gly Lys Phe Ile Asp Glu Lys Ser Asp Asn Leu Tyr Tyr Tyr
625 630 635 640
Gly Ile Asp Arg Gly Gln Gln Glu Leu Ala Thr Leu Cys Val Val Arg
645 650 655
Phe Ser Lys Glu His Tyr Glu Ala Met Leu Glu Asp Asn Phe Ile Lys
660 665 670
Lys Phe Ser Lys Pro Ile Pro Ala Gln Ile Thr Ala Tyr Arg Ile Lys
675 680 685
Asp Glu His Met Ser Tyr Arg Lys Asn Ile Thr Arg Asp Leu Lys Gly
690 695 700
Asn Glu Thr Glu Glu Ile Leu Phe Lys Asn Pro Ser His Phe Ile Asp
705 710 715 720
Glu Val Glu Asn Phe Glu Glu Val Ser Thr Pro Cys Ile Asp Leu Thr
725 730 735
Thr Ala Lys Leu Ile Lys Gly Lys Ile Ile Leu Asn Gly Asp Ile Gln
740 745 750
Thr Tyr Leu Ala Leu Lys Lys Ala Asn Gly Lys Arg Gln Leu Phe Glu
755 760 765
Lys Phe Ala Lys Ile Asp Asp Ser Ala Lys Ile Glu Phe Asp Asp Ser
770 775 780
Glu Gly Arg Phe Gln Val Lys Ser Lys Ala Thr Glu Arg Glu Glu Tyr
785 790 795 800
Gln Phe Leu Pro Tyr Tyr Gly Pro Glu Gln Glu Asn Ile Ser Pro Arg
805 810 815
Glu Asp Met Arg Arg Glu Leu Gln Ala Tyr Leu Asp Lys Leu Arg Ser
820 825 830
Ser Glu Ser Phe Glu Glu Asp Ile Ser Ile Glu Lys Ile Asn His Leu
835 840 845
Arg Asp Ala Ile Thr Ser Asn Met Val Gly Ile Ile Ala Phe Leu Phe
850 855 860
Thr Glu Tyr Pro Gly Ile Ile Asn Leu Glu Asn Leu His Ser Arg Glu
865 870 875 880
Asn Ile Glu Lys Asn Trp Arg Lys Asn Asn Glu Asp Ile Ser Arg Arg
885 890 895
Leu Glu Trp Gly Leu Tyr Lys Lys Phe Gln Lys Ile Gly Leu Val Pro
900 905 910
Pro Arg Leu Arg Gln Thr Val Leu Leu Arg Glu Asn Glu Thr Glu Arg
915 920 925
Gln Glu Lys Leu Asn Gln Phe Gly Ile Ile His Phe Ile Pro Thr Glu
930 935 940
Lys Thr Ser Ala Arg Cys Pro Tyr Cys Gly Glu Asn Thr Pro Met Lys
945 950 955 960
Gln Arg Asn Glu Asp Lys Phe Lys Leu His Ala Tyr Ile Cys Arg Ser
965 970 975
Asn Glu Glu Asn Cys Gly Phe Asp Thr Arg Glu Pro Lys Ser Pro Leu
980 985 990
Glu Phe Ile Lys Asn Ser Asp Asp Val Ala Ala Tyr Asn Ile Ala Lys
995 1000 1005
Lys Arg Leu
1010
<210> 8
<211> 1067
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.2的氨基酸序列
<400> 8
Met Lys Asn Gly Ile Asn Leu Phe Lys Thr Lys Thr Thr Lys Thr Lys
1 5 10 15
Gly Val Asp Met Glu Lys Tyr Gln Ile Thr Lys Thr Ile Arg Phe Lys
20 25 30
Leu Leu Pro Asp Asn Ala His Glu Ile Val Glu Lys Val Lys Ser Leu
35 40 45
Lys Thr Ser Asn Val Asp Glu Leu Met Asp Glu Val Lys Asn Val His
50 55 60
Leu Lys Gly Leu Glu Leu Leu Phe Ala Leu Lys Lys Tyr Phe Tyr Phe
65 70 75 80
Asp Gly Asn Gln Cys Lys Ser Phe Lys Ser Thr Leu Glu Ile Lys Ala
85 90 95
Arg Trp Leu Arg Leu Tyr Thr Pro Asp Gln Tyr Tyr Leu Lys Lys Ser
100 105 110
Ser Lys Asn Ser Tyr Gln Leu Lys Ser Leu Ser Tyr Phe Lys Asp Val
115 120 125
Phe Asn Asp Trp Leu Phe Asn Trp Glu Glu Ser Val Ser Glu Leu Ala
130 135 140
Ile Ile Tyr Glu Lys Tyr Lys Ile Cys Gln His Gln Arg Asp Ser Arg
145 150 155 160
Ala Asp Ile Ala Leu Leu Ile Lys Lys Leu Ser Met Lys Glu Tyr Phe
165 170 175
Pro Phe Ile Ser Asp Leu Ile Asp Cys Val Asn Asp Lys Asn Ser Asn
180 185 190
Lys Thr Phe Leu Met Lys Leu Ser Glu Glu Leu Ser Val Leu Leu Glu
195 200 205
Lys Cys Asn Ser Arg Ala Leu Pro Tyr Gln Ser Asn Gly Ile Val Val
210 215 220
Gly Lys Ala Ser Leu Asn Tyr Tyr Thr Val Ser Lys Ser Glu Lys Met
225 230 235 240
Leu Gln Asn Glu Tyr Glu Asp Val Cys Gln Ser Leu Asp Lys Asn Tyr
245 250 255
Asp Ile Thr Glu Met Lys Val Ile Leu Tyr Lys Glu Lys Leu Asp Asn
260 265 270
Leu Asn Phe Lys Asp Val Thr Ile Ala Asn Ala Tyr Asn Leu Leu Lys
275 280 285
Glu Asn Lys Ala Leu Gln Lys Arg Leu Phe Ser Glu Tyr Val Ser Gln
290 295 300
Gly Lys Val Leu Ser Leu Ile Lys Thr Glu Leu Pro Leu Phe Ser Asn
305 310 315 320
Ile Asn Asp Asn Asp Phe Glu Lys Tyr Lys Glu Trp Ser Asn Glu Ile
325 330 335
Lys Lys Leu Ala Asp Lys Lys Asn Thr Phe Cys Lys Lys Thr Gln Gln
340 345 350
Asp Lys Ile Lys Asp Ile Gln Asn Lys Ile Ser Glu Leu Lys Lys Lys
355 360 365
Arg Gly Ala Leu Phe Gln Tyr Lys Phe Thr Ser Phe Gln Lys His Cys
370 375 380
Asp Asn Tyr Lys Lys Val Ala Val Gln Tyr Gly Lys Leu Lys Ala Arg
385 390 395 400
Lys Lys Ala Ile Glu Lys Asp Glu Ile Glu Ala Asn Leu Leu Arg Tyr
405 410 415
Trp Ser Val Ile Leu Glu Gln Glu Asp Lys His Ser Leu Val Leu Ile
420 425 430
Pro Lys Asn Asn Ala Lys Asp Ala Lys Gln Tyr Ile Glu Thr Ile Asn
435 440 445
Thr Lys Gly Gly Lys Tyr Ile Ile His His Leu Asp Ser Leu Thr Leu
450 455 460
Arg Ala Leu Asn Lys Leu Cys Phe Asn Ala Val Asp Ile Glu Lys Gly
465 470 475 480
Gln Met Val Arg Glu Asn Thr Phe Tyr Gln Gly Ile Lys Glu Glu Phe
485 490 495
Glu Arg Asn Lys Ile Asn Cys Asp Asn Gln Gly Val Leu Lys Ile Gln
500 505 510
Gly Leu Tyr Ser Phe Lys Thr Glu Gly Gly Gln Ile Asn Glu Lys Glu
515 520 525
Ala Val Glu Phe Phe Lys Glu Val Leu Lys Ser Asn Tyr Ala Arg Glu
530 535 540
Val Leu Asn Leu Pro Tyr Asp Leu Glu Ser Asn Ile Phe Gln Lys Glu
545 550 555 560
Tyr Thr Asn Leu Asp Gln Phe Arg Gln Asp Leu Glu Lys Cys Cys Tyr
565 570 575
Ala Leu His Ser Lys Ile Gly Lys Asp Asp Leu Asp Glu Phe Thr Arg
580 585 590
Arg Phe Glu Ala Gln Val Phe Asp Ile Thr Ser Ile Asp Leu Lys Ser
595 600 605
Lys Lys Glu Lys Thr Lys Thr Thr Gly Glu Met Lys Lys His Thr Gln
610 615 620
Leu Trp Leu Glu Phe Trp Lys Gly Ala Ile Glu Gln Asn Phe Ala Thr
625 630 635 640
Arg Val Asn Pro Glu Leu Ser Ile Phe Trp Arg Ala Pro Lys Ser Ser
645 650 655
Arg Glu Lys Lys Tyr Gly Lys Gly Ser Asp Leu Tyr Asp Pro Asn Lys
660 665 670
Asn Asn Arg Tyr Leu Tyr Glu Gln Tyr Thr Leu Ala Leu Thr Ile Thr
675 680 685
Glu Asn Ala Gly Ser His Phe Lys Asp Ile Ala Phe Lys Asp Thr Ser
690 695 700
Lys Ile Lys Glu Ala Ile Lys Glu Phe Asn Met Ser Leu Ser Gln Ser
705 710 715 720
Lys Tyr Cys Phe Gly Ile Asp Arg Gly Asn Ala Glu Leu Val Ser Leu
725 730 735
Cys Leu Ile Lys Asn Glu Lys Asp Phe Pro Phe Glu Lys Phe Pro Val
740 745 750
Tyr Arg Leu Arg Asp Leu Thr Tyr Gln Gly Asp Phe Lys Asp Lys His
755 760 765
Asp Gln Met Arg Tyr Gly Val Ala Ile Lys Asn Ile Ser Tyr Phe Ile
770 775 780
Asp Gln Glu Asp Leu Phe Glu Lys Asn Asn Leu Ser Ala Ile Asp Met
785 790 795 800
Thr Thr Ala Lys Leu Ile Lys Asn Lys Ile Val Leu Asn Gly Asp Val
805 810 815
Leu Thr Tyr Leu Lys Leu Lys Glu Glu Thr Ala Lys His Lys Leu Thr
820 825 830
Gln Phe Phe Gln Gly Ser Ser Ile Asn Lys Asn Ser Arg Val Tyr Phe
835 840 845
Asp Glu Asp Glu Asn Val Phe Lys Ile Thr Thr Asn Arg Asn His Asn
850 855 860
Pro Glu Glu Ile Ile Tyr Phe Tyr Arg Gly Glu Tyr Gly Ala Ile Lys
865 870 875 880
Asn Lys Asn Asp Leu Glu Asp Ile Leu Asn Glu Tyr Leu Cys Lys Met
885 890 895
Glu Thr Gly Glu Ser Glu Ile Val Leu Leu Asn Arg Val Asn His Leu
900 905 910
Arg Asp Ala Ile Ser Ala Asn Ile Val Gly Ile Leu Ser Tyr Leu Ile
915 920 925
Asp Leu Phe Pro Glu Thr Ile Val Ala Leu Glu Asn Leu Ala Lys Gly
930 935 940
Thr Ile Asp Arg His Val Ser Gln Ser Tyr Glu Asn Ile Thr Arg Arg
945 950 955 960
Phe Glu Trp Ala Leu Tyr Arg Lys Leu Leu Asn Lys Gln Leu Ala Pro
965 970 975
Pro Glu Leu Lys Glu Asn Ile Leu Leu Arg Glu Gly Asp Asp Lys Ile
980 985 990
Asp Gln Phe Gly Ile Ile His Phe Val Glu Glu Lys Asn Thr Ser Lys
995 1000 1005
Asp Cys Pro Asn Cys Arg Lys Thr Thr Gln Gln Thr Asn Asp Asn
1010 1015 1020
Lys Phe Lys Glu Lys Lys Phe Val Cys Lys Ser Cys Gly Phe Asp
1025 1030 1035
Thr Ser Lys Asp Arg Lys Gly Met Asp Ser Leu Asn Ser Pro Asp
1040 1045 1050
Thr Val Ala Ala Tyr Asn Val Ala Arg Lys Lys Phe Glu Ser
1055 1060 1065
<210> 9
<211> 1091
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.3的氨基酸序列
<400> 9
Met Ala Gly Thr Pro Tyr Thr Gly His Val Ala Cys Lys Tyr Cys Lys
1 5 10 15
Ile Thr Ser Trp Ala Thr Tyr Asp Arg Ile Lys Ile Asn Lys Ile Asn
20 25 30
Met Asn Gln Ser Phe Ile Asn Gly Gln Asn Phe Tyr Glu Leu Arg Lys
35 40 45
Thr Ile Arg Phe Val Leu Asp Pro Lys Thr Leu Lys Arg Pro Tyr Thr
50 55 60
Pro Ser Ser Asp Glu Val Asn Leu Glu Glu Gln Leu Asn Asn Phe Ile
65 70 75 80
Glu Lys Tyr Gln Gln Gly Ile Asn Asp Phe Lys Tyr Ile Val Tyr Phe
85 90 95
Gly Pro Lys Thr Ala Glu Thr Lys Glu Leu Asn Lys Lys Ile Ser Ile
100 105 110
Lys His Ser Trp Leu Arg Asn Tyr Thr Lys Ser Glu Phe Tyr Ser Ile
115 120 125
Lys Asp Lys Leu Ile Gln Leu Asp Tyr Asn Gly Asn Lys Ala Ser Ile
130 135 140
Gly Asn Ser Asn Leu Lys Phe Leu Asn Glu Tyr Phe Glu Asn Trp Ile
145 150 155 160
Ser Glu Asn Gln Glu Cys Ala Asp Ala Leu Lys Asn Cys Ile Asn Ala
165 170 175
Pro Ala Glu Lys Gln Lys Arg Lys Ser Glu Ala Ala His Trp Val Arg
180 185 190
Lys Leu Thr Lys Arg Ser Asn Phe Glu Cys Ile Phe Glu Leu Phe Asn
195 200 205
Gly Asn Ile Asp His Lys Asn Ser Asn Asp Asp Ile Glu Lys Ile Lys
210 215 220
His Cys Leu Asn Glu Cys Lys Thr Leu Leu Thr Ser Leu Glu Lys Met
225 230 235 240
Leu Leu Pro Ser Gln Ser Leu Gly Met Glu Ile Glu Arg Ala Ser Leu
245 250 255
Asn Tyr Tyr Thr Ile Asn Lys Lys Pro Lys Asn Tyr Asp Glu Asp Ile
260 265 270
Ala Gln Lys Ala Ser Ala Leu Asn Glu Ala Tyr Gln Phe Lys Ala Asp
275 280 285
Asp Lys Ala Phe Leu Asn Arg Val Gly Phe Ser Asp Asp Gly Val Pro
290 295 300
Ile Asn Glu Leu Lys Glu Ala Met Lys Lys Phe Lys Ala Asp Gln Lys
305 310 315 320
Ser Lys Phe Tyr Glu Phe Val Asn Gln Lys Lys Ser Tyr Ser Asp Leu
325 330 335
Lys Lys Asn Asp Asp Leu Lys Leu Leu Asn Asp Ile Ser Glu Glu Asp
340 345 350
Phe Asn Lys Phe Lys Glu Thr Gln Asp Lys Met Thr Arg Gly Lys His
355 360 365
Phe Gln Phe Ser Phe Pro Asn Tyr Lys Lys Ser Glu Lys Asn Phe Cys
370 375 380
Asp Leu Tyr Lys Asn Val Ala Val Ala Phe Gly Lys Ile Arg Ala Asp
385 390 395 400
Ile Lys Ala Leu Glu Lys Glu Arg Met Asp Ala Glu Lys Leu Gln Cys
405 410 415
Trp Ala Val Ile Leu Glu Lys Asp Asn Gln Arg Tyr Val Val Thr Ile
420 425 430
Pro Arg Asp Ala Asn Asn Asn Leu Thr Asn Thr Lys Gln Tyr Ile Asp
435 440 445
Asn Leu Gln Asn Glu Glu Asn Asp Gln Trp Ile Leu Tyr Ala Phe Glu
450 455 460
Ser Leu Thr Leu Arg Ser Leu Asp Lys Leu Cys Phe Gly Leu Asp Lys
465 470 475 480
Asn Thr Phe Ile Pro Ala Ile Thr Gly Glu Leu Tyr Gln Lys Asn Asn
485 490 495
Ser Phe Phe Glu Lys Gly Leu Leu Lys Arg Lys Asp Gln Phe Ser Gln
500 505 510
Asn Gly Thr Asp Leu Ala Ala Phe Tyr Lys Thr Val Leu Glu Leu Asp
515 520 525
Ser Thr Lys Lys Met Leu Gly Ile Asn Lys Tyr Ala Asp Phe Lys Ala
530 535 540
Phe Ile Ser Lys Glu Tyr Thr Ala Leu Glu Asp Phe Glu Lys Thr Leu
545 550 555 560
Lys Glu Thr Cys Tyr Phe Lys Lys Arg Val Phe Ile Ser Glu Asp Thr
565 570 575
Lys Asn Lys Leu Ile Asn Asp Tyr Gln Gly Asn Leu Tyr Lys Ile Thr
580 585 590
Ser Tyr Asp Leu Glu Lys Asp Asp Ser Glu Ala Leu Gly Thr Leu Ile
595 600 605
Asn Lys Lys Gln Phe Asn Arg Ala Ser Pro Glu Ile His Thr Lys Thr
610 615 620
Trp Leu Asp Phe Trp Thr Ala Asp Asn Glu Thr Asp Lys Tyr Pro Ile
625 630 635 640
Arg Leu Asn Pro Glu Phe Lys Ile Ser Phe Val Glu Lys Gln Asp Lys
645 650 655
Asp Leu Asn Met Arg Asn Leu Gly Leu Leu Asn Lys Asn Arg Arg Leu
660 665 670
Lys Ser Gln Phe Leu Leu Ser Thr Thr Ile Thr Leu Leu Ala His Glu
675 680 685
Lys Asn Ala Asp Leu His Phe Lys Lys Thr Asp Glu Ile Gln Thr Phe
690 695 700
Ile Asn Ser Tyr Asn Gln Glu Phe Asn Lys Lys Ile Lys Pro Phe Asp
705 710 715 720
Ile Tyr Tyr Tyr Gly Leu Asp Arg Gly Gln Lys Glu Leu Leu Thr Leu
725 730 735
Gly Leu Phe Lys Phe Ser Glu Asn Glu Lys Val Ser Phe Thr Lys Gln
740 745 750
Asp Gly Thr Val Gly Glu Tyr Ser Lys Pro Lys Phe Ile Pro Leu Asp
755 760 765
Val Tyr Gln Ile Arg Glu Gly Gln Tyr Leu Thr Lys Asn Lys Lys Gly
770 775 780
Arg Leu Ala Tyr Lys Ser Ile Asp Gln Phe Ile Asp Asp Glu Lys Val
785 790 795 800
Ile Glu Lys Leu Pro Val Asn Ser Cys Leu Asp Leu Ser Cys Ala Lys
805 810 815
Leu Val Lys Gly Lys Ile Ile Gln Asn Gly Asp Val Ala Thr Tyr Leu
820 825 830
Glu Leu Lys Arg Val Ser Ala Leu Arg Lys Ile Tyr Glu Asn Thr Thr
835 840 845
Arg Gly Gln Phe Lys Thr Asp Arg Ile Gly Phe Asn Lys Asp Lys Gly
850 855 860
Cys Leu Phe Leu Asp Ile Glu Asn Arg Gly Lys Leu Glu Asn Asn Asn
865 870 875 880
Leu Tyr Phe Tyr Asp Asn Arg Phe Ala Glu Ile Leu Ser Leu Asp Ser
885 890 895
Ile Ile Lys Glu Leu Gln Asp Tyr Tyr Asn Glu Val Lys Asn Lys Gln
900 905 910
Asn Ile Glu Phe Ile Ser Ile Asp Lys Ile Asn His Leu Arg Asp Ala
915 920 925
Leu Cys Ala Asn Ala Val Gly Ile Leu Ala His Leu Gln Lys Thr His
930 935 940
Phe Gly Val Ile Val Phe Glu Gly Leu Asp Ala Arg His Lys Asn Lys
945 950 955 960
Glu Thr Thr Glu Phe Ala Gly Asn Leu Ala Ser Arg Ile Glu Arg Lys
965 970 975
Ile Leu Gln Lys Leu Glu Thr Leu Ser Leu Ile Pro Pro Gln His Arg
980 985 990
Gln Ile Ile Asp Leu Gln Asn Ser Lys Gln Ile Lys Gln Thr Gly Ala
995 1000 1005
Val Leu Tyr Ile Glu Glu Lys Gly Thr Ser Ala Asn Cys Pro His
1010 1015 1020
Cys Glu Thr Ala Asn Pro Asp Lys Ser Glu Lys Trp Leu Ala His
1025 1030 1035
Asn Tyr Lys Cys Lys Asn Ser Asn Cys Asn Phe Asp Ala Ser Glu
1040 1045 1050
Ile Ser Lys Arg Lys Asp Leu Ile Gly Leu Asp Asn Ser Asp Ser
1055 1060 1065
Val Ala Thr Tyr Asn Ile Ala Lys Arg Gly Leu Leu Glu Met Asn
1070 1075 1080
Gln Lys Ile Glu Gln Ser Lys Val
1085 1090
<210> 10
<211> 1057
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.4的氨基酸序列
<400> 10
Met Glu Ile Gln Glu Leu Lys Asn Leu Tyr Glu Val Lys Lys Thr Val
1 5 10 15
Arg Phe Glu Leu Lys Pro Ser Lys Lys Lys Ile Phe Glu Gly Gly Asp
20 25 30
Val Ile Lys Leu Gln Lys Asp Phe Glu Lys Val Gln Lys Phe Phe Leu
35 40 45
Asp Ile Phe Val Tyr Lys Asn Glu His Thr Lys Leu Glu Phe Lys Lys
50 55 60
Lys Arg Glu Ile Lys Tyr Thr Trp Leu Arg Thr Asn Thr Lys Asn Glu
65 70 75 80
Phe Tyr Asn Trp Arg Gly Lys Ser Asp Thr Gly Lys Asn Tyr Ala Leu
85 90 95
Asn Lys Ile Gly Phe Leu Ala Glu Glu Ile Leu Arg Trp Leu Asn Glu
100 105 110
Trp Gln Glu Leu Thr Lys Ser Leu Lys Asp Leu Thr Gln Arg Glu Glu
115 120 125
His Lys Gln Glu Arg Lys Ser Asp Ile Ala Phe Val Leu Arg Asn Phe
130 135 140
Leu Lys Arg Gln Asn Leu Pro Phe Ile Lys Asp Phe Phe Asn Ala Val
145 150 155 160
Ile Asp Ile Gln Gly Lys Gln Gly Lys Glu Ser Asp Asp Lys Ile Arg
165 170 175
Lys Phe Arg Glu Glu Ile Lys Glu Ile Glu Lys Asn Leu Asn Ala Cys
180 185 190
Ser Arg Glu Tyr Leu Pro Thr Gln Ser Asn Gly Val Leu Leu Tyr Lys
195 200 205
Ala Ser Phe Ser Tyr Tyr Thr Leu Asn Lys Thr Pro Lys Glu Tyr Glu
210 215 220
Asp Leu Lys Lys Glu Lys Glu Ser Glu Leu Ser Ser Val Leu Leu Lys
225 230 235 240
Glu Ile Tyr Arg Arg Lys Arg Phe Asn Arg Thr Thr Asn Gln Lys Asp
245 250 255
Thr Leu Phe Glu Cys Thr Ser Asp Trp Leu Val Lys Ile Lys Leu Gly
260 265 270
Lys Asp Ile Tyr Glu Trp Thr Leu Asp Glu Ala Tyr Gln Lys Met Lys
275 280 285
Ile Trp Lys Ala Asn Gln Lys Ser Asn Phe Ile Glu Ala Val Ala Gly
290 295 300
Asp Lys Leu Thr His Gln Asn Phe Arg Lys Gln Phe Pro Leu Phe Asp
305 310 315 320
Ala Ser Asp Glu Asp Phe Glu Thr Phe Tyr Arg Leu Thr Lys Ala Leu
325 330 335
Asp Lys Asn Pro Glu Asn Ala Lys Lys Ile Ala Gln Lys Arg Gly Lys
340 345 350
Phe Phe Asn Ala Pro Asn Glu Thr Val Gln Thr Lys Asn Tyr His Glu
355 360 365
Leu Cys Glu Leu Tyr Lys Arg Ile Ala Val Lys Arg Gly Lys Ile Ile
370 375 380
Ala Glu Ile Lys Gly Ile Glu Asn Glu Glu Val Gln Ser Gln Leu Leu
385 390 395 400
Thr His Trp Ala Val Ile Ala Glu Glu Arg Asp Lys Lys Phe Ile Val
405 410 415
Leu Ile Pro Arg Lys Asn Gly Gly Lys Leu Glu Asn His Lys Asn Ala
420 425 430
His Ala Phe Leu Gln Glu Lys Asp Arg Lys Glu Pro Asn Asp Ile Lys
435 440 445
Val Tyr His Phe Lys Ser Leu Thr Leu Arg Ser Leu Glu Lys Leu Cys
450 455 460
Phe Lys Glu Ala Lys Asn Thr Phe Ala Pro Glu Ile Lys Lys Glu Thr
465 470 475 480
Asn Pro Lys Ile Trp Phe Pro Thr Tyr Lys Gln Glu Trp Asn Ser Thr
485 490 495
Pro Glu Arg Leu Ile Lys Phe Tyr Lys Gln Val Leu Gln Ser Asn Tyr
500 505 510
Ala Gln Thr Tyr Leu Asp Leu Val Asp Phe Gly Asn Leu Asn Thr Phe
515 520 525
Leu Glu Thr His Phe Thr Thr Leu Glu Glu Phe Glu Ser Asp Leu Glu
530 535 540
Lys Thr Cys Tyr Thr Lys Val Pro Val Tyr Phe Ala Lys Lys Glu Leu
545 550 555 560
Glu Thr Phe Ala Asp Glu Phe Glu Ala Glu Val Phe Glu Ile Thr Thr
565 570 575
Arg Ser Ile Ser Thr Glu Ser Lys Arg Lys Glu Asn Ala His Ala Glu
580 585 590
Ile Trp Arg Asp Phe Trp Ser Arg Glu Asn Glu Glu Glu Asn His Ile
595 600 605
Thr Arg Leu Asn Pro Glu Val Ser Val Leu Tyr Arg Asp Glu Ile Lys
610 615 620
Glu Lys Ser Asn Thr Ser Arg Lys Asn Arg Lys Ser Asn Ala Asn Asn
625 630 635 640
Arg Phe Ser Asp Pro Arg Phe Thr Leu Ala Thr Thr Ile Thr Leu Asn
645 650 655
Ala Asp Lys Lys Lys Ser Asn Leu Ala Phe Lys Thr Val Glu Asp Ile
660 665 670
Asn Ile His Ile Asp Asn Phe Asn Lys Lys Phe Ser Lys Asn Phe Ser
675 680 685
Gly Glu Trp Val Tyr Gly Ile Asp Arg Gly Leu Lys Glu Leu Ala Thr
690 695 700
Leu Asn Val Val Lys Phe Ser Asp Val Lys Asn Val Phe Gly Val Ser
705 710 715 720
Gln Pro Lys Glu Phe Ala Lys Ile Pro Ile Tyr Lys Leu Arg Asp Glu
725 730 735
Lys Ala Ile Leu Lys Asp Glu Asn Gly Leu Ser Leu Lys Asn Ala Lys
740 745 750
Gly Glu Ala Arg Lys Val Ile Asp Asn Ile Ser Asp Val Leu Glu Glu
755 760 765
Gly Lys Glu Pro Asp Ser Thr Leu Phe Glu Lys Arg Glu Val Ser Ser
770 775 780
Ile Asp Leu Thr Arg Ala Lys Leu Ile Lys Gly His Ile Ile Ser Asn
785 790 795 800
Gly Asp Gln Lys Thr Tyr Leu Lys Leu Lys Glu Thr Ser Ala Lys Arg
805 810 815
Arg Ile Phe Glu Leu Phe Ser Thr Ala Lys Ile Asp Lys Ser Ser Gln
820 825 830
Phe His Val Arg Lys Thr Ile Glu Leu Ser Gly Thr Lys Ile Tyr Trp
835 840 845
Leu Cys Glu Trp Gln Arg Gln Asp Ser Trp Arg Thr Glu Lys Val Ser
850 855 860
Leu Arg Asn Thr Leu Lys Gly Tyr Leu Gln Asn Leu Asp Leu Lys Asn
865 870 875 880
Arg Phe Glu Asn Ile Glu Thr Ile Glu Lys Ile Asn His Leu Arg Asp
885 890 895
Ala Ile Thr Ala Asn Met Val Gly Ile Leu Ser His Leu Gln Asn Lys
900 905 910
Leu Glu Met Gln Gly Val Ile Ala Leu Glu Asn Leu Asp Thr Val Arg
915 920 925
Glu Gln Ser Asn Lys Lys Met Ile Asp Glu His Phe Glu Gln Ser Asn
930 935 940
Glu His Val Ser Arg Arg Leu Glu Trp Ala Leu Tyr Cys Lys Phe Ala
945 950 955 960
Asn Thr Gly Glu Val Pro Pro Gln Ile Lys Glu Ser Ile Phe Leu Arg
965 970 975
Asp Glu Phe Lys Val Cys Gln Ile Gly Ile Leu Asn Phe Ile Asp Val
980 985 990
Lys Gly Thr Ser Ser Asn Cys Pro Asn Cys Asp Gln Glu Ser Arg Lys
995 1000 1005
Thr Gly Ser His Phe Ile Cys Asn Phe Gln Asn Asn Cys Ile Phe
1010 1015 1020
Ser Ser Lys Glu Asn Arg Asn Leu Leu Glu Gln Asn Leu His Asn
1025 1030 1035
Ser Asp Asp Val Ala Ala Phe Asn Ile Ala Lys Arg Gly Leu Glu
1040 1045 1050
Ile Val Lys Val
1055
<210> 11
<211> 1158
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.5的氨基酸序列
<400> 11
Met Glu Asn Phe Lys Asn Leu Tyr Glu Val Arg Lys Thr Val Arg Phe
1 5 10 15
Glu Leu Lys Pro Ser Arg Lys Lys Thr Phe Ala Gly Gly Asp Ile Phe
20 25 30
Glu Leu Gln Lys Asp Phe Glu Glu Val Gln Lys Phe Phe Leu Asp Ile
35 40 45
Phe Val Phe Ala Ile Glu Gln Glu Lys Leu Tyr Gln Glu Glu Glu Glu
50 55 60
Glu Gly Lys Leu Ser Arg Tyr Thr Lys Ile Glu Phe Lys Lys Lys Arg
65 70 75 80
Glu Ile Lys Tyr Thr Trp Leu Arg Ile Tyr Thr Lys Asn Glu Phe Tyr
85 90 95
Asp Trp Asn Gly Lys Asn Asp Lys Glu Lys Asn Tyr Ala Leu Ser Lys
100 105 110
Ile Asp Phe Leu Glu Lys Glu Ile Leu Arg Trp Phe Asn Glu Trp Gln
115 120 125
Glu Leu Thr Val Asn Leu Lys Asn Leu Thr Gln Thr Lys Glu His Glu
130 135 140
Lys Glu Arg Lys Ser Asp Ile Ala Phe Val Leu Arg Asn Phe Leu Lys
145 150 155 160
Arg Gln Asn Phe Pro Phe Ile Lys Asp Phe Phe Asn Ala Val Ile Asp
165 170 175
Ile Gln Glu Lys Gln Gly Asn Glu Ser Asp Glu Lys Ile Arg Lys Phe
180 185 190
Arg Glu Glu Leu Arg Glu Met Lys Lys Asn Leu Asn Thr Cys Ala Lys
195 200 205
Glu Tyr Leu Ser Ser Gln Ser Lys Gly Val Leu Leu His Lys Ala Ser
210 215 220
Phe Asn Tyr Tyr Thr Leu Asn Lys Thr Pro Lys Glu Tyr Glu Asn Leu
225 230 235 240
Lys Leu Gln Lys Glu Leu Glu Ile Asp Asn Ile Leu Pro Lys Lys Ile
245 250 255
Cys Lys Arg Val Arg Trp Asn Lys Glu Lys Lys Gln Glu Asp Ile Leu
260 265 270
Phe Glu Cys Asn Ser Asp Trp Leu Val Glu Ile Lys Leu Gly Tyr Asp
275 280 285
Ile Gln Lys Trp Thr Leu Asp Glu Ala Tyr Gln Lys Met Lys Thr Trp
290 295 300
Lys Ala Asp Gln Lys Ser Asp Phe Asn Glu Lys Ile Gly Asn Phe Ile
305 310 315 320
Asp Gln Tyr Leu Lys Lys Gly Phe Ile Glu Asp Leu Met Asn Glu Asn
325 330 335
Glu Lys Lys Asn Ala Glu Ala Ile Leu Arg Glu Phe Ser Val Phe Lys
340 345 350
Pro Ile Glu Asn Phe Tyr Phe Tyr Asp Phe Leu Glu Arg Thr Lys Glu
355 360 365
Ile Lys Ile Leu Ser Asn Gln Lys Asn Asn Ile Leu Gln Lys Tyr Asn
370 375 380
Lys Asn Ala Lys Tyr Phe Glu Lys Ile Ile Thr Tyr Lys Ile Lys Asp
385 390 395 400
Lys Glu Asp Leu Thr Glu Asp Glu Lys Glu Tyr Gln Glu Leu Glu Lys
405 410 415
Ser Ile Glu Lys Lys Ala Lys Glu Arg Gly Lys Phe Phe Asn Ala Pro
420 425 430
Lys Glu Lys Val Gln Thr Gln His Tyr Phe Glu Leu Cys Glu Leu Tyr
435 440 445
Lys Arg Ile Ala Met Lys Arg Gly Lys Ile Ile Ala Glu Ile Lys Gly
450 455 460
Ile Glu Asn Glu Glu Val Gln Ser Gln Leu Leu Thr His Trp Ala Leu
465 470 475 480
Ile Ala Glu Glu Gly Glu Lys Lys Ser Val Val Phe Ile Pro Arg Lys
485 490 495
Asn Gly Glu Glu Leu Glu Asn His Lys Lys Ala His Glu Phe Leu Gln
500 505 510
Lys Gln Glu Lys Lys Glu Phe Gly Asp Ile Lys Ser Tyr His Phe Lys
515 520 525
Ser Leu Thr Leu Arg Ala Leu Glu Lys Leu Cys Phe Lys Glu Thr Glu
530 535 540
Asn Thr Phe Thr Pro Glu Ile Lys Lys Glu Thr Asn Pro Lys Val Trp
545 550 555 560
Phe Pro Lys Tyr Lys Gln Glu Trp Asn Asp Glu Pro Gln Lys Leu Ile
565 570 575
Asn Phe Tyr Lys Gln Val Leu Gln Ser Lys Tyr Ser Gln Lys Tyr Leu
580 585 590
Asp Leu Val Ala Phe Gly Asp Leu Lys Ser Phe Leu Glu Thr Ser Phe
595 600 605
Asp Asp Leu Gln Ile Phe Glu Ser Gly Leu Glu Lys Thr Cys Tyr Ile
610 615 620
Lys Val Pro Ile Tyr Phe Ser Lys Glu Gly Phe Glu Thr Phe Thr Asn
625 630 635 640
Arg Phe Asp Ala Glu Val Phe Glu Ile Thr Thr Arg Ser Ile Ser Ser
645 650 655
Glu Ser Lys Arg Lys Glu Asn Ala His Ala Glu Ile Trp Lys Asp Phe
660 665 670
Trp Ser Lys Glu Asn Glu Glu Lys Asn His Ile Thr Arg Leu Asn Pro
675 680 685
Glu Val Ser Val Phe Tyr Arg Asp Glu Ile Glu Lys Lys Ser Asn Ala
690 695 700
Leu Arg Gly Asn Asn Lys Ser Asn Ile Asn Asn Arg Phe Ser Ala Ser
705 710 715 720
Arg Phe Thr Leu Val Thr Thr Ile Thr Ile Arg Ala Thr His Lys Lys
725 730 735
Ser Asn Leu Ala Phe Lys Thr Glu Glu Asp Ile Lys Ser His Ile Asp
740 745 750
Lys Phe Asn Glu Ala Phe Gln Asn Phe Ser Gly Glu Trp Val Tyr Gly
755 760 765
Ile Asp Arg Gly Leu Lys Glu Leu Ala Thr Leu Asn Val Val Lys Phe
770 775 780
Ser Asp Glu Lys Asn Glu Phe Gly Val Ile Lys Pro Lys Glu Phe Ala
785 790 795 800
Lys Ile Pro Val Tyr Lys Leu Lys Asp Glu Lys Ala Ile Leu Lys Asp
805 810 815
Glu Asn Gly Lys Asp Leu Lys Asn Ala Lys Gly Glu Ala Arg Lys Val
820 825 830
Ile Asp Asn Ile Ser Glu Val Leu Glu Glu Lys Lys Glu Pro Asp Ser
835 840 845
Asn Leu Phe Glu Lys Gln Gly Val Leu Ser Gln Gly Ile Ser Cys Ile
850 855 860
Asp Leu Thr Gln Ala Lys Leu Ile Lys Gly His Ile Ile Leu Asn Gly
865 870 875 880
Asp Gln Lys Thr Tyr Leu Lys Leu Lys Glu Ile Ser Ala Lys Arg Arg
885 890 895
Ile Phe Glu Leu Phe Ser Thr Ser Lys Ile Asp Lys Asn Ser Glu Leu
900 905 910
Arg Val Glu Lys Thr Thr Ile Ser Ile Asn Ser Glu Asp Gly Lys Arg
915 920 925
Asp Phe Tyr Trp Leu Thr Lys Asn Gln Ile Val Asn Ser Glu Thr Lys
930 935 940
Lys Glu Ile Gln Lys Glu Gln Gln Glu Lys Leu Asp Asn Leu Lys Val
945 950 955 960
Ile Phe Ile Asp Tyr Leu Glu Gly Leu Cys Val Lys Asn Lys Phe Glu
965 970 975
Asp Ile Glu Thr Ile Glu Lys Ile Asn His Leu Arg Asp Ala Ile Thr
980 985 990
Ala Asn Met Val Gly Ile Leu Phe His Leu Gln Lys Glu Phe Lys Gly
995 1000 1005
Ile Ile Ala Leu Glu Asn Leu Asp Thr Val Arg Glu Gln Ser Asn
1010 1015 1020
Lys Lys Met Ile Asp Glu His Phe Glu Gln Ser Asn Glu Asp Ile
1025 1030 1035
Ser Arg Arg Leu Glu Trp Ala Leu Tyr Arg Lys Phe Ala Asn Met
1040 1045 1050
Gly Glu Val Pro Ser Gln Ile Lys Glu Ser Ile Phe Leu Arg Asp
1055 1060 1065
Glu Phe Lys Val Tyr Gln Met Gly Leu Leu Lys Phe Val Glu Val
1070 1075 1080
Ser Gly Thr Ser Ser Asn Cys Pro Asn Cys Asp Lys Glu Val Gly
1085 1090 1095
Lys Thr Asn Ser His Phe Val Cys Lys Gly Glu Asn Asn Cys Gly
1100 1105 1110
Phe Ser Ser Lys Glu Asn Arg Asn Leu Leu Glu Gln Asn Leu Asn
1115 1120 1125
Asn Ser Asp Glu Val Ala Ala Tyr Asn Ile Ala Lys Arg Gly Leu
1130 1135 1140
Lys Leu Ile Asn Gln Lys Trp Asn Asn Thr Ser Lys Ser Gln Asn
1145 1150 1155
<210> 12
<211> 1064
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.6的氨基酸序列
<400> 12
Met Glu Lys Tyr Lys Ile Thr Lys Thr Ile Arg Phe Lys Leu Leu Pro
1 5 10 15
Asp Lys Ile Gln Asp Ile Ser Arg Gln Val Ala Val Leu Gln Asn Ser
20 25 30
Thr Asn Ala Glu Lys Lys Asn Asn Leu Leu Arg Leu Val Gln Arg Gly
35 40 45
Gln Glu Leu Pro Lys Leu Leu Asn Glu Tyr Ile Arg Tyr Ser Asp Asn
50 55 60
His Lys Leu Lys Ser Asn Val Thr Val His Phe Arg Trp Leu Arg Leu
65 70 75 80
Phe Thr Lys Asp Leu Phe Tyr Asn Trp Lys Lys Asp Asn Thr Glu Lys
85 90 95
Lys Ile Lys Ile Ser Asp Val Val Tyr Leu Ser His Val Phe Glu Ala
100 105 110
Phe Leu Lys Glu Trp Glu Ser Thr Ile Glu Arg Val Asn Ala Asp Cys
115 120 125
Asn Lys Pro Glu Glu Ser Lys Thr Arg Asp Ala Glu Ile Ala Leu Ser
130 135 140
Ile Arg Lys Leu Gly Ile Lys His Gln Leu Pro Phe Ile Lys Gly Phe
145 150 155 160
Val Asp Asn Ser Asn Asp Lys Asn Ser Glu Asp Thr Lys Ser Lys Leu
165 170 175
Thr Ala Leu Leu Ser Glu Phe Glu Ala Val Leu Lys Ile Cys Glu Gln
180 185 190
Asn Tyr Leu Pro Ser Gln Ser Ser Gly Ile Ala Ile Ala Lys Ala Ser
195 200 205
Phe Asn Tyr Tyr Thr Ile Asn Lys Lys Gln Lys Asp Phe Glu Ala Glu
210 215 220
Ile Val Ala Leu Lys Lys Gln Leu His Ala Arg Tyr Gly Asn Lys Lys
225 230 235 240
Tyr Asp Gln Leu Leu Arg Glu Leu Asn Leu Ile Pro Leu Lys Glu Leu
245 250 255
Pro Leu Lys Glu Leu Pro Leu Ile Glu Phe Tyr Ser Glu Ile Lys Lys
260 265 270
Arg Lys Ser Thr Lys Lys Ser Glu Phe Leu Glu Ala Val Ser Asn Gly
275 280 285
Leu Val Phe Asp Asp Leu Lys Ser Lys Phe Pro Leu Phe Gln Thr Glu
290 295 300
Ser Asn Lys Tyr Asp Glu Tyr Leu Lys Leu Ser Asn Lys Ile Thr Gln
305 310 315 320
Lys Ser Thr Ala Lys Ser Leu Leu Ser Lys Asp Ser Pro Glu Ala Gln
325 330 335
Lys Leu Gln Thr Glu Ile Thr Lys Leu Lys Lys Asn Arg Gly Glu Tyr
340 345 350
Phe Lys Lys Ala Phe Gly Lys Tyr Val Gln Leu Cys Glu Leu Tyr Lys
355 360 365
Glu Ile Ala Gly Lys Arg Gly Lys Leu Lys Gly Gln Ile Lys Gly Ile
370 375 380
Glu Asn Glu Arg Ile Asp Ser Gln Arg Leu Gln Tyr Trp Ala Leu Val
385 390 395 400
Leu Glu Asp Asn Leu Lys His Ser Leu Ile Leu Ile Pro Lys Glu Lys
405 410 415
Thr Asn Glu Leu Tyr Arg Lys Val Trp Gly Ala Lys Asp Asp Gly Ala
420 425 430
Ser Ser Ser Ser Ser Ser Thr Leu Tyr Tyr Phe Glu Ser Met Thr Tyr
435 440 445
Arg Ala Leu Arg Lys Leu Cys Phe Gly Ile Asn Gly Asn Thr Phe Leu
450 455 460
Pro Glu Ile Gln Lys Glu Leu Pro Gln Tyr Asn Gln Lys Glu Phe Gly
465 470 475 480
Glu Phe Cys Phe His Lys Ser Asn Asp Asp Lys Glu Ile Asp Glu Pro
485 490 495
Lys Leu Ile Ser Phe Tyr Gln Ser Val Leu Lys Thr Asp Phe Val Lys
500 505 510
Asn Thr Leu Ala Leu Pro Gln Ser Val Phe Asn Glu Val Ala Ile Gln
515 520 525
Ser Phe Glu Thr Arg Gln Asp Phe Gln Ile Ala Leu Glu Lys Cys Cys
530 535 540
Tyr Ala Lys Lys Gln Ile Ile Ser Glu Ser Leu Lys Lys Glu Ile Leu
545 550 555 560
Glu Asn Tyr Asn Thr Gln Ile Phe Lys Ile Thr Ser Leu Asp Leu Gln
565 570 575
Arg Ser Glu Gln Lys Asn Leu Lys Gly His Thr Arg Ile Trp Asn Arg
580 585 590
Phe Trp Thr Lys Gln Asn Glu Glu Ile Asn Tyr Asn Leu Arg Leu Asn
595 600 605
Pro Glu Ile Ala Ile Val Trp Arg Lys Ala Lys Lys Thr Arg Ile Glu
610 615 620
Lys Tyr Gly Glu Arg Ser Val Leu Tyr Glu Pro Glu Lys Arg Asn Arg
625 630 635 640
Tyr Leu His Glu Gln Tyr Thr Leu Cys Thr Thr Val Thr Asp Asn Ala
645 650 655
Leu Asn Asn Glu Ile Thr Phe Ala Phe Glu Asp Thr Lys Lys Lys Gly
660 665 670
Thr Glu Ile Val Lys Tyr Asn Glu Lys Ile Asn Gln Thr Leu Lys Lys
675 680 685
Glu Phe Asn Lys Asn Gln Leu Trp Phe Tyr Gly Ile Asp Ala Gly Glu
690 695 700
Ile Glu Leu Ala Thr Leu Ala Leu Met Asn Lys Asp Lys Glu Pro Gln
705 710 715 720
Leu Phe Thr Val Tyr Glu Leu Lys Lys Leu Asp Phe Phe Lys His Gly
725 730 735
Tyr Ile Tyr Asn Lys Glu Arg Glu Leu Val Ile Arg Glu Lys Pro Tyr
740 745 750
Lys Ala Ile Gln Asn Leu Ser Tyr Phe Leu Asn Glu Glu Leu Tyr Glu
755 760 765
Lys Thr Phe Arg Asp Gly Lys Phe Asn Glu Thr Tyr Asn Glu Leu Phe
770 775 780
Lys Glu Lys His Val Ser Ala Ile Asp Leu Thr Thr Ala Lys Val Ile
785 790 795 800
Asn Gly Lys Ile Ile Leu Asn Gly Asp Met Ile Thr Phe Leu Asn Leu
805 810 815
Arg Ile Leu His Ala Gln Arg Lys Ile Tyr Glu Glu Leu Ile Glu Asn
820 825 830
Pro His Ala Glu Leu Lys Glu Lys Asp Tyr Lys Leu Tyr Phe Glu Ile
835 840 845
Glu Gly Lys Asp Lys Asp Ile Tyr Ile Ser Arg Leu Asp Phe Glu Tyr
850 855 860
Ile Lys Pro Tyr Gln Glu Ile Ser Asn Tyr Leu Phe Ala Tyr Phe Ala
865 870 875 880
Ser Gln Gln Ile Asn Glu Ala Arg Glu Glu Glu Gln Ile Asn Gln Thr
885 890 895
Lys Arg Ala Leu Ala Gly Asn Met Ile Gly Val Ile Tyr Tyr Leu Tyr
900 905 910
Gln Lys Tyr Arg Gly Ile Ile Ser Ile Glu Asp Leu Lys Gln Thr Lys
915 920 925
Val Glu Ser Asp Arg Asn Lys Phe Glu Gly Asn Ile Glu Arg Pro Leu
930 935 940
Glu Trp Ala Leu Tyr Arg Lys Phe Gln Gln Glu Gly Tyr Val Pro Pro
945 950 955 960
Ile Ser Glu Leu Ile Lys Leu Arg Glu Leu Glu Lys Phe Pro Leu Lys
965 970 975
Asp Val Lys Gln Pro Lys Tyr Glu Asn Ile Gln Gln Phe Gly Ile Ile
980 985 990
Lys Phe Val Ser Pro Glu Glu Thr Ser Thr Thr Cys Pro Lys Cys Leu
995 1000 1005
Arg Arg Phe Lys Asp Tyr Asp Lys Asn Lys Gln Glu Gly Phe Cys
1010 1015 1020
Lys Cys Gln Cys Gly Phe Asp Thr Arg Asn Asp Leu Lys Gly Phe
1025 1030 1035
Glu Gly Leu Asn Asp Pro Asp Lys Val Ala Ala Phe Asn Ile Ala
1040 1045 1050
Lys Arg Gly Phe Glu Asp Leu Gln Lys Tyr Lys
1055 1060
<210> 13
<211> 1106
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.7的氨基酸序列
<400> 13
Met Leu Ile Gln Phe Lys Asn His Tyr Ser Tyr Asn Lys Ser Ile Arg
1 5 10 15
Phe Lys Leu Glu His Lys Asn Gly Lys Leu Pro Lys Leu Glu Ser Asp
20 25 30
Asn Val Asp Leu Asn Lys Leu Val Asp Ile Gly Asn Ser Leu Lys Asp
35 40 45
Ile Phe Glu Glu Leu Val Tyr Thr Lys Asn Asn Tyr Asn Lys Leu Asn
50 55 60
Ser Leu Val Ser Ile Lys Lys Gln Trp Leu Lys Ile Tyr Phe Lys Asn
65 70 75 80
Glu Phe Tyr Ser Asn Gly Lys Ile Gln Asn Tyr Ser Leu Ser Asn Phe
85 90 95
Ser Tyr Leu Pro Asn Lys Leu Ile Glu Trp Leu Asn Asn Trp Gln Asn
100 105 110
Asn Leu Lys Ala Leu Ile Glu Leu Thr Lys Gln Gln Asp Phe Asn Lys
115 120 125
Thr Lys Lys Ser Glu Ile Ala Tyr Ile Leu Ser Leu Phe Asn Gly Lys
130 135 140
Tyr Ser Phe Ser Phe Val Lys Asp Phe Ser Thr Cys Ile Asn His Lys
145 150 155 160
Asn Ser Gln Glu Gln Ile Leu Lys Leu Gln Gly Val Val Glu Asn Phe
165 170 175
Glu Lys Val Leu Asn Leu Cys Ile Gln Glu Tyr Leu Pro Ser Lys Ser
180 185 190
Ala Gly Val Val Ile Ala Gln Gly Ser Met Asn Tyr Tyr Ala Ile Asn
195 200 205
Lys Glu Pro Lys Arg Tyr Asp Asn Ile Leu Ala Asp Leu Asn Gln Lys
210 215 220
Phe Glu Glu Leu Asp Lys Glu Tyr Ile Ala Met Lys Gln Tyr Lys Ser
225 230 235 240
Ser Gln Lys Ser Arg Leu Phe Glu Phe Ile Arg Lys Gly Phe Ser Lys
245 250 255
Asp Gln Ile Leu Ser Glu Phe Lys Lys Lys Glu Asn Asn Glu Val Ser
260 265 270
Phe Val Tyr Asn Asn Gln Ile Ile Ile Arg Ile Tyr Thr Gln Glu Leu
275 280 285
Phe Lys Asp Ser Tyr Cys Leu Gly Glu Val Ile Lys Leu Thr Lys Lys
290 295 300
Ile Glu Glu Leu Asn Glu Ser Lys Asp Ser Asn Asn Asn Leu Pro Glu
305 310 315 320
Glu Thr Lys Lys Glu Ile Thr Lys Leu Lys Lys Glu Ile Gly Phe Tyr
325 330 335
Phe Ile Arg Arg Thr Arg Gly Lys Ser His Asn Asn Tyr Phe Lys Ser
340 345 350
Tyr Tyr Gly Phe Cys Asn Asp Lys Phe Lys Lys Lys Ala Gln Glu Arg
355 360 365
Gly Arg Leu Leu Thr Lys Ile Lys Ala Ile Arg Lys Glu Lys Ile Glu
370 375 380
Ser Gln Asn Leu Arg Tyr Trp Ser Leu Ile Leu Asp Asp Gly Lys Asp
385 390 395 400
Lys Phe Leu Trp Leu Val Pro Lys Glu Asn Met Gln Glu Phe Arg Arg
405 410 415
Glu Leu Ser Lys Ile His Pro Ser Gly Glu Ser Ser Leu Phe Leu Phe
420 425 430
His Ser Leu Thr Met Arg Ala Leu His Lys Leu Cys Phe Ala Gln Glu
435 440 445
Ser Asp Phe Val Lys Glu Met Pro Lys Val Leu Lys Glu Glu Gln Leu
450 455 460
Asn Cys Glu Lys Ala Ser Asn Asp Thr Glu Thr Asn Lys Arg Ile Lys
465 470 475 480
Arg Asn Phe Gly Leu Asn Tyr Ile Lys Thr Lys Asp Glu Leu Thr Leu
485 490 495
Ser Phe Leu Lys Lys Leu Ile Ile Ser Glu Tyr Ala His Glu Arg Leu
500 505 510
Asp Leu Asn His Phe Asp Leu Ser Lys Leu Gln Val Ala Thr Thr Leu
515 520 525
Asn Glu Phe Glu Glu Tyr Leu Glu Asp Ala Cys Tyr Tyr Leu Glu Lys
530 535 540
Ile Ser Ile Ser Ser Ser Met Ile Lys Glu Leu Leu Glu Glu Tyr Asn
545 550 555 560
Ile Leu Asn Phe Arg Ile Thr Ser Tyr Asp Leu Glu Lys Arg Asn Lys
565 570 575
Asn Thr Tyr Gln Thr Pro Glu Ser Asp Ile Lys Arg His Thr Lys Glu
580 585 590
Ile Trp Asn Lys Phe Trp Glu Gly Asp Arg Phe Ile Arg Leu Asn Pro
595 600 605
Glu Ile Lys Ile Arg Tyr Arg Gln Lys Asn Gln Asn Ile Glu Asp Tyr
610 615 620
Leu Lys Glu Lys Gly Phe Asp Leu Thr Lys Ile Lys Asn Arg Phe Leu
625 630 635 640
Gln Glu Gln Tyr Ser Val Ser Phe Thr Phe Ala Leu Asn Ala Gly Lys
645 650 655
Lys Tyr Pro Lys Leu Ala Phe Val Lys Thr Glu Glu Ile Leu Glu Lys
660 665 670
Ile Glu Glu Phe Asn Asp Glu Phe Asn Lys Gln Tyr Phe Asp Asn Ser
675 680 685
Tyr Lys Tyr Gly Ile Asp Arg Gly Asn Ile Glu Leu Ala Thr Leu Cys
690 695 700
Ile Thr Lys Phe Asn Lys Asn Asp Thr Tyr Glu Tyr Lys Gly Lys Lys
705 710 715 720
Tyr Leu Lys Pro Asn Phe Pro Thr Ser Gln Glu Asp Ile Lys Thr Tyr
725 730 735
Glu Leu Lys Asn Glu Trp Tyr Lys Arg Thr Ala Ile Ser Asn Ile Glu
740 745 750
Thr Lys Pro Lys Asn Lys Lys Thr Pro Lys Arg Ile Ile Ala Asn Ile
755 760 765
Ser Tyr Phe Ile Asp Asn Val Glu Asn Glu Glu Trp Phe Asn Lys Lys
770 775 780
Thr Cys Thr Ser Ile Asp Leu Thr Thr Ala Lys Val Ile Lys Gly Lys
785 790 795 800
Leu Ile Leu Asn Gly Asp Val Leu Thr Phe Leu Lys Leu Lys Lys Glu
805 810 815
Ala Ala Lys Arg Ile Leu Phe Glu Leu Val Ala Gln Asn Lys Leu Thr
820 825 830
Ala Lys Asn Lys Glu Leu Lys Trp Lys Ser Asp Asp Gly Asn Asn Ser
835 840 845
Asp Ser Val Arg Leu Ile Cys Asp Val Leu Asp Asn Glu Thr Asn Ser
850 855 860
Ile Tyr Phe Tyr Glu Asp Ser Lys Tyr Gly Arg Gly Phe Glu Gly Leu
865 870 875 880
Leu Thr Thr Asp Lys Thr Ala Tyr Ser Lys Glu Gly Ile Arg Ile Asn
885 890 895
Leu Gln Asn Tyr Leu Asn His Leu Ile Ser Glu Lys Glu Asn Lys Ser
900 905 910
Asn Lys Ala Tyr Ser His Val Pro Ser Ile Glu Lys Ile Asn His Leu
915 920 925
Arg Asp Ala Leu Val Ala Asn Met Val Gly Val Ile Ser Tyr Leu Gln
930 935 940
Ala Tyr Tyr Pro Gly Ile Val Val Leu Glu Asp Leu Asn His Lys Leu
945 950 955 960
Leu Ile Lys His Phe Glu Asp Leu Asn Ile Asn Ile Ser Asn Arg Phe
965 970 975
Glu His Ala Leu Ile Glu Lys Phe Gln Thr Leu Gly Met Val Pro Pro
980 985 990
His Ile Lys Asp Tyr Leu Glu Ile Arg Ser Ser Phe Arg Met Ser Arg
995 1000 1005
Asn Asp Ser Ser Gln Phe Gly Ala Leu Ile Phe Val Ser Lys Glu
1010 1015 1020
Gly Thr Ser Lys Glu Cys Pro Tyr Cys Glu Lys Lys Trp Asn Trp
1025 1030 1035
Gly Lys Glu Lys Glu Ile Glu Leu Lys Phe Ser Lys Lys Gln Tyr
1040 1045 1050
Ile Cys Gly Lys Glu Asn Ser Cys Gly Phe Asp Thr Lys His Ile
1055 1060 1065
Gln Asn Thr Phe Glu Phe Leu Ser Glu Ile Asn Asp Pro Asp Lys
1070 1075 1080
Ile Ala Ala Tyr Asn Ile Ala Lys Arg Gly Phe Lys Ser Phe Ile
1085 1090 1095
Asn Lys Ser Ser Ile Lys Lys Gln
1100 1105
<210> 14
<211> 1085
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.8的氨基酸序列
<400> 14
Met Glu Lys Tyr Lys Ile Thr Lys Thr Ile Arg Phe Lys Leu Leu Pro
1 5 10 15
Asp Lys Ile Gln Asp Ile Ser Arg Gln Val Ala Val Leu Gln Asn Ser
20 25 30
Thr Asn Ala Glu Lys Lys Asn Asn Leu Leu Arg Leu Ile Gln Arg Gly
35 40 45
Gln Glu Leu Pro Lys Leu Leu Asn Glu Tyr Ile Arg Tyr Ser Asp Asn
50 55 60
His Lys Leu Lys Ser Asn Val Thr Val His Phe Arg Trp Leu Arg Leu
65 70 75 80
Phe Thr Lys Asp Leu Phe Tyr Asn Trp Lys Lys Asp Asn Thr Glu Lys
85 90 95
Lys Ile Lys Ile Ser Asp Val Asp Tyr Leu Ser Arg Val Phe Glu Asp
100 105 110
Phe Phe Asn Glu Trp Glu Thr Val Ile Glu Arg Ile Asn Thr Asp Cys
115 120 125
Asn Arg Pro Glu Glu Ser Lys Thr Arg Asp Ala Glu Ile Ala Phe Ser
130 135 140
Ile Lys Lys Ile Ala Thr Lys Gln Met Phe Pro Phe Ile Lys Ser Phe
145 150 155 160
Val Tyr Asn Ser Asn Tyr Lys Asn Ser Glu Glu Thr Lys Ser Lys Leu
165 170 175
Thr Ala Leu Leu Asn Glu Phe Glu Thr Val Leu Lys Ile Cys Glu Gln
180 185 190
Asn Tyr Leu Pro Ser Gln Ser Ala Gly Ile Val Ile Ala Lys Ala Ser
195 200 205
Phe Asn Tyr Tyr Thr Ile Asn Lys Lys Gln Lys Asp Tyr Lys Gly Tyr
210 215 220
Thr Asp Asp Ile Glu Lys Ile Glu Lys Gly Met Asn Ser Lys Phe His
225 230 235 240
Tyr Glu Arg Lys Tyr Asp Gln Leu Leu Glu Glu Leu Asn Leu Ile Ala
245 250 255
Leu Lys Glu Leu Pro Leu Ile Glu Phe Tyr Ser Lys Ile Lys Ser Tyr
260 265 270
Lys Ser Thr Arg Lys Ile Glu Phe Ser Glu Ala Val Ser Lys Gly Leu
275 280 285
Ala Phe Ala Asp Leu Lys Ser Lys Phe Pro Leu Phe Gln Thr Glu Ser
290 295 300
Asn Lys Tyr Ala Glu Phe Leu Glu Leu Thr Gly Arg Ile Thr Gln Ile
305 310 315 320
Ser Thr Ala Lys Ser Leu Leu Ser Lys Asp Asn Pro Glu Ala Gln Lys
325 330 335
Leu Arg Asp Glu Ile Lys Lys Leu Arg Ile Asn Arg Gly Glu Tyr Phe
340 345 350
Lys Asn Asn Phe His Lys Tyr Ile Ser Leu Cys Asn Leu Tyr Lys Lys
355 360 365
Ile Ala Asp Lys Lys Gly Arg Leu Lys Gly Gln Val Lys Gly Ile Glu
370 375 380
Asn Glu Arg Ile Asp Ser Gln Arg Ile Gln His Trp Ala Leu Val Leu
385 390 395 400
Glu Asp Asn Leu Lys His Ser Leu Ile Leu Ile Pro Lys Glu Lys Val
405 410 415
Thr Glu Val Tyr Arg Lys Val Arg Ala Ser Lys Ala Asp Ser Thr Ser
420 425 430
Ser Ser Ser Ser Leu Tyr Tyr Phe Glu Ser Met Thr Tyr Arg Ala Leu
435 440 445
His Lys Leu Cys Phe Gly Val Asn Gly Asn Thr Phe Leu Pro Glu Ile
450 455 460
Gln Lys Glu Leu Pro Glu Tyr Asn Pro Asn Lys Gln Ser Asp Phe Gly
465 470 475 480
Glu Phe Cys Phe His Lys Ser Asn Thr Asp Lys Glu Ile Asp Glu Pro
485 490 495
Lys Leu Ile Ser Phe Tyr Gln Ser Val Leu Lys Thr Asn Tyr Val Lys
500 505 510
Asp Asn Leu Asn Leu Pro Gln Ser Val Phe Asp Glu Ala Thr Val Gln
515 520 525
Thr Phe Glu Thr Arg Gln Asp Phe Gln Ile Ala Leu Glu Lys Cys Cys
530 535 540
Tyr Ala Lys Lys Thr Ile Ile Ser Glu Thr Leu Lys Lys Glu Ile Leu
545 550 555 560
Glu Asp Asn Asn Val Gln Ile Phe Gln Ile Thr Ser Leu Asp Leu Gln
565 570 575
Arg Ser Glu Gln Lys Asn Leu Lys Ala His Thr Lys Ile Trp Asn Arg
580 585 590
Phe Trp Thr Lys Gln Asn Glu Thr Ala Asn Tyr Asp Leu Arg Leu Asn
595 600 605
Pro Glu Thr Ala Ile Val Trp Arg Lys Pro Lys Lys Thr Arg Ile Asp
610 615 620
Lys Tyr Gly Ala Gly Thr Ser Leu Tyr Asp Pro Lys Lys Arg Asn Arg
625 630 635 640
Tyr Leu His Glu Gln Tyr Thr Leu Cys Thr Thr Val Thr Asp Asn Ala
645 650 655
Leu Asn Asn Glu Ile Thr Phe Ala Phe Glu Asp Thr Lys Lys Lys Gly
660 665 670
Thr Glu Ile Val Lys Tyr Asn Glu Lys Ile Asn Gln Thr Leu Lys Lys
675 680 685
Glu Phe Asn Lys Asn Gln Leu Trp Phe Tyr Gly Ile Asp Ala Gly Glu
690 695 700
Ile Glu Leu Ala Thr Leu Ala Leu Met Asn Lys Asp Lys Glu Pro Gln
705 710 715 720
Leu Phe Thr Val Tyr Glu Leu Lys Lys Ser Asp Phe Phe Lys His Gly
725 730 735
Tyr Ile Tyr Asn Lys Glu Arg Glu Leu Val Ile Arg Glu Lys Pro Tyr
740 745 750
Lys Ala Ile Gln Asn Leu Ser Tyr Phe Leu Asn Glu Glu Leu Tyr Glu
755 760 765
Lys Thr Phe Arg Asp Gly Lys Phe Gln Glu Thr Phe Asn Glu Leu Phe
770 775 780
Lys Glu Lys His Val Ser Ala Ile Asp Leu Thr Thr Ala Lys Val Ile
785 790 795 800
Asn Gly Lys Ile Ile Leu Asn Gly Asp Met Ile Thr Phe Leu Asn Leu
805 810 815
Arg Ile Leu His Ala Lys Arg Lys Ile Tyr Glu Glu Leu Ile Ile Asn
820 825 830
Pro Gln Ala Glu Leu Lys Glu Asn Glu Lys Glu Tyr Tyr Leu Tyr Phe
835 840 845
Asp Lys Glu Gly Thr Glu Lys Val Glu Lys Ile Tyr Arg Ser Arg Leu
850 855 860
Asp Phe Glu His Ile Lys Pro Tyr Gln Glu Ile Arg Asn Asp Leu Asn
865 870 875 880
Ala Tyr Phe Lys Asn Val Gln Lys Asn Glu Ala Lys Val Glu Asp Gln
885 890 895
Ile Asn Gln Thr Arg Arg Ala Leu Val Gly Asn Met Ile Gly Val Ile
900 905 910
Tyr Tyr Leu Tyr Gln Lys Tyr Arg Gly Ile Ile Ser Ile Glu Asp Leu
915 920 925
Lys Gln Thr Lys Val Glu Ser Asp Arg Asn Lys Phe Glu Gly Asn Ile
930 935 940
Glu Arg Pro Leu Glu Trp Ala Leu Tyr Arg Lys Phe Gln Gln Glu Gly
945 950 955 960
Tyr Val Pro Pro Ile Ser Glu Leu Ile Lys Leu Arg Glu Leu Glu Lys
965 970 975
Phe Pro Leu Lys Asp Val Lys Gln Pro Lys Tyr Glu Asn Ile Gln Gln
980 985 990
Phe Gly Ile Ile Lys Phe Val Ser Pro Glu Glu Thr Ser Thr Thr Cys
995 1000 1005
Pro Ser Cys Glu Lys Lys Ala Tyr Glu Leu Gln Lys Glu Lys Lys
1010 1015 1020
Gly Glu Glu Lys Pro Ala Glu Asn Lys Arg Tyr Glu Ala Asp Lys
1025 1030 1035
Lys Ala Gly Val Phe Cys Cys Pro Lys Cys Gly Phe His Asn Arg
1040 1045 1050
Thr Asn Pro Met Gly Tyr Glu Ser Leu Asp Ser Asn Asp Lys Val
1055 1060 1065
Ala Ala Phe Asn Ile Ala Lys Arg Gly Phe Glu Asp Leu Gln Lys
1070 1075 1080
His Lys
1085
<210> 15
<211> 1033
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.9的氨基酸序列
<400> 15
Met Glu Asn Ser Asn Leu Tyr Gln Val Val Lys Thr Ile Arg Phe Lys
1 5 10 15
Leu Glu Pro Val Gly Lys Met Asp Thr Pro Lys Phe Gly Asp Lys Asn
20 25 30
Ala Glu Ser Lys Ala Asn Leu Thr Pro Phe Ile Glu Leu Val Lys Lys
35 40 45
Thr Met Thr Asn Val Lys Ala Leu Val Phe Ser Lys Gln Asp Gly Glu
50 55 60
Asp Gly Glu Lys Trp Arg Lys Ile Leu Glu Val Asn Tyr Arg Phe Leu
65 70 75 80
Arg Ser Tyr Leu Lys Asn Ser Phe Tyr Glu Asn Arg Gly Asp Ser Gln
85 90 95
Glu Lys Ser Lys Lys His Lys Ile Ser Asp Leu Glu Tyr Leu Gln Lys
100 105 110
Ala Leu Glu Asn Leu Phe Ala Glu Phe Asp Glu Ile Leu Asp Gly Leu
115 120 125
Glu Asp Phe Glu Lys Arg Asn Thr Lys Asn Gln Tyr Glu Lys Gln Arg
130 135 140
His Ala Gln Ala Gly Leu Leu Leu Asn Arg Leu Cys Lys Arg Ser Asn
145 150 155 160
Phe Gly Phe Leu Lys Ala Phe Val Gly Ala Leu Ala Gln Thr Asn Lys
165 170 175
Pro Phe Phe Asp Asp Lys Thr Asp Lys Leu Lys Lys Gln Ile Asp Lys
180 185 190
Phe Glu Thr Glu Leu Glu Lys Gln Lys Glu Phe Phe Leu Pro Tyr Gln
195 200 205
Ser Asn Gly Val Leu Phe Ala Gly Gly Ser Phe Asn Arg Tyr Ala Ile
210 215 220
Asn Lys Thr Pro Lys Met Leu Asp Lys Glu Leu Arg Glu Glu Gln Thr
225 230 235 240
Asn Leu Lys Lys Ser Leu Cys Glu His Lys Ile Lys Ile Asp Thr Leu
245 250 255
Asn Thr Leu Gly Leu Lys Asn Asp Cys Pro Cys Thr Ser Leu Asp Asn
260 265 270
Ser Tyr Thr Phe Ile Lys Asp Tyr Lys Ala Lys Gln Lys Ser Lys Phe
275 280 285
Ile Glu Leu Val Gln Lys Gly Glu Phe Asp Glu Ala Lys Lys Val Asn
290 295 300
Leu Phe Glu Cys Ser Glu Thr Asp Phe Glu Thr Phe Lys Thr Arg Thr
305 310 315 320
Lys Gln Ile Gln Asn Glu Lys Asp Lys Asp Glu Arg Thr Lys Leu Lys
325 330 335
Gln Lys Arg Gly Glu Phe Phe Lys Ser Gln Lys Arg Gly Lys Phe Phe
340 345 350
Lys Ser Gln Thr Gln Asn Tyr Glu Asn Leu Cys Asp Leu Tyr Lys Lys
355 360 365
Ile Ala Gln Lys Arg Gly Gln Ile Val Ala Lys Ile Cys Ala Ile Lys
370 375 380
Lys Glu Lys Glu Met Cys Glu Gln Val Lys Tyr Trp Cys Val Ala Leu
385 390 395 400
Glu Lys Gly Gly Glu Phe Tyr Leu Tyr Met Phe Leu Arg Asp Glu Asn
405 410 415
Asp Asn Ile Lys Asn Ala Tyr Asp Phe Val Ser Lys Leu Gln Thr Gln
420 425 430
Lys Ser Gly Glu Thr Lys Leu His Tyr Phe Asp Ser Leu Thr Leu Lys
435 440 445
Ala Val Arg Lys Leu Cys Phe Lys Glu Thr Asp Gly Ser Phe Lys Lys
450 455 460
Ala Leu Lys Asn Val Lys Phe Pro Glu Cys Glu Gln Asn Leu Asp Glu
465 470 475 480
Lys Val Lys Ile Ser Phe Tyr Gln Asn Val Leu Lys Asn Ala Lys Thr
485 490 495
Leu Asn Leu Ser Lys Phe Glu Asn Leu Gln Ser Val Thr Glu Gly Lys
500 505 510
Phe Glu Ser Leu Ser Glu Phe Glu Val Ala Leu Asn Met Val Cys Tyr
515 520 525
Thr Lys Thr Val Cys Val Ser Glu Ser Val Glu Lys Glu Leu Lys Lys
530 535 540
Phe Lys Pro Leu Val Phe His Ile Thr Ser Gln Asp Leu Ala Ala Lys
545 550 555 560
Arg Glu Lys Lys Ala His Thr Gln Ile Trp His Glu Phe Trp Arg Glu
565 570 575
Ser Asn Glu Lys Ser Lys Phe Pro Leu Arg Leu Asn Pro Glu Leu Lys
580 585 590
Val Met Trp Arg Glu Ala Arg Pro Ser Arg Val Glu Lys Tyr Ala Glu
595 600 605
Gln Ser Asp Lys Phe Asp Pro Asn Lys Lys Asn Arg Tyr Leu His Pro
610 615 620
Gln Phe Thr Leu Ala Leu Asn Phe Thr Gln Asn Ala His Asn Glu Ala
625 630 635 640
Ile Asn Leu Ala Phe Lys Asp Val Gln Asn Lys Gly Glu Ala Val Lys
645 650 655
Lys Phe Asn Glu Asn Phe Lys Ser Ser Glu Tyr Ala Phe Gly Ile Asp
660 665 670
Val Gly Thr Lys Asp Leu Ala Leu Leu Cys Leu Ile Asp Lys Asn Lys
675 680 685
Lys Pro Val Asn Phe Asp Val Tyr Glu Ile Cys Asn Glu Asn Glu Ile
690 695 700
Cys Asn Glu Lys Leu Gly Phe Glu Lys Phe Gly Phe Tyr Lys Asp Gly
705 710 715 720
Thr Arg Arg Asp Glu Pro Tyr Lys Leu Ile Lys Asn Pro Ser Tyr Phe
725 730 735
Leu Asn Glu Ser Leu Tyr Lys Lys Thr Phe Asn Ala Thr Lys Glu Glu
740 745 750
Phe Glu Arg Ser Phe Ser Glu Leu Phe Lys Arg Lys Ser Val Cys Ala
755 760 765
Leu Asp Leu Thr Thr Ala Lys Val Ile Cys Gly Lys Ile Ile Leu Asn
770 775 780
Gly Asp Phe Ser Thr His Leu Asn Leu Lys Ile Leu Asn Ala Lys Arg
785 790 795 800
Lys Ile Ser Ala Lys Leu Lys Lys Asp Pro Thr Leu Lys Ile Glu Tyr
805 810 815
Asp Asn Asp Asp Asn Ile Leu Phe Gly Ser Asn Val Ile Phe Tyr Tyr
820 825 830
Asn Asn Lys Tyr Glu Ile Val Arg Pro Tyr Asp Glu Ile Lys Asn Glu
835 840 845
Ile Phe Glu Phe His Glu Lys Gln Arg Leu Asp Asp Ala Arg Leu Glu
850 855 860
Asp Asn Ile Asn Lys Thr Arg Ala Asn Leu Val Ala Asn Met Val Gly
865 870 875 880
Val Ile Ser Phe Leu His Lys Glu Phe Ser Gly Phe Val Val Leu Glu
885 890 895
Asn Leu Lys Gln Ser Glu Ile Glu Gly Asn His Arg Leu Lys Phe Glu
900 905 910
Gly Asp Ile Thr Arg Pro Leu Glu Leu Ala Leu Tyr Arg Lys Phe Gln
915 920 925
Ser Lys Cys Leu Thr Pro Pro Ile Ser Glu Leu Ile Lys Leu Arg Glu
930 935 940
Gly Glu Lys Asn Glu Asn Val Glu Ser Asp Leu Ile Leu Gln Phe Gly
945 950 955 960
Ile Ile Lys Phe Val Asp Lys Asp Lys Thr Ser Arg Leu Cys Pro Ala
965 970 975
Cys Gly Lys Asp Ala Tyr Glu Asn Asn Asn Ser Lys Tyr Lys Thr Asp
980 985 990
Lys Lys Asp Gly Val Phe Glu Cys Ala Gly Cys Gly Phe Asn Asn Lys
995 1000 1005
Asn Asn Ala Gly Asp Phe Ala Ala Leu Asp Thr Asn Asp Lys Ile
1010 1015 1020
Ala Thr Phe Asn Ile Ala Lys Arg Gly Leu
1025 1030
<210> 16
<211> 1168
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.10的氨基酸序列
<400> 16
Met Glu Thr Tyr Lys Ile Thr Lys Thr Ile Arg Phe Lys Leu Glu Ala
1 5 10 15
Asp Glu Glu Asn Ser Ile His Ile Lys Glu Asp Ile Ile Asn Ile Glu
20 25 30
Thr Asn Asp Asn Glu Phe Thr Met Val Asp Phe Val Ser Asn Leu Gly
35 40 45
Asn Tyr Ile Lys Asp Leu Lys Asn Tyr Leu Phe Tyr Glu Lys Lys Asp
50 55 60
Gly Ser Leu Ser Phe Lys Asp Lys Ile Ile Ile Lys Asn Glu Trp Leu
65 70 75 80
Arg Gln Tyr Ala Lys Gln Asp Phe Val Glu Leu Lys Ser Lys Lys Arg
85 90 95
Ile Asn Leu Arg Asn Asn Arg Met Glu Gln Ile Lys Ile Gly Asp Ile
100 105 110
Pro Arg Leu Ser Ser Lys Ile Glu Glu Ala Leu Asp Ile Ala Lys Glu
115 120 125
Ile Tyr Ser Lys Leu Ser Asp Asp Ala Thr Leu Glu Gln His Glu Arg
130 135 140
Thr Lys Lys Ala Gln Ile Gly Leu Leu Leu Lys Arg Leu Glu Ala Lys
145 150 155 160
Asn Val Leu Pro Leu Leu Met Asp Leu Val Lys Glu Thr Leu Asp Lys
165 170 175
Asp Glu Thr Asp Asp Leu Ser Ile Arg Leu Lys Arg Gln Ser Gln Lys
180 185 190
Ile Asn Ser Gln Leu Lys Ile Ala Ile Arg Ser Phe Leu Pro Glu Gln
195 200 205
Ser Asn Gly Leu Gln Ile Ala Lys Ala Ser Phe Asn Tyr Tyr Thr Ile
210 215 220
Asn Lys Lys Pro Ile Asp Phe Glu Lys Lys Ile Glu Asp Leu Lys Lys
225 230 235 240
Asn Leu Asn Val Lys Asp Leu Glu Lys Leu Asn Val Tyr Phe Asp Lys
245 250 255
Lys Glu Lys Lys Gln Lys Asn Tyr Leu Gly Lys Lys Ile Phe Ser Leu
260 265 270
Phe Glu Thr Asp Ile Gln Lys Ala Leu Ser Lys Asn Gln Pro Leu Tyr
275 280 285
Leu Gly Asp Ala Pro Met Ile Asp Ser Ala Tyr Val Ser Leu Arg Gln
290 295 300
Ile Phe Lys Lys Ile Lys Ser Glu Gln Lys Lys Gln Phe Ser Glu Leu
305 310 315 320
Met Gln Asn Lys Cys Ser Tyr Asp Glu Leu Lys Asn Ser Asn Leu Tyr
325 330 335
Leu Leu Asn Asp Ile Gly Leu Glu Gln Phe Asn Thr Tyr Arg Glu Lys
340 345 350
Thr Lys Glu Leu Glu Glu Leu Ala Thr Lys Leu Ser Asn Gln Asn Leu
355 360 365
Leu Glu Asn Ala Lys Glu Arg Leu Arg Ser Gln Lys Glu Lys Ile Ala
370 375 380
Lys Glu Arg Gly Asn Ile Met Lys Asp Arg Phe Gln Thr Trp Lys Ser
385 390 395 400
Phe Ala Asn Phe Tyr Arg Thr Val Ser Gln Lys His Gly Lys Ile Leu
405 410 415
Ala Gln Leu Lys Gly Ile Glu Lys Glu Gln Ala Glu Ser Gln Leu Leu
420 425 430
Lys Tyr Trp Ala Leu Ile Cys Glu Lys Glu Asn Gln His Gln Leu Trp
435 440 445
Leu Ile Pro Arg Glu Lys Ala Trp Glu Cys Lys Arg Trp Leu Glu Thr
450 455 460
Val Asn Asp Thr Ser Ile Asp Asn Glu Asn Ser Ile Lys Leu Tyr Trp
465 470 475 480
Phe Glu Ser Leu Thr Tyr Arg Ser Leu Gln Lys Leu Cys Phe Gly Phe
485 490 495
Leu Glu Asn Gly Asn Asn Glu Phe Asn Gln Asn Ile Lys Asp Leu Leu
500 505 510
Pro Lys Asp Arg Ile Gly Asn Thr Ile Asn Gly Glu Phe Ala Phe Glu
515 520 525
Gly Asp Glu Glu Arg Lys Ile Glu Phe Tyr Lys Thr Val Leu Asn Ser
530 535 540
Lys Tyr Ala Lys Gln Val Leu Asn Ile Pro Phe Lys Gln Val Glu Glu
545 550 555 560
Glu Ile Ile Ser Gln Ser Phe Glu Asn Leu Ser Asp Phe Gln Ile Ala
565 570 575
Leu Glu Lys Ile Cys Tyr Arg Arg Phe Ala Ile Tyr Ser Asn Tyr Ile
580 585 590
Ile Ser Phe Asp Ala Gln Ile Phe Asp Ile Thr Ser Leu Asp Leu Lys
595 600 605
Asn Asn Glu Lys Asn Asn Leu Asn Thr His Thr His Ile Trp Arg Asp
610 615 620
Phe Trp Lys Asp Glu Asn Glu Lys Asn Asn Phe Asp Ile Arg Leu Asn
625 630 635 640
Pro Glu Ile Thr Ile Ser Tyr Arg Thr Pro Lys Gln Ser Arg Ile Glu
645 650 655
Lys Tyr Gly Glu Lys Thr Lys Glu Tyr Asp Pro Asn Lys Asn Asn Arg
660 665 670
Tyr Leu His Pro Gln Phe Thr Leu Ile Thr Thr Ile Ser Glu Arg Ser
675 680 685
Asn Ser Gln Thr Lys Thr Leu Ser Phe Ile Glu Asp Glu Asp Phe Lys
690 695 700
Lys Ser Ile Asn Glu Phe Asn Lys Lys Leu Lys Lys Asp Asn Ile Lys
705 710 715 720
Phe Ala Phe Gly Ile Asp Asn Gly Glu Val Glu Leu Ser Thr Leu Gly
725 730 735
Val Tyr Leu Pro Thr Phe Glu Lys Glu Thr His Glu Glu Lys Ile Tyr
740 745 750
Glu Leu Lys Gln Ile Lys Lys Tyr Gly Phe Glu Val Leu Thr Ile Thr
755 760 765
Asp Leu Lys Tyr Lys Glu Thr Asp Tyr Asn Gly Asn Val Arg Lys Ile
770 775 780
Ile Gln Asn Pro Ser Tyr Phe Leu Lys Lys Glu Asn Tyr Ile Arg Thr
785 790 795 800
Phe Ser Lys Ser Glu Gln Glu Tyr Glu Glu Met Phe Ala Lys Leu Phe
805 810 815
Lys Lys Glu His Val Leu Ser Leu Asp Leu Thr Thr Ala Lys Met Ile
820 825 830
Cys Gly His Ile Val Thr Asn Gly Asp Val Pro Ala Leu Phe Asn Leu
835 840 845
Trp Leu Lys His Ala Gln Arg Asn Val Phe Glu Met Asn Asp His Thr
850 855 860
Val Lys Glu Thr Ala Lys Thr Ile Arg Leu Arg Asn Asn Glu Glu Leu
865 870 875 880
Thr Asp Asn Glu Lys Glu Lys Phe Ala Glu Phe Ile Ser Asp Gly Lys
885 890 895
Lys Phe Ala Lys Leu Thr Lys Glu Gly Lys Lys Ser Arg Tyr Leu Lys
900 905 910
Trp Ile Phe Glu Asp Arg Lys Glu Asn Ser Phe Thr Glu Asp Glu Asn
915 920 925
Lys Lys Phe Asn Asp Cys Gln Lys Lys Lys Gly Lys Tyr Asn Ser His
930 935 940
Ile Ile Phe Ala Ser Arg Phe Glu Gly Asp Glu Leu Lys Ser Val Thr
945 950 955 960
Pro Ile Phe Asp Cys Arg His Val Phe Lys Lys Arg Lys Glu Phe Glu
965 970 975
Thr Ile Arg Pro Ile Lys Glu Ile Glu Asn Glu Ile Ser Arg Phe Asn
980 985 990
Thr Asn Arg Thr Ser His Asn Ile Ser Asn Glu Glu Leu Asp Leu Lys
995 1000 1005
Ile Thr Asp Ala Lys Lys Ala Leu Val Ala Asn Ala Ile Gly Val
1010 1015 1020
Ile Asp Phe Leu Tyr Lys Gln Tyr Lys Gln Arg Phe Asn Asp Glu
1025 1030 1035
Gly Leu Ile Ile Lys Glu Gly Phe Asp Thr Gln Lys Val Glu Glu
1040 1045 1050
Asp Ile Glu Lys Phe Ser Gly Asn Ile Tyr Arg Ile Leu Glu Arg
1055 1060 1065
Lys Leu Tyr Gln Lys Phe Gln Asn Tyr Gly Leu Val Pro Pro Ile
1070 1075 1080
Lys Asn Leu Met Ala Val Arg Asn Glu Gly Ile Lys Asp Lys Asn
1085 1090 1095
Ala Ile Leu Arg Leu Gly Asn Ile Ala Phe Ile Asp Pro Ser Gly
1100 1105 1110
Thr Ser Gln Glu Cys Pro Val Cys Lys Glu Lys Ser Lys Glu Lys
1115 1120 1125
His Thr Asn Asn Phe Ile Cys Glu Cys Gly Phe Asn Ser Thr Asn
1130 1135 1140
Ile Met His Ser Asn Asp Gly Ile Ala Gly Phe Asn Ile Ala Lys
1145 1150 1155
Arg Gly Phe Glu Asn Phe Ile Asn Glu Lys
1160 1165
<210> 17
<211> 1187
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.11的氨基酸序列
<400> 17
Met Glu Lys Tyr Lys Ile Thr Lys Thr Ile Arg Phe Arg Leu Asp Ala
1 5 10 15
Asp Asn Thr Ala Ile Ser Ala Ile Val Lys Asp Thr Glu Ala Leu Glu
20 25 30
Ala Arg Gly Gln Gly Phe Lys Ile Lys Lys Phe Val Asn Ala Leu Gly
35 40 45
Arg Phe Leu Ser Gly Asp Gly Val Gln Lys Tyr Leu Tyr Asp Met Ser
50 55 60
Asn Glu Glu Asn Cys Val Phe Lys Arg Asn Leu Val Ile Lys Asn Thr
65 70 75 80
Trp Leu Lys Asn Asn Ala Lys Gln Glu Ile Ala Gly Met Asp Leu Lys
85 90 95
Arg Gly Leu Ile Ile Lys Asp Ile Lys Gly Leu Gln Asp Lys Ile Glu
100 105 110
Glu Ile Tyr Asp Lys Leu Trp Glu Ile Tyr Glu Ile Leu Tyr Glu Ser
115 120 125
Ala Tyr Leu Pro Leu Gln Asp Leu Ala Arg Arg Glu Gly Ile Gly Leu
130 135 140
Leu Leu Lys Lys Leu Ser Val Lys Asn Ala Leu Pro Phe Ile Ile Ser
145 150 155 160
Phe Val Glu Glu Ser Asn Asp Lys Asn Glu Ala Asp Asp Leu Ser Leu
165 170 175
Arg Leu Lys Lys Gln Gly Lys Glu Ile Leu Thr Gln Leu Glu Ile Gly
180 185 190
Ile Asn Glu Tyr Leu Pro Ala Gln Ser Ser Gly Leu Pro Val Ala Lys
195 200 205
Ala Ser Phe Asn Tyr Tyr Thr Ile Asn Lys Thr Pro Val Asp Phe Gly
210 215 220
Glu Lys Ile Gln Glu Leu Glu Lys Arg Leu Ser Val Asp Ile Lys Lys
225 230 235 240
Glu Ile Ser Ser Phe Thr Gly Gly Ile Lys Thr Ala Ile Lys Asn Lys
245 250 255
Ile Ala Gly Lys Lys Ile Leu Leu Gly Asp Thr Pro Met Phe Glu Ser
260 265 270
Glu Asn Ser Val Ser Leu Arg Gln Ile Leu Lys Asn Ile Lys Ser Glu
275 280 285
Gln Lys Ala Gln Phe Asn Lys Phe Met Thr Thr Gln Asn Asn Pro Gln
290 295 300
Leu Glu Glu Met Lys Thr Met Gly Trp Tyr Leu Phe Gly Asp Ile Thr
305 310 315 320
Glu Gly Glu Phe Asn Asp Tyr Lys Glu Gln Thr Lys Glu Ile Glu Arg
325 330 335
Val Gly Ala Lys Ile Asn Gln Cys Gly Asn Ile Lys Glu Lys Lys Glu
340 345 350
Leu Arg Ser Gln Leu Gln Lys Leu Lys Lys Lys Arg Gly Glu Leu Ile
355 360 365
Ser Glu Ala His Lys Lys Gly Gly Asn Asp Lys Asn Phe Lys Thr Tyr
370 375 380
Lys Glu Phe Ala Lys Phe Tyr Arg Lys Ile Ala Gln Arg His Gly Lys
385 390 395 400
Ile Leu Ala Gln Ile Lys Gly Ile Glu Lys Glu Lys Ile Asp Ser Ala
405 410 415
Met Leu Asn Tyr Trp Ala Ala Val Ile Glu Leu Ser Gly Arg His Lys
420 425 430
Leu Val Leu Ile Pro Lys Lys Asp Glu Asn Ala Lys Lys Cys Ile Glu
435 440 445
Trp Leu Glu Asp Glu Ser Lys His Lys Asn Gly Ser Cys Lys Ile Phe
450 455 460
Trp Phe Glu Ser Phe Thr Phe Arg Ser Leu Gln Lys Leu Cys Phe Gly
465 470 475 480
Asn Leu Asp Ser Gly Thr Asn Thr Phe Asn Gln Lys Ile Gln Asn Leu
485 490 495
Leu Pro Cys Asp Glu Arg Gly Asn Leu Met Asn Gly Glu Phe Ala Phe
500 505 510
Lys Gly Asp Glu Gln Glu Lys Ile Lys Phe Tyr Lys Lys Val Leu Gln
515 520 525
Ser Gln Lys Asp Ile Asn Leu Pro Gln Lys Glu Val Val Asp Asn Val
530 535 540
Val Gly Arg Lys Phe Glu Thr Met Asp Glu Phe Lys Ile Ala Leu Glu
545 550 555 560
Glu Ile Cys Tyr Ile Arg Arg Glu Arg Leu Ser Ala Asn Ala Glu Ser
565 570 575
Glu Leu Lys Ser Lys Phe Asn Ala Gln Ile Phe Asp Ile Thr Ser Leu
580 585 590
Asp Leu Arg Asn Pro Val Asn Cys Ala Gly Lys Pro Glu Val Tyr His
595 600 605
His Asn Asp Lys Arg His Thr Glu Ile Trp Lys Glu Phe Trp Ser Leu
610 615 620
Asp Asn Glu Arg Arg Asn Phe Asn Ile Arg Leu Asn Pro Glu Ile Thr
625 630 635 640
Ile Thr Tyr Arg Lys Pro Lys Glu Ser Arg Ile Leu Lys Tyr Gly Lys
645 650 655
Gly Thr Glu Lys Tyr Asn Ala Asp Met Lys Asn Arg Tyr Leu Tyr Pro
660 665 670
Gln Tyr Thr Leu Leu Thr Thr Ile Ser Glu His Cys Asn Thr Pro Thr
675 680 685
Lys Ile Leu Ser Phe Met Thr Asp Asn Glu Tyr Glu Glu Ser Ile Lys
690 695 700
Ala Phe Asn Ser Lys Leu Lys Lys Glu Asp Ile Lys Phe Ala Phe Gly
705 710 715 720
Ile Asp Ser Gly Glu Thr Glu Leu Ser Thr Leu Gly Val Tyr Leu Pro
725 730 735
Glu Phe Ser Ala Glu Ser Thr Glu Leu Lys Asp Ile Glu Lys Tyr Gly
740 745 750
Phe Asn Val Leu Thr Ile Lys Asp Leu Asn Tyr Thr Glu Thr Asp Tyr
755 760 765
Asn Gly Ser Asp Lys Lys Ile Val Lys Asn Pro Ser Tyr Phe Val Asp
770 775 780
Lys Ser Leu Tyr Met Arg Thr Phe Lys Lys Thr Glu Gln Glu Tyr Glu
785 790 795 800
Lys Met Phe Ala Glu Gln Phe Glu Ala Lys Lys Arg Leu Ser Leu Asp
805 810 815
Leu Ser Ala Ala Lys Val Ile Cys Gly His Ile Val Thr Asn Gly Gly
820 825 830
Val Ser Glu His Phe Gly Leu Trp Leu Lys His Ala Gln Arg Thr Ile
835 840 845
Phe Trp Met Asn Asp His Thr Glu Lys Lys Thr Ala Lys Asn Ile Lys
850 855 860
Leu Lys Asp Ser Ser Glu Leu Thr Tyr Asp Glu Arg Glu Lys Phe Ala
865 870 875 880
Glu His Ile Ser Ser Asp Glu Lys Phe Lys Lys Leu Asp Val Glu Glu
885 890 895
Lys Lys Arg Tyr Val Arg Trp Ile Phe Glu Asp Arg Glu Thr Leu Asn
900 905 910
Phe Thr Glu Ala Glu Asn Lys Lys Phe Gly Gly Tyr Gln Lys Lys Lys
915 920 925
Gly Asp Tyr Arg Leu Gly Ile Leu Phe Ala Ser Cys Phe Ile Gly Lys
930 935 940
Glu Leu Glu Ser Val Thr Gln Ile Leu Asp Cys Arg His Ile Phe Lys
945 950 955 960
Lys Arg Glu Glu Phe Tyr Ser Leu Lys Ser Lys Glu Asp Ile Glu Ala
965 970 975
Glu Ile Lys Arg Tyr Asn Thr Asp Tyr Thr Asn His Asn Ile Ser Thr
980 985 990
Glu Gln Leu Asp Leu Lys Phe Val Asn Val Lys Asn Ala Leu Val Ala
995 1000 1005
Asn Ala Val Gly Val Ile Asp Leu Leu Tyr Lys Gln Tyr Lys Glu
1010 1015 1020
Arg Leu Gly Gly Glu Gly Leu Ile Ala Lys Glu Gly Phe Asp Thr
1025 1030 1035
Lys Lys Val Glu Glu Asp Met Glu Lys Phe Ser Gly Asn Ile Tyr
1040 1045 1050
Arg Ile Leu Glu Arg Lys Leu Tyr Gln Lys Phe Gln Asn Tyr Gly
1055 1060 1065
Leu Val Pro Pro Ile Lys Asn Leu Met Ala Val Arg Ala Asp Lys
1070 1075 1080
Val Glu Ile Ser Glu Ala Glu Lys Ser Lys Ile Arg Glu Asn Cys
1085 1090 1095
Lys Ile Ser Lys Ile Asp Pro Glu Asn Glu Ile Ile Lys Arg Asn
1100 1105 1110
Lys Thr Leu Ile Leu Arg Leu Gly Ser Ile Ala Phe Val Asn Asp
1115 1120 1125
Ala Asp Thr Ser Gln Glu Cys Pro Ala Cys Gly Thr Lys Ser Lys
1130 1135 1140
Glu Lys His Val Asp Asn Phe Ile Cys Gly Cys Gly Phe Asn Ser
1145 1150 1155
Thr Gly Ile Ile His Ser Asn Asp Gly Val Ala Gly Phe Asn Ile
1160 1165 1170
Ala Lys Arg Gly Phe Val Asn Leu Met Glu His Glu Leu Arg
1175 1180 1185
<210> 18
<211> 1195
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.12的氨基酸序列
<400> 18
Met Glu Lys Tyr Lys Leu Thr Lys Thr Ile Arg Phe Lys Leu Lys Pro
1 5 10 15
Lys Asp Ile Ser Ala Ile Lys Arg Asp Val Glu Ala Leu Glu Gln Gln
20 25 30
Lys Phe Asp Leu Val Leu Phe Val Tyr Asn Leu His Asn Phe Ile Gly
35 40 45
Lys Leu Lys Glu Tyr Leu Phe Phe Gln Lys Glu Lys Asp Glu Phe Val
50 55 60
Ile Lys Asp Lys Leu Thr Ile Lys Lys Thr Trp Leu Lys Gln Tyr Ala
65 70 75 80
Lys Gln Glu Ile Ala Gly Leu Glu Leu Asn Arg Glu Gln Thr Leu Gly
85 90 95
Asn Ile Lys Gly Val Ser Ala Arg Ile Glu Arg Ala Val Asp Asp Val
100 105 110
Asn Lys Ile Tyr Val Glu Leu Ala Met Glu Ala Lys Leu Asn Glu Arg
115 120 125
Ala Lys Lys Ala Lys Thr Glu Gln Leu Ile Lys Arg Leu Asp Thr Arg
130 135 140
Asn Ala Leu Pro Leu Leu Val Ser Leu Ile Glu Gln Ser Ser Asp Lys
145 150 155 160
Tyr Glu Thr Gly Asn Leu Ser Ile Gln Leu Lys Arg Leu Gly Lys Arg
165 170 175
Leu Gln Thr Gln Leu Leu Ser Gly Ile Lys Lys Tyr Leu Ala Glu Gln
180 185 190
Ser Asn Gly Leu Pro Ile Ala Lys Ala Ser Phe Asn Tyr Tyr Ala Ile
195 200 205
Asn Lys Lys Pro Val Asp Tyr Ile Asp Lys Ile Lys Gln Leu Gln Lys
210 215 220
Asp Leu Glu Ile Lys Lys Asn Arg Arg Ser Glu Glu Arg Tyr Asp Lys
225 230 235 240
Lys Lys Arg Lys Asn Ile Lys Ile Phe Asn Asp Ser Lys Leu Trp Ile
245 250 255
Lys Ile Lys Lys Asp Ile Glu Lys Glu Arg Gly Asn Lys Thr Leu Ile
260 265 270
Leu Gly Tyr Ala Pro Met Ile Glu Pro Gly Asn Tyr Val Tyr Leu Arg
275 280 285
Gln Ile Leu Lys Asn Ile Lys Leu Glu Gln Lys Asn Lys Phe Ser Lys
290 295 300
Leu Met Gln Ser Lys Ser Leu Thr Phe His Asp Leu Asn Asn Asn Asn
305 310 315 320
Gln Leu Tyr Leu Phe Lys Asp Ile Leu Glu Gly Glu Phe Asn Lys Tyr
325 330 335
Lys Gln Lys Thr Asn Glu Ile Glu Thr Lys Ala Glu Lys Arg Asn Gln
340 345 350
Cys Asn Asn Asp Glu Leu Lys Arg Lys Leu Asn Ser Glu Leu Gln Gln
355 360 365
Leu Arg Lys Asp Arg Gly Ser Leu Ile Asn Ala Ala Asp Gly Arg Pro
370 375 380
Lys Gly Arg Phe Lys Thr Tyr Lys Tyr Phe Ala Asn Phe Tyr Arg Asn
385 390 395 400
Val Ala Gln Lys His Gly Arg Ile Leu Ser Thr Leu Lys Gly Ile Glu
405 410 415
Lys Glu Met Val Glu Ser Gln Leu Leu Lys Tyr Trp Thr Ile Ile Thr
420 425 430
Glu Glu Asn Asn Gln His Ser Leu Val Leu Ile Pro Lys Glu Arg Ala
435 440 445
Gly Glu Tyr Lys Lys Asp Leu Glu Asn Ser Ile Pro Ser Asp Pro Ser
450 455 460
Ser Lys Ile Lys Val Tyr Trp Phe Glu Ser Phe Thr Leu Arg Ser Leu
465 470 475 480
Arg Lys Leu Cys Phe Gly Tyr Val Asn Asn Asn Thr Gly Ser Asn Thr
485 490 495
Phe Tyr Pro Glu Leu Lys Lys Ser Asp Glu Leu Arg Lys Tyr His Asp
500 505 510
Glu Arg Gly Asn Phe Ile Lys Gly Glu Phe Tyr Phe Lys Gly Asp Glu
515 520 525
Gln Lys Ile Ile Gln Phe Tyr Lys Asp Val Leu Arg Ser Asn Tyr Ala
530 535 540
Gln Lys Val Leu Lys Phe Pro Lys Gln Gln Val Lys Asp Glu Leu Ile
545 550 555 560
Gly Arg Glu Phe Ser Ser Leu Asp Glu Phe Gln Ile Ala Leu Glu Lys
565 570 575
Ile Cys Tyr Gln Arg His Val Val Cys Ser Gln Lys Val Val Asp Ala
580 585 590
Leu Ser Arg Tyr Asn Ala Gln Ile Phe Leu Ile Thr Ser Leu Asp Leu
595 600 605
Gly Asn Pro Ala Asn Cys Val Asp Lys Pro Lys Gln Phe Ser His Phe
610 615 620
Asp Lys Lys His Thr Arg Ile Trp Lys Glu Phe Trp Ser Ser Lys Asn
625 630 635 640
Glu Thr Ala Asn Phe Asp Ile Arg Leu Asn Pro Glu Ile Val Ile Thr
645 650 655
Tyr Arg Gln Pro Lys Gln Ser Arg Ile Lys Lys Tyr Gly Pro Glu Ser
660 665 670
Thr Arg Tyr Asp Asp Arg Lys His Asn Arg Tyr Leu Tyr Pro Gln Phe
675 680 685
Thr Leu Ile Thr Thr Ile Ser Glu Tyr Ser Asn Ala Pro Thr Lys Ala
690 695 700
Leu Ser Phe Leu Thr Asp Glu Glu Phe Lys Gly Ala Val Asp Glu Phe
705 710 715 720
Asn Lys Lys Phe Lys Lys Glu Asn Ile Arg Phe Ser Leu Gly Ile Asp
725 730 735
Asn Gly Glu Thr Glu Leu Ser Thr Leu Gly Val Tyr Leu Pro Val Phe
740 745 750
Lys Lys Asp Ser Asn Glu Lys Val Val Ala Glu Leu Lys Lys Val Asn
755 760 765
Lys Tyr Gly Phe Asn Phe Leu Thr Ile Lys Asp Leu Ser His Val Glu
770 775 780
Lys Asp Lys Asn Gly Arg Val Arg Lys Ile Ile Gln Asn Pro Ser Tyr
785 790 795 800
Phe Leu Ser Lys Glu Gln Tyr Met Arg Thr Phe Gly Arg Thr Glu Gln
805 810 815
Glu Tyr Asn Asn Met Phe Ala Glu Gln Phe Glu Glu Lys Ala Phe Leu
820 825 830
Ser Leu Asp Leu Thr Thr Ala Lys Val Ile Asn Gly His Ile Val Thr
835 840 845
Asn Gly Asp Val Pro Thr Phe Leu Asn Leu Trp Met Arg His Ala Gln
850 855 860
Arg Asp Ile Trp Asp Met Asn Asp His Thr Lys Glu Lys Thr Ala Lys
865 870 875 880
Lys Ile Val Ile Lys Asn Asn Asp Glu Leu Thr Asp Ala Glu Lys Val
885 890 895
Lys Phe Val Glu Tyr Ile Ser Asp Glu Thr Asn Tyr Ala Lys Leu Asn
900 905 910
Phe Asn Glu Lys Lys Arg Tyr Val Leu Trp Ile Phe Glu Asn Arg Lys
915 920 925
Asn Ile Asn Phe Thr Asp Ala Glu Lys Lys Lys Phe Glu Pro Cys Gln
930 935 940
Lys Arg Lys Gly Asn Phe Ser Lys Asp Ile Leu Phe Ala Val Cys Tyr
945 950 955 960
Ile Gly Ser Glu Ile His Ser Val Thr Asn Ile Phe Asp Val Arg Asn
965 970 975
Ile Phe Lys Met Arg Lys Asp Phe Tyr Val Leu Lys Ser Glu Met Glu
980 985 990
Ile Lys Lys Glu Ile Glu Ser Tyr Asn Thr Thr Ala Gly Ile Gln Glu
995 1000 1005
Ile Ser Asn Glu Glu Leu Asp Leu Lys Ile Asn Arg Leu Lys Gln
1010 1015 1020
Ala Val Val Ala Asn Ala Val Gly Val Ile Asp Tyr Leu Tyr Ile
1025 1030 1035
Tyr Tyr Lys Lys Lys Thr Gly Gly Glu Gly Leu Ile Ile Lys Glu
1040 1045 1050
Gly Phe Asp Thr Lys Lys Val Ala Lys Ala Leu Glu Lys Phe Ser
1055 1060 1065
Gly Asn Ile Tyr Arg Ile Leu Glu Arg Lys Leu Tyr Gln Lys Phe
1070 1075 1080
Gln Asn Tyr Gly Leu Val Pro Pro Ile Lys Ser Leu Met Ala Val
1085 1090 1095
Arg Glu Glu Gly Ile Glu Asn Asn Lys Asp Ala Ile Leu Arg Leu
1100 1105 1110
Gly Asn Val Gly Phe Ile Asp Pro Thr Gly Thr Ser Gln Gln Cys
1115 1120 1125
Pro Val Cys Ser Lys Gly Lys Leu Asn His Thr Thr Lys Cys Ser
1130 1135 1140
Lys Asn Cys Gly Phe Asn Ser Lys Asn Ile Met His Ser Asn Asp
1145 1150 1155
Gly Ile Ala Gly Tyr Asn Ile Ala Lys Arg Gly Phe Glu Asn Phe
1160 1165 1170
Ile Ser Gln Lys Lys Gly Tyr Asp Val Ile Asn Asn Gly Thr Lys
1175 1180 1185
Tyr Asn Asn Leu Lys Ser Gln
1190 1195
<210> 19
<211> 2649
<212> DNA
<213> 人工序列
<220>
<223> Cas12g.1编码核酸序列
<400> 19
atgctgtaca ccatgaacgt gaagaccatc aagctgaagg tggacgccac caaggaggtg 60
gagtccaggc tgaccaagat gctgctggtg cacaacaaca tcggcaggga gatcatcaac 120
ttcctgatcc tgtgctccgg caacgacaac atcaggaaga ccaagttcga cgagttcggc 180
aactcctacg acgagttctg caacctgaag ctggaccagt tcaacctgta cgacaggctg 240
accgagatcc acgacgaggt gaccctggag gacttccaga agaccctgaa cgacatctac 300
gacctggtgc tgaactccaa gtccttctcc aacgtgtcct ccaccatctt caacaagaac 360
aagaaggtga acttcgacga gaccaagaag ggcgacctgt ccaggaagtg cctgatgaac 420
gccagggact ggggcgtgct gccgctgatc tccgtggacg acgacatcgt gacctgcggc 480
accctgaagg gcatcctgtc cgagtgccag tccaggatcc tgtcctggaa cgagtgcaac 540
ctgtccacca aggagaccta ctccgagaag aagtccgagt accagtccat cctggacgac 600
tccatgacca aggacgccga cgtgaccacc gccatgatcc agttcatgga cgacgtgtcc 660
aacgtgtacg gctccaacaa cgagaaccag ctgaagtggt tcaacaacag gttcctgacc 720
tacgtgagga acaagatcag gccgttcctg ctgaccaact ccccgatcga caacttcgag 780
cagtccgaca cctcctacaa ctgctccatc gagatcgtga ggatcctgtc caagtacgag 840
atcctgtgga aggacgaggt gtccgtgaac aggtacaaga agacctgcga cgacggcatc 900
aacatcgaga agtacaggta cctggtgcac gccaagtccg acttcctgag gtacaaggag 960
accgcctcct tcaaggagat ccacgccgtg aagtccccga tctccctgtg cttcggcaac 1020
aactaccagc cgttctccct gtccgacgtg ggcgacaggc acaacatcaa cttcggctac 1080
aagttcggca agctgggcaa gcagaggaag gagtgctcct tcaacctgaa ctacaggagg 1140
aagaaggtga agtacgccaa caccccggtg aggtccgacg agaacaagtg ctacctggac 1200
aacctggaga tcgaggacgc caagaacggc tcctacaagc tgtcctacat ggtgaacaag 1260
aagtacaaga gggagtcctt catcaaggag ccgaagatga agatgtacaa cggcaagctg 1320
tacatgtact tcccgatgtc caacgagttc gaggaggaca gggactcctt cgccctgctg 1380
acctacttct ccaggtcctc caactccaag tcccagatcg acgaggcctc caacatcctg 1440
cagaacagga agatcagggt gtgcggcgtg gacctgggca tcaacccgac cttcgccctg 1500
tccgtgctgg agtactccga caacaagatc accgacacca acatcggcat gaagcacgag 1560
ggctcctaca acaacttctc cgagatcagg aagcagatca acgacgtgac cgacatgatc 1620
tcctacctga agtccaagta cgacaactgc gagaaggact actcctccaa gatcgacgac 1680
cacatcaagt ccaggctgaa cgaggagatc tccaacttct gcgacctggt gtcctacaag 1740
aggaacaaga acaccatcat caggaaggag atcaagaacg tggagaagga gatcaacaag 1800
atcaagaact gcaggaggca caccctgaag aaggacctga ccgagaactt cggctgggtg 1860
tccgccctga acgagttcat ctccctgaag cactccttca acgacatggg cgagtccttc 1920
gactccaaga ccaacccgtc ctactcctac ttcgagaagt ggaagaggta catcgacaac 1980
atcaaggacg actccctgaa gaccgtgtcc agggagatcc tgaacttctg catcgagaac 2040
tccgtggact tcatcgccct ggaggacctg cagaccttcg ccccgtccga cgacaggacc 2100
aagtcccaca acaagctgac ccagctgtgg tgcttcggca agctgaagaa gtgcctggag 2160
gacatcgcct ccatgtacgg catccacgtg tactcctcca ccgacccgag gaacacctcc 2220
gacacccact tcgagtccaa gaacttcggc tacagggacg agtccaacaa gcacaacctg 2280
tgggtgaacg tggacggcga gtacaccgtg gtggactccg acatcaacgc ctccaagaac 2340
atcgccaaca ggttcctgac ccaccacaag gacctgaagc agctgccgat gatcggcgac 2400
ggcaccctgt tcaagatcga ctcctcctcc aagaggaaca agtccttcgc cgtgaagctg 2460
aacatccaca agaacgtgta cgagctgatc gacggcgagt tcgtgaagtc caacaagaag 2520
ccgaacggca cctccaggaa gcagaccgcc tacatccacg gcgacatgtt catcgactcc 2580
atctcccaca agaacaagaa gatgttcctg agggagaacc tgatcaggaa cggcttcatc 2640
tccaagtga 2649
<210> 20
<211> 2808
<212> DNA
<213> 人工序列
<220>
<223> Cas12g.2编码核酸序列
<400> 20
atgaacaaga ccgacaccca gaacaacgag cagatcaaca agccgaccca gctgctgaac 60
aacaaggaca tcgagctgac cgtgaagacc gtgaagtccg ccaccgtgaa ggtggacaac 120
aactccaaga aggagctgtt cggcctgttc aactacttca cctccgtggc ctccggcatc 180
aaggacaagg tgtacaacct gcagtccgac gagaagaccg ccccgatctt caacgactac 240
gtgaagcagc cgcagagggg caggtccgcc gccaccaccc tgttcaccaa gctggacgcc 300
gagaagacct acacctccca gcactccttc ccgggcaagt ggagggactc cggcatcttc 360
ccgctgtaca acaaggagtc cgagaagtac gacctgtcca cccacggcta ccactactcc 420
gccaacgccg agatccacac ccagctggac tcccacgacg agtgcaacaa ggagtgcgag 480
aaggagtacg ccgccctgag ggacgaggtg aacaactaca agtacgagtt caccctgcag 540
ttcaaggccg agaacgccga gaagttctac aacttcgtgg agaagctgac cctgatgggc 600
tggaggtacg acgccacctt caggtccttc ttcgagctgc acatgcaccc gaagctgaag 660
accggcgaga ccacctacag ggccacctac aagctgccgt ccggcaagtc caagaggtac 720
tccttcttca gggacgacat cgccgacgag atcgccaaga acccggagtt ctggccgatg 780
ctggagtcct ccaacgccat ctcctggatc aactccaaca acctgctgtc caggaagaag 840
gacaaggcca actactcctc cacctccctg atcaagtccc agatcaggct gtacctgggc 900
aacaacggcg tgccgttcac cgccagggag cacgacggca ggatctactt ctccttcagg 960
ctgccggcca tcaacggcga gaagggcagg atggtggaga tcccgtgctc ctacaagaag 1020
gtgttcaacg gcaaggccag gaagtcctgc tacctgggcg gcctgaccat cgagaagacc 1080
gacgccggca agcacatctt caagtactcc gtgaacaaca agaagccgca ggtggccgag 1140
ctgaacgagt gcttcctgag gctggtggtg aggaacaggg agtacttcaa caacgtggtg 1200
gccggcaaga tcaccgacat caacaccgac cacttcgact tctacgtgga cctgccgctg 1260
aacgtgaagg aggacccgat ccacgacctg tcctccaccg aggtgttcgg caagaacggc 1320
ctgaggtcct actactcctc cgcctacccg gagatcaaga acctgggctc ccagatcgag 1380
accggcaaga acctgacctg cccgatcacc aagacccaca acatcatggg catcgacctg 1440
ggccagagga acccgttcgc ctactgcatc aaggacaaca ccggcaagct gatcgcccag 1500
ggccacatgg acggctccaa gaacgagacc tacaagaagt acatcaactt cggcaaggag 1560
tccacctccg tgtcccacct gatcaaggag accaggtcct acctgcacgg cgacccggag 1620
gccatctcca aggagctgta caacgaggtg gccggcttct gcaacaaccc ggtgtcctac 1680
gaggagtacc tgaagtacct ggactccaag aagttcctga tcaacaagga ggacctgtcc 1740
aagaacgcca tgcacctgct gaggcagaag gaccacaact ggatcggcag ggactggctg 1800
tggtacatct ccaagcagta caagaagcac aacgagaaca ggatgcagga cgccgactgg 1860
aggcagaccc tgtactggat cgactccctg tacaggtaca tcgacgtgat gaagtccttc 1920
cacaacttcg gctccttcta cgacaagaac ctgaagaaga aggtgaacgg caccgtggtg 1980
ggcttctgca agaccgtgca cgaccagatc aacaacaaca acgacgacat gttcaagaag 2040
ttcaccaacg agctgatgtc cgtgatcagg gagcacaagg tgtccgtggt ggccctggag 2100
aagatggact ccatgctggg cgacaagtcc aggcacacct tcgagaacag gaactacaac 2160
ctgtggccgg tgggccagct gaagaccttc atggagggca agctggagtc cttcaacgtg 2220
gccctgatcg agatcgacga gaggaacacc tcccaggtgt gcaaggagaa ctggtcctac 2280
agggaggccg acgacctgta ctacgtgacc gacggcgagt cccacaaggt gcacgccgac 2340
gagaacgccg ccaacaacat cgtggacagg tgcatctcca ggcacaccaa catgttctcc 2400
ctgcacatgg tgaacccgaa ggacgactac tacgtgccga cctgcatctg ggacaccacc 2460
gaggagtccg gcaagagggt gaggggcttc ctgaccaagc tgtacaagaa ctccgacgtg 2520
gtgttcacca agaagggcga caagctggtg aagtccaaga cctccgtgaa ggagctgaag 2580
aagctggtgg gcaagaccaa ggagaagagg ggccagtact ggtacaggtt cgagggcaag 2640
tcctggatca acgaggccga cagggacacc atcatcctga acgccaagaa gatctccagg 2700
gagagggaca acggcgagca gtccaccgac accaggtccc agaacgtgac cgtgtccgtg 2760
ctggacgtgt gcgagaccgc cgagaagaag aagctggtgc tggtgtga 2808
<210> 21
<211> 2664
<212> DNA
<213> 人工序列
<220>
<223> Cas12h.1编码核酸序列
<400> 21
atggccctga tccagagggc cggcgtgctg aagaccaagt ccgacttccc gaaggtgatc 60
aaggactggc acgactccct gctggccgac tacaggaagt tcttcccgat catcttctcc 120
tggtgcccgg agtacggcta caccaccatc caggacaaca agccggtgtt cgtgtccccg 180
gaggagagga tggagtccat caggaaggag gccaaggagc acctgaacga ggtgctggcc 240
ttcggcaaga tgatcggctc caagggcgtg ggcggctcct cctcctacgc catcttctac 300
aagcaccaca agaacaacga gaacggcgcc tacaccccgt ccagggccaa gttcatgaag 360
gagggcatcc acaacaggag ggtggagctg gtggacgtgc tgatgctgaa cgccatcccg 420
gacgaggagt gggtgaagat cgcccaggag gtggtgggct actccgagga gaggctgaag 480
ctgtactgga acaagttcat cgccaagagg gtggtgtccc acgacaggaa gctgggcaag 540
atcgtgaggg agaagtacct ggagccgaag ggcctggtgt gcgcccagcc ggagaactcc 600
acctactgca gggtgctgac cgagatcatc aagaggcagc tgcactccca gatcgagaag 660
tccaagttcc acgaggagga gctgaagtcc atcgagaaga ccgtgtccga gttcgactcc 720
ccgctgctgg acttcatctg ccagtacgcc gaggagctga accagatcaa ctccggcctg 780
tccaagtacg tgatcaagaa cgccgtgaag gaggtgatct ccccgccgga gaagcagtcc 840
gagatctacg tgcagtccca ggtgctgtcc caggagaagt acaagccgct ggtgaacgcc 900
accatcaagg agatcctgtc cggctacgag cagtggaagg tgaagtccag gtacgagaac 960
aggctgaaga acaggaagta cgtgctgtac ccgaagctgt ccgccaacta caagatcccg 1020
atcggccaga actccctggg caagttcaag atcaacgtgt ccgagaacgg cgagatcgtg 1080
atcaggctga acgacatggc cgacgtggtg tgcatgccgt ccaagtactt cttcaacctg 1140
aagtcctccc cggtggtgga caagaagaag cagctggtgg gctaccagat ctccttcaac 1200
cacaactcca ggaggaagga gccgaccgag aagccggact tcaacggcat cgtgaaggag 1260
atcggcctgc agctgaagga cgacggcagg ttctacatca ccctgccgta ctgcatggag 1320
tactccaacg acaacttcga cctgatcagg ccgctgctga cctcctcccc gaccgaggac 1380
cagatcaaga agatgccgtc cgagttcaac gtggtgggct tcgacctgaa cctgtccatg 1440
ccgctgccga tcaccagggc catcgtgggc aagtccgtga agggcgagat caacgtggag 1500
tacctgggcc aggccaaggt gatcgagtcc acccacctga tctacgacaa caacaggtgc 1560
aaggtgctga tcgcctacaa gaggcagtgc gacctgatca agagggccat cagggagtgg 1620
aagatctgca agggcaagaa catcgacatc tccgagaaga cctacgagtg gctggagtcc 1680
cacaccaaga ggtggaaccc gtccaggcag ccggagtcca tgcaggacag gttctccgtg 1740
tccaagatga ggatccagat cctggtgaac aaggccaagt ccaggatcgc caagtacaac 1800
gacaactcct ggaagaccgg ccacggcaac gagtccgagc tgatcaggct gatcgacgcc 1860
gacgacgcct acaactccct ggtgtccacc tacaacagga tccacctgaa gtccaaccag 1920
ttcatctacg ccctgccgtc caagaacaac tccaggtcca acaagaagga gtactgcctg 1980
aggaggatcg ccgccaagat cgccaggtac tgccacctgc acaacgtgaa catctgcatc 2040
ggcgagaacc tgtccttcca gcaggactcc gacaacatct ccaaggacaa ctccctggtg 2100
aggctgttct cctccaagtc catcgccaac tacatgaagc tggccatgga gaagttcggc 2160
atcgccttca tcgactccgc cgacccgtcc ggcacctcca agaccgaccc ggtgaccggc 2220
aacatcggct acaggaacaa gttcgacaag aggaagctgc acgtgatcag gaacggcaac 2280
tggggctggg tggactccga catcgccgcc tccctgaaca tcctgatcag gggcatcaac 2340
aggtccatcg tgccgtacaa gttcttcgtg ggcaagaaga agcaggagtc caagaggctg 2400
aaccacttcc tgaacaagat cttcggcacc accaaggtgt tcttctacga ggaccagttc 2460
ggcttcgcca acccgtccct gtccaagaag gagggcgaga acctgatcgc caaccagtac 2520
ctgtactaca gggagggcaa gttcgtgacc cagaagatcc acaggcagat cgaggacgac 2580
ttcaagaaga tcgacttctc caacaccccg gaggtgaacc tgatcccgtc cggcgtgaag 2640
ctgaagaact tccagttcga gtga 2664
<210> 22
<211> 2766
<212> DNA
<213> 人工序列
<220>
<223> Cas12h.2编码核酸序列
<400> 22
atggccacca ggtccttcat caggaccggc aacctgaagg ccaagaacac cgccgaggag 60
gtgatgcagt ggtacgccga cctgcagtcc gactacaggt ccttcctgaa cctgttcttc 120
ggctggatgg ccatcggcta cggcaccaac gccgaggacg aggtgttcta cacctccaag 180
gaggagtccg agaggctgag gtccctgacc atcggcgacg ccaagaagga gcagctggcc 240
gtgtccttca tcgagctgct gctgaagggc ggcgagaacg cctcctcctg ctacaacgtg 300
ttctacagga actacaagtc cctgggcaag gccaagctga cccagaagaa gaacgacttc 360
ctgtccgccc tgccgctgct ggacgagaac aagatcaagg agtacttcaa gaccgacgag 420
cagctgtccc agatctgcat cgaggagtgg ctggagtacg gcgtgaagaa cctgccgctg 480
ccggagatct gggccgaggt gtccccgagg ctggcctcca tcgagaggtc cctgggcgtg 540
gacctgaggc tggccttcgg cctgtcctgc atcaggtcca gggactgcaa ctactgcagg 600
atcctgatcg agatggtggg cagggacctg aggtccatct tcgagaagta caacaaccac 660
ctgctggaga ccgagaagat caagctgtcc atgaacgaca agcagggccc ggtgtacgac 720
tccatctgct gcttcgccgc cgagctggag tccaagaact ccggcctgac caagtacgtg 780
ctgaccaagg gcatcgacca cgtgaagaag ggcaccggcg agaagaccga catcaggctg 840
gccgtgaagg agctgaagaa gaacaagtac aggatcctga tcgagtcctc ctactccgag 900
atcatgtccg cctactcctg ctggaggacc aagaagcagc tggagaagag gaagctgtac 960
ccgtgcttcg acccgaacag gaacgactac aaggtgccgg tgggccaggg ctccctgggc 1020
aacttcaccg tgtccgtgga ggactccggc gacgtgctga tcgagatcgt gggcgtgggc 1080
gtgatcaggt gcgccgcctc ctgctacttc tccggcatcg tgttcgacga gatcaggaac 1140
aagaacggca ggaccggcta ctccctgaac ttctgccaca agtccatctc caagggcaag 1200
aaggccgtga aggccgcctc ccacaccggc gacaagatct ccggcgtgct gaaggagatc 1260
ggcctgagga acaccgactc cggcttcttc gtgtccctgc cgtactccat ccaccacgac 1320
gagaagaact tcaagatcgc cgagttcttc atgtccgcct gcccgaagaa ggagaacgtg 1380
gagaacctgc cggacaagat cgtggtgggc gccatcgacc tgaacgtgtc caacccggtg 1440
gccgccgtga aggccgtggt gtacagggac gacaagtccg gccagctgaa cgccctggac 1500
tacggctccg gcaacctgat caagaagccg ttcatgctgg tggccaacgg cccgaggatc 1560
aagaacctga tcgagatcag ggacgacgcc aggagggtga tcggcgccat cagggagttc 1620
aaggtgtcca acgccgtgaa ggagcacgtg ggcgaggaca ccagggactt cctgatcctg 1680
tgcggcgaca ccaagtcctc ctccaccagg tacctgatcc agtcctgggt gaagaagatc 1740
aactccaggc tgaggaagat caagttcgag atgaggtccg gcggctacag ggactgcgcc 1800
gacaacatca ggctgatcga ggccatggac cagtgcgcct ccatggccga gtcctacaac 1860
aggatccacc tgaagtccgg cgagaagctg gtgaaggtgg ccaagttcga caagtccagg 1920
gccaacttca ggaacttcgt gctgaggcag ctggcctcca agatcgccaa cgagatgaag 1980
gactgcaacg tggtgttcgg cgaggacctg gacttcatct tcgactccga caagaacaac 2040
aacgccctgc tgaggctgtt ctccgccgcc accctgctga agtacatcat cgaggccctg 2100
gagaagatcg gcgtgggctt cgtgaaggtg gccaagaacg gcacctccca gtccgacccg 2160
gtgacctcca acccgggctg gagggacgac aagaacaagt ccaggctgta cgtggtgagg 2220
gacaagcagc tgggctggat cgactccgac ctggccgcca ccatgaacat cctgatccag 2280
ggcctgaacc actccgtgtg cccgtacaag ttctacgtga aggagtacga gaacaagccg 2340
aactccaccc aggactccat caacgccatc aagaagccgg aggaggccat cggcaagagg 2400
atcaagaggt tcttcaacct gaagtacggc tcctccgtgc cgaagttcgt gtccgacgac 2460
aggggcaggg tgaccttcgc caagaagatc gactccaccc agaccaggct gatcaaccag 2520
ttcgtgtacg cccactcctc ctgcatcgtg acctgcgagc tgcacaacga gatggtgaac 2580
aagatcaagc agctggccgt ggagaagccg aactgccagg agttcgacgt gacctgcgac 2640
ccggacggca ggtacaacaa cttcgccctg ccggaggtgc acgactcctc caaggacgtg 2700
ggcgccaagg ccctgaccac caaggacgtg gacttcaaga ccatcctgaa ggaccacacc 2760
gcctga 2766
<210> 23
<211> 3894
<212> DNA
<213> 人工序列
<220>
<223> Cas12w.1编码核酸序列
<400> 23
atgggcaaga acgagaacaa gtaccagctg tccaagaccc tgaggttcgg cctgaccctg 60
aaggagaaga tctccaacaa cgagaagacc ccgtaccagt cccactccca gttcagggac 120
ctgatcatcc tgtccgagaa caggatcagg gagggcatct ccaccccgca gaacagggac 180
ctgccgtcct tcatccacag gatccagaac tgcaccgact tcatcaacga cttcatccac 240
gactggtgga tgatcctgat gcacaccggc cagatcgagc tggacaagga ctactacaag 300
tccctgacca agaaggtggg cttcgtgggc ttctggtaca aggagaacaa gaagaagggc 360
ggcaagacca agcagccgca ggccaggaac atcccgatgg gcgagctgag gcacctgtgc 420
ccgcagaaca ccaaggagtg cgccacctac atcaccgact actggaagga cctgctgatc 480
accgccacca acaagctgta cgagtcctcc gagcagcaga agaagttcat caaggccatg 540
gagcagaaca ggaccgacaa caagccgaac gagatcgacc tgaagaagtc cttcctgtcc 600
ctggtgtccg tgaccatgga gctgctgaac ccgatcctga acggccagat cctgttcaac 660
aagatggaca ggctggacat gtccaagaag tccgacaacg acttcatcga cttcgtgaac 720
gaccacgaga ccgtgaggga gctgaacaac gacatcgagg agatcatcgc cgacttcaag 780
gagaacggca acaacgtgaa ctactgcaag gccaccctga acccggacac cgccctgaag 840
cagcacaaca acaacatccc gaacgacatc gccaccgacc tggaggagct gatgatggac 900
tccatcgtgg gcaactacga cgacgtgaac tccttcatgg acaactacgt gtccaacctg 960
tccgccaagg acaagatcaa gaagatcaag gactccaaca tctccctgat ctacagggcc 1020
atcctgttca agtacaagat gatcccggcc aacgtgagga gggacatcgc ccagggcatg 1080
gccaagaagc tgaacaagga cgaggagaac atctactcct tcctgtgcga gttcggcacc 1140
ctgaggaccc cgcagaagga ctacgccgac ctgaaggaca aggactcctt caacctggac 1200
aactacccgc tgaaggtggc cttcgacttc gcctgggagg gcctggccaa ggcctggtac 1260
cacgaccagt ccgacttccc gatcgacccg tgcagggact tcctgcagga gaacttcgac 1320
gtgaacctgg aggaggacca ggaggacgag tacttcctgc tgtacgccga cctgatcgag 1380
ctgaacgccc tgctgtccac cctggacaag ggcaacccgg ccgacccgga ctccatcaag 1440
aacgaggccc tggagatggt ggagtacatc aactggaact ccctggacaa gaagaacggc 1500
aactactaca agaagatcat caagaacagg ctgaagtcct ccaagggcaa cgagacctac 1560
gagaggatca agaaggagat ctccatgtcc aggggcaggc tgaagaacaa gatcgagaag 1620
tacgacgacc tgacctccca gtacaagagg atcgccatgg acctgggcaa gaagttcgcc 1680
tccctgaggg acaagatcat cgccgccaac gaggacaaca aggtgaccca ctacgccatg 1740
atcctggagg actccaactg cgacaagtac ctgctgctgc agaaggtgtc caacaacatc 1800
taccactgca tgtcctacga ctcctccgac ccgaaggcct actacgtgga ctccatcacc 1860
tcctccgcca tcgccaagat gatcaggaag gagaccaacc cgtccaagat cagggagtac 1920
gccgagctgg aggagaagga gagggagagg aggaacgtgg acgactggtg caggttcatc 1980
tccaagaagg agtacgacag gaggtaccag ctgaacatca acaacggcct gtccttcgag 2040
gccctgaaga aggagatcga ctccaagtcc tacatcctgg tgaagaagaa catctccgtg 2100
gactccatca gggagctggt ggagaacgag ggctgcctgc tgttcccgat cgtgaacaag 2160
gacctgacca aggagaggaa gaccaccgag gacaaccagt tcaccaagga ctggaacatg 2220
atcttctccg gctccgagac caactggagg ctgaccccgg agttcagggt gacctacagg 2280
aacccggtgc cgggctaccc gaacgacaag ttcggctcca agaggtactc caggttccag 2340
atgaacgccc acttcgtgtg cgacttcatc ccgtcctcca actcctacac ctccaacagg 2400
gagcagatcg ccatcttcaa ggacgagggc gagcagaaga agagggtgga ggagttcaac 2460
aggaccctgt ccaacatcaa ccagaagttc tacgtgatcg gcatcgacag gggccagaag 2520
gagctggcca ccctgtgcgt ggtggaccag gacaagaaga tccacggcga cttcaagatc 2580
tacaccagga agttcaactc cgagaggaag cagtgggagc actactccct ggagggcgag 2640
aagggcacca ggaacatcct ggacctgtcc aacctgaggg tggagaccac catcatcatc 2700
gacggcaagc cggagaggag gcaggtgctg gtggacctgt ccgaggtgct ggtgaaggac 2760
aaggagggca actacaccaa gccgaacaag atgcagatca agatgcagca gatggcctac 2820
gtgaggaagc tgcagttcca gatgcaggcc aacccgaccg aggtgctgga gtggtacgag 2880
cagaacccga ccgaggagct gatcatcaag aacctggtgg acaaggagaa cggcgagaag 2940
ggcctgatct ccttctacgg caccgccctg gtggagctgg accagaccct gccggtgtcc 3000
aagatcaagg agatgctgga ggagttcaag atcctgaagc agagggagtc caagaaggag 3060
aacgtgcaga aggagctgaa caacctgacc cagctggagg ccgtggactc cctgaaggcc 3120
ggcatcgtgg ccaacatggt gggcgtgatc tcctacatcc tgaagaccct ggactacaac 3180
gcctacatct ccctggagga cctgtccacc gtgcagtcct ccaccgagtt cgcctccggc 3240
atctccggcg ccatcaccaa gatgtccagg gaggagggca ggaggatcga cgtggagaag 3300
tacgccggcc tgggcctgta caacttcttc gagatgcagc tgctgaggaa gctgcacagg 3360
atccagaccg acaacggcaa catcctgcac ctggtgccgg ccttcagggc ccagaagaac 3420
tacgaccaca tcatggtggg caaggagaag atcaagaacc agttcggcat cgtgttcttc 3480
gtggacgccg ccgccacctc catcaagtgc ccgaggtgcg gcgccgtgaa cgaggacaag 3540
ttcaacccgg acaagcagaa gtacccggac gccgagaagg gcccgaagct gaggaacagg 3600
aaggagcagt ccggcaagaa ggtgtgggtg accagggaca aggaggacga cgacaggatc 3660
aagtgctact gctgcggctt cgacaccaag gagaagaacg agggcaaccc gttcatgtac 3720
atcaagtccg gcgacgacaa cgccgcctac ctgatctccg acctgggcgt ggagtcctac 3780
aggaaggcct acgagctggc cgccaccgtg gtggaggaca ggaagaagac cctgaccaac 3840
aacctgaacc agtccaacta caagatcagg ttcctgtggc acaccatgta ctga 3894
<210> 24
<211> 3774
<212> DNA
<213> 人工序列
<220>
<223> Cas12w.2编码核酸序列
<400> 24
atggacgccg acaagaccac caaggccatc aacgagtacc agacccagaa gaccatcagg 60
ttcggcctga ccgccaccaa ccagaacctg tactccgagg agatcatgaa gctgctgaac 120
atctccgagg agaggatcat caaggagaag gtgaaggtga acaacgacac cgacaagacc 180
aaccagctga ggggctgcct ggtgcagatc aagaagtacc tgaagacctg ggagaacatc 240
tacgcccaga tcgacttcct ggccatcacc aaggactact acaaggtgat ctccaagaag 300
gccaggttcg acttcgacaa gggcaacggc tccgagatca agctgtcctc cctgcagtcc 360
acccacaaca agaagaagag gtaccagtac atcatcgact tctggaagga gaacctgagg 420
aagaccgaga acctgtacag gaagtccgac gacctgctga agatcttcga ggaggccaag 480
aaccagaaca gggacgacaa gaagctgaac aaggtggagc tgaggaagac cttcctgaac 540
ctgttcaccc tggtgaacga gtccctgaag ccgctgatcg agggcaacct gttcatcgtg 600
aacgacgaca agatcgacga gaagaactcc aagcacaact acgtgttcta cttcatctcc 660
aagaccgagg agaggaggct gctgtacgac aacatctgca ccctgcagga ctacttcaag 720
aacaacggcg gctacgtgcc gttcggcagg gtgaccctga acaagtggac cgccctgcag 780
aagttcaaca acagggacat cgagatcaac aggatcatca aggagctgaa gatcaacaac 840
atctccaccc agaagaccga ctacaagtac aacgacttca ccgagaactt caaggagaag 900
aaggacgaga acggcaaggt ggtgaagaac tccgccggca acatcatctg ggacctgaag 960
gccaacgcca agtccgtgat cgagatctgc cagttcttca agtacaagaa ggtgccgatc 1020
aacgccaggc tgaacctggc caagaggctg atcaaggaca acaagctgaa gaaggagcag 1080
gagaacacct tcctgtccga gttcggcgtg ctgaagaccc cggccttcga ctacgccagg 1140
gacaaggaga acttcaacct gaccaactac ccgctgaagg tggccttcga ctacgcctgg 1200
gagaactgcg ccaaggacaa gtacgagaag atcccgttcc cgaaggagca gtgcgagagg 1260
tacctgcaga ccgccttcga gatcgacgcc accaaggacg agaacaagaa gctgatcgac 1320
acccacctga acaagtacgc cgacctgctg cagttcaaga tcctgctgga gaggttcaag 1380
gccgagttcc acaagaccaa cgaggagacc aacaagaaca acatccagaa gctgaggaac 1440
gtgttctccg gcctggacta ccacggcgac aacaggctga acaagaacca gatccagaag 1500
gccatcgagg cctggttcga caacaaggag cagaacatcg gcaagaagaa ggagaacgag 1560
aagctgctga ccgagaacga gaagaacaac ttctccctgt ccatgcagat catcggccag 1620
gagaggggcg gcctgaagaa cggcatcccg aagtacaagg agctgaccga gatgttcaag 1680
gtgtgcgcct ccaagttcgg caagcagttc gccgacctga gggactactt caacgaggcc 1740
tacgaggtgg acaagatcaa gtacagggcc tggatcatcg aggacgacaa gaagaacagg 1800
ttcgtgctgt tcgtgaacaa ggagaaggcc ttcgacctga cctccgagga gggcgacctg 1860
tggttctacg aggtgaagtc cctgacctcc aagtccctgg tgaagttcat caagaacagg 1920
ggcgcctacc cggacttcca cgacgtgaag aactccttcc actactcctc catcaagaag 1980
gactggcaga actacaagaa cgacccggag ttcctggaca agctgaagga gtgcctgaag 2040
aactccaaga tcgccaagga ccagaagtgg gccaagttct gctgggactt caagcagtgc 2100
gacacctacg agaagctgga gaaggaggtg gacaggaagg gctacaagct ggagggctgc 2160
aagtccgagc cgaagaccat ctccctgacc cagctgaccg actgggtgga gaacaaggac 2220
tgcttcctgc tgccgatcgt gaaccaggac atcaacaagg gcgacaagag gaccaagaac 2280
cagaaccagt tcaccaagga ctggttcgac atcttcgaga acaagaagag gctgcacccg 2340
gagttcaaca tcttctacag gttcccgacc aaggactacc cgaacaccaa gttcaagaac 2400
ggcaccgaga agaccaagag gtactccagg ttccagatgc tggcctactt cggctgcgag 2460
gtgatcccgt ccggcaacca cctgtccaag aaggagcaga tcgccatctt caacaacgac 2520
aagaagcaga aggaggaggt ggagaagtac aacaagtcca tctcctccga ctgcgactac 2580
gtgatcggca tcgacagggg catcaagcag ctggccaccc tgtgcgtgct ggacaagaac 2640
ggcgtgatcc agggcgactt ccagatcttc accaggacct tcaacaagca gaccaagcag 2700
tgggagcaca aggagctgga gcagaggaac atcctggacc tgtccaacct gagggtggag 2760
accaccatca ccggcaagaa ggtgctggtg gacctgtcca agatcaagga cgacgagggc 2820
aactacacca acctgaagca gaccatcaag ctgaagcagc tggcctacat cagggagctg 2880
cagtacgcca tgcagaccag gccggacgac ctgctggact tcgtgaagtc catcaactcc 2940
gccaacgaca tcaccgccga gaacatcaag cacttcatct ccccgtacaa ggagggcaag 3000
aactacgacg acctgccgaa ggtggagatg ttcaacctgc tgaaggagtg gggcaacgcc 3060
gacgagaacg gcaagaggaa gatcgccgag ctggacccgg ccgacaacct gaagtccggc 3120
atcgtggcca acatggtggg cgtggtggcc ttcctgtgcg agaactacaa ctacaaggtg 3180
aggatcgccc tggaggacct gaccagggcc tacggcatcc agaaggacgc cctgaacggc 3240
accgccatct accagaacga cgaggacttc aaggagcagg agaacaggag gctggccggc 3300
gtgggcacca tgcagttctt cgaggtgcag ctgctgagga agctgttcaa gatccaggtg 3360
gacaagaacc tgcacctgat cccggccttc aggtccgtgg acaactacga gaagatcgtg 3420
aggagggaca agcagaactc cggcgacgag ttcgtgaact acccgttcgg catcgtgtgc 3480
ttcgtggacc cgaagtacac ctcccagcag tgcccgtact gcaacaacac ccacaagcac 3540
aagaagaacg acaccgagac cggcaagaag gccttctaca ggaacaaggg cgagaacaag 3600
aactccctgc tgtgcgagaa gtgcggcgtg tccaccatcg agggcgagga gaccctgtcc 3660
tccaagaacg acaacaagaa gcagttcaac atccactaca tcaccgacgg cgaccagaac 3720
ggcgcctacc acatcgccaa caaggtggtg atcaacttcc agaaggactc ctga 3774
<210> 25
<211> 3036
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.1编码核酸序列
<400> 25
atggactacc agcagtacga gttcaccagg accatcaggt tcaacctgtc cggcgacgac 60
aagagggccc tgatgctgga cctgctggac gacacccagg agggcatgct ggccgccttc 120
caggagacct acaagaacct gctgttcgcc ttccaggagg ccatcctgag ggccgacggc 180
tccggcaacc tgagggtggg caggctggag atcaagaagt cctggctgag gcagtacgcc 240
agggagtact tctacgccct gtccgaggac gagaggaggt gcaagaacaa gttccaggcc 300
aagctgttcg acagggtgct gtccgactgg ctggagagga acaacgagct gctgcagagg 360
ctgaacaaca tcctgtccct gccgcaggag tccaagaccg gcgcctccga cctgtccctg 420
ctggtgaggc agctgaaggg cgccgagtac ttctacttca tcagggactt cacccagtcc 480
ggcatcatca acgacaagga ctccgacgag cacatcaaga acctggccgg catcgtggag 540
aagttcgaga ccctgctgga caaggtgctg ttcctgaccg ccccgaactc ctcccagggc 600
gtggagacca ccagggcctc cttcaactac tacaccgtga acaagatctc caagaacttc 660
gacgagaaca tcaagaaggc caacggcagg ctgtgctcct cctaccagaa ctccatgaac 720
gaggagctgc tgaggaaggt gggcttcctg aagtacctga aggacgagta cagggccgag 780
ctgcagaacg tgtccctgaa ggacctgtac gaggccctga agaagttcaa gtcccagcag 840
aagaccgcct tcatccaggc cgtgcagaag aacaagtccg agaaggagct gatgagggag 900
ttcccgctgt tcaacggcaa gcagccggac accctgcaga agttcatcct ggagaccgac 960
aagatcaaga ggggcgccta cttccagaag tggggcttcg acaactacat ctccttctgc 1020
aacaagatct tcaagccggt ggccatggag accggcacca ggaaggccaa gatcagggcc 1080
ctggagcagg agaagatcga ggccaggctg ctgcagtact gggcccacat cctggtgaag 1140
gacggcaagt acttcctgct gctgatcccg aaggagaaga tgggcgaggc caaggtgttc 1200
ttcgccaggc tgtccgacca ggagggcggc gagtacaccc tgtacgcctt caactccctg 1260
accctgaggg ccctgaagaa gctgatcagg aggaacctgg gcaaggagca ggtgaggctg 1320
tccgccggcg acgccgacgc catcgccctg tgccaggagg tgctgagggg caggtaccac 1380
cagctgaagg acctggacct gtccggcttc gagaaggaga tcgccgagat cgccaacacc 1440
cagtacgaga acgaggagga gttcaggatc gccctggagc aggtggccta ctacctgtcc 1500
gagaggaaga tgaacgagga gtccatcgag tacctgaaga agaacctggg cgccatcctg 1560
ctggagatct cctcctacga cctggagagg aacatcaccg gcgagtccaa ggagcacacc 1620
aggctgtggt ccgacttctg gaacccgaac aacaagaagg agtgcttctc caccaggctg 1680
aacccggagc tgaggatctt ctacaggccg ccgagggagc agaaggaccc gaagaagcag 1740
aagaacaggt tctccaagga ccacctggcc gtggccttca ccatcgccca gaacgccgcc 1800
aggaagagga tggagacctc cttcgccgag gagaaggacc tggtggagca ggtgaagaag 1860
ttcaacgagg aggtggtggg caagttcatc gacgagaagt ccgacaacct gtactactac 1920
ggcatcgaca ggggccagca ggagctggcc accctgtgcg tggtgaggtt ctccaaggag 1980
cactacgagg ccatgctgga ggacaacttc atcaagaagt tctccaagcc gatcccggcc 2040
cagatcaccg cctacaggat caaggacgag cacatgtcct acaggaagaa catcaccagg 2100
gacctgaagg gcaacgagac cgaggagatc ctgttcaaga acccgtccca cttcatcgac 2160
gaggtggaga acttcgagga ggtgtccacc ccgtgcatcg acctgaccac cgccaagctg 2220
atcaagggca agatcatcct gaacggcgac atccagacct acctggccct gaagaaggcc 2280
aacggcaaga ggcagctgtt cgagaagttc gccaagatcg acgactccgc caagatcgag 2340
ttcgacgact ccgagggcag gttccaggtg aagtccaagg ccaccgagag ggaggagtac 2400
cagttcctgc cgtactacgg cccggagcag gagaacatct ccccgaggga ggacatgagg 2460
agggagctgc aggcctacct ggacaagctg aggtcctccg agtccttcga ggaggacatc 2520
tccatcgaga agatcaacca cctgagggac gccatcacct ccaacatggt gggcatcatc 2580
gccttcctgt tcaccgagta cccgggcatc atcaacctgg agaacctgca ctccagggag 2640
aacatcgaga agaactggag gaagaacaac gaggacatct ccaggaggct ggagtggggc 2700
ctgtacaaga agttccagaa gatcggcctg gtgccgccga ggctgaggca gaccgtgctg 2760
ctgagggaga acgagaccga gaggcaggag aagctgaacc agttcggcat catccacttc 2820
atcccgaccg agaagacctc cgccaggtgc ccgtactgcg gcgagaacac cccgatgaag 2880
cagaggaacg aggacaagtt caagctgcac gcctacatct gcaggtccaa cgaggagaac 2940
tgcggcttcg acaccaggga gccgaagtcc ccgctggagt tcatcaagaa ctccgacgac 3000
gtggccgcct acaacatcgc caagaagagg ctgtga 3036
<210> 26
<211> 3204
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.2编码核酸序列
<400> 26
atgaagaacg gcatcaacct gttcaagacc aagaccacca agaccaaggg cgtggacatg 60
gagaagtacc agatcaccaa gaccatcagg ttcaagctgc tgccggacaa cgcccacgag 120
atcgtggaga aggtgaagtc cctgaagacc tccaacgtgg acgagctgat ggacgaggtg 180
aagaacgtgc acctgaaggg cctggagctg ctgttcgccc tgaagaagta cttctacttc 240
gacggcaacc agtgcaagtc cttcaagtcc accctggaga tcaaggccag gtggctgagg 300
ctgtacaccc cggaccagta ctacctgaag aagtcctcca agaactccta ccagctgaag 360
tccctgtcct acttcaagga cgtgttcaac gactggctgt tcaactggga ggagtccgtg 420
tccgagctgg ccatcatcta cgagaagtac aagatctgcc agcaccagag ggactccagg 480
gccgacatcg ccctgctgat caagaagctg tccatgaagg agtacttccc gttcatctcc 540
gacctgatcg actgcgtgaa cgacaagaac tccaacaaga ccttcctgat gaagctgtcc 600
gaggagctgt ccgtgctgct ggagaagtgc aactccaggg ccctgccgta ccagtccaac 660
ggcatcgtgg tgggcaaggc ctccctgaac tactacaccg tgtccaagtc cgagaagatg 720
ctgcagaacg agtacgagga cgtgtgccag tccctggaca agaactacga catcaccgag 780
atgaaggtga tcctgtacaa ggagaagctg gacaacctga acttcaagga cgtgaccatc 840
gccaacgcct acaacctgct gaaggagaac aaggccctgc agaagaggct gttctccgag 900
tacgtgtccc agggcaaggt gctgtccctg atcaagaccg agctgccgct gttctccaac 960
atcaacgaca acgacttcga gaagtacaag gagtggtcca acgagatcaa gaagctggcc 1020
gacaagaaga acaccttctg caagaagacc cagcaggaca agatcaagga catccagaac 1080
aagatctccg agctgaagaa gaagaggggc gccctgttcc agtacaagtt cacctccttc 1140
cagaagcact gcgacaacta caagaaggtg gccgtgcagt acggcaagct gaaggccagg 1200
aagaaggcca tcgagaagga cgagatcgag gccaacctgc tgaggtactg gtccgtgatc 1260
ctggagcagg aggacaagca ctccctggtg ctgatcccga agaacaacgc caaggacgcc 1320
aagcagtaca tcgagaccat caacaccaag ggcggcaagt acatcatcca ccacctggac 1380
tccctgaccc tgagggccct gaacaagctg tgcttcaacg ccgtggacat cgagaagggc 1440
cagatggtga gggagaacac cttctaccag ggcatcaagg aggagttcga gaggaacaag 1500
atcaactgcg acaaccaggg cgtgctgaag atccagggcc tgtactcctt caagaccgag 1560
ggcggccaga tcaacgagaa ggaggccgtg gagttcttca aggaggtgct gaagtccaac 1620
tacgccaggg aggtgctgaa cctgccgtac gacctggagt ccaacatctt ccagaaggag 1680
tacaccaacc tggaccagtt caggcaggac ctggagaagt gctgctacgc cctgcactcc 1740
aagatcggca aggacgacct ggacgagttc accaggaggt tcgaggccca ggtgttcgac 1800
atcacctcca tcgacctgaa gtccaagaag gagaagacca agaccaccgg cgagatgaag 1860
aagcacaccc agctgtggct ggagttctgg aagggcgcca tcgagcagaa cttcgccacc 1920
agggtgaacc cggagctgtc catcttctgg agggccccga agtcctccag ggagaagaag 1980
tacggcaagg gctccgacct gtacgacccg aacaagaaca acaggtacct gtacgagcag 2040
tacaccctgg ccctgaccat caccgagaac gccggctccc acttcaagga catcgccttc 2100
aaggacacct ccaagatcaa ggaggccatc aaggagttca acatgtccct gtcccagtcc 2160
aagtactgct tcggcatcga caggggcaac gccgagctgg tgtccctgtg cctgatcaag 2220
aacgagaagg acttcccgtt cgagaagttc ccggtgtaca ggctgaggga cctgacctac 2280
cagggcgact tcaaggacaa gcacgaccag atgaggtacg gcgtggccat caagaacatc 2340
tcctacttca tcgaccagga ggacctgttc gagaagaaca acctgtccgc catcgacatg 2400
accaccgcca agctgatcaa gaacaagatc gtgctgaacg gcgacgtgct gacctacctg 2460
aagctgaagg aggagaccgc caagcacaag ctgacccagt tcttccaggg ctcctccatc 2520
aacaagaact ccagggtgta cttcgacgag gacgagaacg tgttcaagat caccaccaac 2580
aggaaccaca acccggagga gatcatctac ttctacaggg gcgagtacgg cgccatcaag 2640
aacaagaacg acctggagga catcctgaac gagtacctgt gcaagatgga gaccggcgag 2700
tccgagatcg tgctgctgaa cagggtgaac cacctgaggg acgccatctc cgccaacatc 2760
gtgggcatcc tgtcctacct gatcgacctg ttcccggaga ccatcgtggc cctggagaac 2820
ctggccaagg gcaccatcga caggcacgtg tcccagtcct acgagaacat caccaggagg 2880
ttcgagtggg ccctgtacag gaagctgctg aacaagcagc tggccccgcc ggagctgaag 2940
gagaacatcc tgctgaggga gggcgacgac aagatcgacc agttcggcat catccacttc 3000
gtggaggaga agaacacctc caaggactgc ccgaactgca ggaagaccac ccagcagacc 3060
aacgacaaca agttcaagga gaagaagttc gtgtgcaagt cctgcggctt cgacacctcc 3120
aaggacagga agggcatgga ctccctgaac tccccggaca ccgtggccgc ctacaacgtg 3180
gccaggaaga agttcgagtc ctga 3204
<210> 27
<211> 3276
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.3编码核酸序列
<400> 27
atggccggca ccccgtacac cggccacgtg gcctgcaagt actgcaagat cacctcctgg 60
gccacctacg acaggatcaa gatcaacaag atcaacatga accagtcctt catcaacggc 120
cagaacttct acgagctgag gaagaccatc aggttcgtgc tggacccgaa gaccctgaag 180
aggccgtaca ccccgtcctc cgacgaggtg aacctggagg agcagctgaa caacttcatc 240
gagaagtacc agcagggcat caacgacttc aagtacatcg tgtacttcgg cccgaagacc 300
gccgagacca aggagctgaa caagaagatc tccatcaagc actcctggct gaggaactac 360
accaagtccg agttctactc catcaaggac aagctgatcc agctggacta caacggcaac 420
aaggcctcca tcggcaactc caacctgaag ttcctgaacg agtacttcga gaactggatc 480
tccgagaacc aggagtgcgc cgacgccctg aagaactgca tcaacgcccc ggccgagaag 540
cagaagagga agtccgaggc cgcccactgg gtgaggaagc tgaccaagag gtccaacttc 600
gagtgcatct tcgagctgtt caacggcaac atcgaccaca agaactccaa cgacgacatc 660
gagaagatca agcactgcct gaacgagtgc aagaccctgc tgacctccct ggagaagatg 720
ctgctgccgt cccagtccct gggcatggag atcgagaggg cctccctgaa ctactacacc 780
atcaacaaga agccgaagaa ctacgacgag gacatcgccc agaaggcctc cgccctgaac 840
gaggcctacc agttcaaggc cgacgacaag gccttcctga acagggtggg cttctccgac 900
gacggcgtgc cgatcaacga gctgaaggag gccatgaaga agttcaaggc cgaccagaag 960
tccaagttct acgagttcgt gaaccagaag aagtcctact ccgacctgaa gaagaacgac 1020
gacctgaagc tgctgaacga catctccgag gaggacttca acaagttcaa ggagacccag 1080
gacaagatga ccaggggcaa gcacttccag ttctccttcc cgaactacaa gaagtccgag 1140
aagaacttct gcgacctgta caagaacgtg gccgtggcct tcggcaagat cagggccgac 1200
atcaaggccc tggagaagga gaggatggac gccgagaagc tgcagtgctg ggccgtgatc 1260
ctggagaagg acaaccagag gtacgtggtg accatcccga gggacgccaa caacaacctg 1320
accaacacca agcagtacat cgacaacctg cagaacgagg agaacgacca gtggatcctg 1380
tacgccttcg agtccctgac cctgaggtcc ctggacaagc tgtgcttcgg cctggacaag 1440
aacaccttca tcccggccat caccggcgag ctgtaccaga agaacaactc cttcttcgag 1500
aagggcctgc tgaagaggaa ggaccagttc tcccagaacg gcaccgacct ggccgccttc 1560
tacaagaccg tgctggagct ggactccacc aagaagatgc tgggcatcaa caagtacgcc 1620
gacttcaagg ccttcatctc caaggagtac accgccctgg aggacttcga gaagaccctg 1680
aaggagacct gctacttcaa gaagagggtg ttcatctccg aggacaccaa gaacaagctg 1740
atcaacgact accagggcaa cctgtacaag atcacctcct acgacctgga gaaggacgac 1800
tccgaggccc tgggcaccct gatcaacaag aagcagttca acagggcctc cccggagatc 1860
cacaccaaga cctggctgga cttctggacc gccgacaacg agaccgacaa gtacccgatc 1920
aggctgaacc cggagttcaa gatctccttc gtggagaagc aggacaagga cctgaacatg 1980
aggaacctgg gcctgctgaa caagaacagg aggctgaagt cccagttcct gctgtccacc 2040
accatcaccc tgctggccca cgagaagaac gccgacctgc acttcaagaa gaccgacgag 2100
atccagacct tcatcaactc ctacaaccag gagttcaaca agaagatcaa gccgttcgac 2160
atctactact acggcctgga caggggccag aaggagctgc tgaccctggg cctgttcaag 2220
ttctccgaga acgagaaggt gtccttcacc aagcaggacg gcaccgtggg cgagtactcc 2280
aagccgaagt tcatcccgct ggacgtgtac cagatcaggg agggccagta cctgaccaag 2340
aacaagaagg gcaggctggc ctacaagtcc atcgaccagt tcatcgacga cgagaaggtg 2400
atcgagaagc tgccggtgaa ctcctgcctg gacctgtcct gcgccaagct ggtgaagggc 2460
aagatcatcc agaacggcga cgtggccacc tacctggagc tgaagagggt gtccgccctg 2520
aggaagatct acgagaacac caccaggggc cagttcaaga ccgacaggat cggcttcaac 2580
aaggacaagg gctgcctgtt cctggacatc gagaacaggg gcaagctgga gaacaacaac 2640
ctgtacttct acgacaacag gttcgccgag atcctgtccc tggactccat catcaaggag 2700
ctgcaggact actacaacga ggtgaagaac aagcagaaca tcgagttcat ctccatcgac 2760
aagatcaacc acctgaggga cgccctgtgc gccaacgccg tgggcatcct ggcccacctg 2820
cagaagaccc acttcggcgt gatcgtgttc gagggcctgg acgccaggca caagaacaag 2880
gagaccaccg agttcgccgg caacctggcc tccaggatcg agaggaagat cctgcagaag 2940
ctggagaccc tgtccctgat cccgccgcag cacaggcaga tcatcgacct gcagaactcc 3000
aagcagatca agcagaccgg cgccgtgctg tacatcgagg agaagggcac ctccgccaac 3060
tgcccgcact gcgagaccgc caacccggac aagtccgaga agtggctggc ccacaactac 3120
aagtgcaaga actccaactg caacttcgac gcctccgaga tctccaagag gaaggacctg 3180
atcggcctgg acaactccga ctccgtggcc acctacaaca tcgccaagag gggcctgctg 3240
gagatgaacc agaagatcga gcagtccaag gtgtga 3276
<210> 28
<211> 3174
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.4编码核酸序列
<400> 28
atggagatcc aggagctgaa gaacctgtac gaggtgaaga agaccgtgag gttcgagctg 60
aagccgtcca agaagaagat cttcgagggc ggcgacgtga tcaagctgca gaaggacttc 120
gagaaggtgc agaagttctt cctggacatc ttcgtgtaca agaacgagca caccaagctg 180
gagttcaaga agaagaggga gatcaagtac acctggctga ggaccaacac caagaacgag 240
ttctacaact ggaggggcaa gtccgacacc ggcaagaact acgccctgaa caagatcggc 300
ttcctggccg aggagatcct gaggtggctg aacgagtggc aggagctgac caagtccctg 360
aaggacctga cccagaggga ggagcacaag caggagagga agtccgacat cgccttcgtg 420
ctgaggaact tcctgaagag gcagaacctg ccgttcatca aggacttctt caacgccgtg 480
atcgacatcc agggcaagca gggcaaggag tccgacgaca agatcaggaa gttcagggag 540
gagatcaagg agatcgagaa gaacctgaac gcctgctcca gggagtacct gccgacccag 600
tccaacggcg tgctgctgta caaggcctcc ttctcctact acaccctgaa caagaccccg 660
aaggagtacg aggacctgaa gaaggagaag gagtccgagc tgtcctccgt gctgctgaag 720
gagatctaca ggaggaagag gttcaacagg accaccaacc agaaggacac cctgttcgag 780
tgcacctccg actggctggt gaagatcaag ctgggcaagg acatctacga gtggaccctg 840
gacgaggcct accagaagat gaagatctgg aaggccaacc agaagtccaa cttcatcgag 900
gccgtggccg gcgacaagct gacccaccag aacttcagga agcagttccc gctgttcgac 960
gcctccgacg aggacttcga gaccttctac aggctgacca aggccctgga caagaacccg 1020
gagaacgcca agaagatcgc ccagaagagg ggcaagttct tcaacgcccc gaacgagacc 1080
gtgcagacca agaactacca cgagctgtgc gagctgtaca agaggatcgc cgtgaagagg 1140
ggcaagatca tcgccgagat caagggcatc gagaacgagg aggtgcagtc ccagctgctg 1200
acccactggg ccgtgatcgc cgaggagagg gacaagaagt tcatcgtgct gatcccgagg 1260
aagaacggcg gcaagctgga gaaccacaag aacgcccacg ccttcctgca ggagaaggac 1320
aggaaggagc cgaacgacat caaggtgtac cacttcaagt ccctgaccct gaggtccctg 1380
gagaagctgt gcttcaagga ggccaagaac accttcgccc cggagatcaa gaaggagacc 1440
aacccgaaga tctggttccc gacctacaag caggagtgga actccacccc ggagaggctg 1500
atcaagttct acaagcaggt gctgcagtcc aactacgccc agacctacct ggacctggtg 1560
gacttcggca acctgaacac cttcctggag acccacttca ccaccctgga ggagttcgag 1620
tccgacctgg agaagacctg ctacaccaag gtgccggtgt acttcgccaa gaaggagctg 1680
gagaccttcg ccgacgagtt cgaggccgag gtgttcgaga tcaccaccag gtccatctcc 1740
accgagtcca agaggaagga gaacgcccac gccgagatct ggagggactt ctggtccagg 1800
gagaacgagg aggagaacca catcaccagg ctgaacccgg aggtgtccgt gctgtacagg 1860
gacgagatca aggagaagtc caacacctcc aggaagaaca ggaagtccaa cgccaacaac 1920
aggttctccg acccgaggtt caccctggcc accaccatca ccctgaacgc cgacaagaag 1980
aagtccaacc tggccttcaa gaccgtggag gacatcaaca tccacatcga caacttcaac 2040
aagaagttct ccaagaactt ctccggcgag tgggtgtacg gcatcgacag gggcctgaag 2100
gagctggcca ccctgaacgt ggtgaagttc tccgacgtga agaacgtgtt cggcgtgtcc 2160
cagccgaagg agttcgccaa gatcccgatc tacaagctga gggacgagaa ggccatcctg 2220
aaggacgaga acggcctgtc cctgaagaac gccaagggcg aggccaggaa ggtgatcgac 2280
aacatctccg acgtgctgga ggagggcaag gagccggact ccaccctgtt cgagaagagg 2340
gaggtgtcct ccatcgacct gaccagggcc aagctgatca agggccacat catctccaac 2400
ggcgaccaga agacctacct gaagctgaag gagacctccg ccaagaggag gatcttcgag 2460
ctgttctcca ccgccaagat cgacaagtcc tcccagttcc acgtgaggaa gaccatcgag 2520
ctgtccggca ccaagatcta ctggctgtgc gagtggcaga ggcaggactc ctggaggacc 2580
gagaaggtgt ccctgaggaa caccctgaag ggctacctgc agaacctgga cctgaagaac 2640
aggttcgaga acatcgagac catcgagaag atcaaccacc tgagggacgc catcaccgcc 2700
aacatggtgg gcatcctgtc ccacctgcag aacaagctgg agatgcaggg cgtgatcgcc 2760
ctggagaacc tggacaccgt gagggagcag tccaacaaga agatgatcga cgagcacttc 2820
gagcagtcca acgagcacgt gtccaggagg ctggagtggg ccctgtactg caagttcgcc 2880
aacaccggcg aggtgccgcc gcagatcaag gagtccatct tcctgaggga cgagttcaag 2940
gtgtgccaga tcggcatcct gaacttcatc gacgtgaagg gcacctcctc caactgcccg 3000
aactgcgacc aggagtccag gaagaccggc tcccacttca tctgcaactt ccagaacaac 3060
tgcatcttct cctccaagga gaacaggaac ctgctggagc agaacctgca caactccgac 3120
gacgtggccg ccttcaacat cgccaagagg ggcctggaga tcgtgaaggt gtga 3174
<210> 29
<211> 3477
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.5编码核酸序列
<400> 29
atggagaact tcaagaacct gtacgaggtg aggaagaccg tgaggttcga gctgaagccg 60
tccaggaaga agaccttcgc cggcggcgac atcttcgagc tgcagaagga cttcgaggag 120
gtgcagaagt tcttcctgga catcttcgtg ttcgccatcg agcaggagaa gctgtaccag 180
gaggaggagg aggagggcaa gctgtccagg tacaccaaga tcgagttcaa gaagaagagg 240
gagatcaagt acacctggct gaggatctac accaagaacg agttctacga ctggaacggc 300
aagaacgaca aggagaagaa ctacgccctg tccaagatcg acttcctgga gaaggagatc 360
ctgaggtggt tcaacgagtg gcaggagctg accgtgaacc tgaagaacct gacccagacc 420
aaggagcacg agaaggagag gaagtccgac atcgccttcg tgctgaggaa cttcctgaag 480
aggcagaact tcccgttcat caaggacttc ttcaacgccg tgatcgacat ccaggagaag 540
cagggcaacg agtccgacga gaagatcagg aagttcaggg aggagctgag ggagatgaag 600
aagaacctga acacctgcgc caaggagtac ctgtcctccc agtccaaggg cgtgctgctg 660
cacaaggcct ccttcaacta ctacaccctg aacaagaccc cgaaggagta cgagaacctg 720
aagctgcaga aggagctgga gatcgacaac atcctgccga agaagatctg caagagggtg 780
aggtggaaca aggagaagaa gcaggaggac atcctgttcg agtgcaactc cgactggctg 840
gtggagatca agctgggcta cgacatccag aagtggaccc tggacgaggc ctaccagaag 900
atgaagacct ggaaggccga ccagaagtcc gacttcaacg agaagatcgg caacttcatc 960
gaccagtacc tgaagaaggg cttcatcgag gacctgatga acgagaacga gaagaagaac 1020
gccgaggcca tcctgaggga gttctccgtg ttcaagccga tcgagaactt ctacttctac 1080
gacttcctgg agaggaccaa ggagatcaag atcctgtcca accagaagaa caacatcctg 1140
cagaagtaca acaagaacgc caagtacttc gagaagatca tcacctacaa gatcaaggac 1200
aaggaggacc tgaccgagga cgagaaggag taccaggagc tggagaagtc catcgagaag 1260
aaggccaagg agaggggcaa gttcttcaac gccccgaagg agaaggtgca gacccagcac 1320
tacttcgagc tgtgcgagct gtacaagagg atcgccatga agaggggcaa gatcatcgcc 1380
gagatcaagg gcatcgagaa cgaggaggtg cagtcccagc tgctgaccca ctgggccctg 1440
atcgccgagg agggcgagaa gaagtccgtg gtgttcatcc cgaggaagaa cggcgaggag 1500
ctggagaacc acaagaaggc ccacgagttc ctgcagaagc aggagaagaa ggagttcggc 1560
gacatcaagt cctaccactt caagtccctg accctgaggg ccctggagaa gctgtgcttc 1620
aaggagaccg agaacacctt caccccggag atcaagaagg agaccaaccc gaaggtgtgg 1680
ttcccgaagt acaagcagga gtggaacgac gagccgcaga agctgatcaa cttctacaag 1740
caggtgctgc agtccaagta ctcccagaag tacctggacc tggtggcctt cggcgacctg 1800
aagtccttcc tggagacctc cttcgacgac ctgcagatct tcgagtccgg cctggagaag 1860
acctgctaca tcaaggtgcc gatctacttc tccaaggagg gcttcgagac cttcaccaac 1920
aggttcgacg ccgaggtgtt cgagatcacc accaggtcca tctcctccga gtccaagagg 1980
aaggagaacg cccacgccga gatctggaag gacttctggt ccaaggagaa cgaggagaag 2040
aaccacatca ccaggctgaa cccggaggtg tccgtgttct acagggacga gatcgagaag 2100
aagtccaacg ccctgagggg caacaacaag tccaacatca acaacaggtt ctccgcctcc 2160
aggttcaccc tggtgaccac catcaccatc agggccaccc acaagaagtc caacctggcc 2220
ttcaagaccg aggaggacat caagtcccac atcgacaagt tcaacgaggc cttccagaac 2280
ttctccggcg agtgggtgta cggcatcgac aggggcctga aggagctggc caccctgaac 2340
gtggtgaagt tctccgacga gaagaacgag ttcggcgtga tcaagccgaa ggagttcgcc 2400
aagatcccgg tgtacaagct gaaggacgag aaggccatcc tgaaggacga gaacggcaag 2460
gacctgaaga acgccaaggg cgaggccagg aaggtgatcg acaacatctc cgaggtgctg 2520
gaggagaaga aggagccgga ctccaacctg ttcgagaagc agggcgtgct gtcccagggc 2580
atctcctgca tcgacctgac ccaggccaag ctgatcaagg gccacatcat cctgaacggc 2640
gaccagaaga cctacctgaa gctgaaggag atctccgcca agaggaggat cttcgagctg 2700
ttctccacct ccaagatcga caagaactcc gagctgaggg tggagaagac caccatctcc 2760
atcaactccg aggacggcaa gagggacttc tactggctga ccaagaacca gatcgtgaac 2820
tccgagacca agaaggagat ccagaaggag cagcaggaga agctggacaa cctgaaggtg 2880
atcttcatcg actacctgga gggcctgtgc gtgaagaaca agttcgagga catcgagacc 2940
atcgagaaga tcaaccacct gagggacgcc atcaccgcca acatggtggg catcctgttc 3000
cacctgcaga aggagttcaa gggcatcatc gccctggaga acctggacac cgtgagggag 3060
cagtccaaca agaagatgat cgacgagcac ttcgagcagt ccaacgagga catctccagg 3120
aggctggagt gggccctgta caggaagttc gccaacatgg gcgaggtgcc gtcccagatc 3180
aaggagtcca tcttcctgag ggacgagttc aaggtgtacc agatgggcct gctgaagttc 3240
gtggaggtgt ccggcacctc ctccaactgc ccgaactgcg acaaggaggt gggcaagacc 3300
aactcccact tcgtgtgcaa gggcgagaac aactgcggct tctcctccaa ggagaacagg 3360
aacctgctgg agcagaacct gaacaactcc gacgaggtgg ccgcctacaa catcgccaag 3420
aggggcctga agctgatcaa ccagaagtgg aacaacacct ccaagtccca gaactga 3477
<210> 30
<211> 3195
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.6编码核酸序列
<400> 30
atggagaagt acaagatcac caagaccatc aggttcaagc tgctgccgga caagatccag 60
gacatctcca ggcaggtggc cgtgctgcag aactccacca acgccgagaa gaagaacaac 120
ctgctgaggc tggtgcagag gggccaggag ctgccgaagc tgctgaacga gtacatcagg 180
tactccgaca accacaagct gaagtccaac gtgaccgtgc acttcaggtg gctgaggctg 240
ttcaccaagg acctgttcta caactggaag aaggacaaca ccgagaagaa gatcaagatc 300
tccgacgtgg tgtacctgtc ccacgtgttc gaggccttcc tgaaggagtg ggagtccacc 360
atcgagaggg tgaacgccga ctgcaacaag ccggaggagt ccaagaccag ggacgccgag 420
atcgccctgt ccatcaggaa gctgggcatc aagcaccagc tgccgttcat caagggcttc 480
gtggacaact ccaacgacaa gaactccgag gacaccaagt ccaagctgac cgccctgctg 540
tccgagttcg aggccgtgct gaagatctgc gagcagaact acctgccgtc ccagtcctcc 600
ggcatcgcca tcgccaaggc ctccttcaac tactacacca tcaacaagaa gcagaaggac 660
ttcgaggccg agatcgtggc cctgaagaag cagctgcacg ccaggtacgg caacaagaag 720
tacgaccagc tgctgaggga gctgaacctg atcccgctga aggagctgcc gctgaaggag 780
ctgccgctga tcgagttcta ctccgagatc aagaagagga agtccaccaa gaagtccgag 840
ttcctggagg ccgtgtccaa cggcctggtg ttcgacgacc tgaagtccaa gttcccgctg 900
ttccagaccg agtccaacaa gtacgacgag tacctgaagc tgtccaacaa gatcacccag 960
aagtccaccg ccaagtccct gctgtccaag gactccccgg aggcccagaa gctgcagacc 1020
gagatcacca agctgaagaa gaacaggggc gagtacttca agaaggcctt cggcaagtac 1080
gtgcagctgt gcgagctgta caaggagatc gccggcaaga ggggcaagct gaagggccag 1140
atcaagggca tcgagaacga gaggatcgac tcccagaggc tgcagtactg ggccctggtg 1200
ctggaggaca acctgaagca ctccctgatc ctgatcccga aggagaagac caacgagctg 1260
tacaggaagg tgtggggcgc caaggacgac ggcgcctcct cctcctcctc ctccaccctg 1320
tactacttcg agtccatgac ctacagggcc ctgaggaagc tgtgcttcgg catcaacggc 1380
aacaccttcc tgccggagat ccagaaggag ctgccgcagt acaaccagaa ggagttcggc 1440
gagttctgct tccacaagtc caacgacgac aaggagatcg acgagccgaa gctgatctcc 1500
ttctaccagt ccgtgctgaa gaccgacttc gtgaagaaca ccctggccct gccgcagtcc 1560
gtgttcaacg aggtggccat ccagtccttc gagaccaggc aggacttcca gatcgccctg 1620
gagaagtgct gctacgccaa gaagcagatc atctccgagt ccctgaagaa ggagatcctg 1680
gagaactaca acacccagat cttcaagatc acctccctgg acctgcagag gtccgagcag 1740
aagaacctga agggccacac caggatctgg aacaggttct ggaccaagca gaacgaggag 1800
atcaactaca acctgaggct gaacccggag atcgccatcg tgtggaggaa ggccaagaag 1860
accaggatcg agaagtacgg cgagaggtcc gtgctgtacg agccggagaa gaggaacagg 1920
tacctgcacg agcagtacac cctgtgcacc accgtgaccg acaacgccct gaacaacgag 1980
atcaccttcg ccttcgagga caccaagaag aagggcaccg agatcgtgaa gtacaacgag 2040
aagatcaacc agaccctgaa gaaggagttc aacaagaacc agctgtggtt ctacggcatc 2100
gacgccggcg agatcgagct ggccaccctg gccctgatga acaaggacaa ggagccgcag 2160
ctgttcaccg tgtacgagct gaagaagctg gacttcttca agcacggcta catctacaac 2220
aaggagaggg agctggtgat cagggagaag ccgtacaagg ccatccagaa cctgtcctac 2280
ttcctgaacg aggagctgta cgagaagacc ttcagggacg gcaagttcaa cgagacctac 2340
aacgagctgt tcaaggagaa gcacgtgtcc gccatcgacc tgaccaccgc caaggtgatc 2400
aacggcaaga tcatcctgaa cggcgacatg atcaccttcc tgaacctgag gatcctgcac 2460
gcccagagga agatctacga ggagctgatc gagaacccgc acgccgagct gaaggagaag 2520
gactacaagc tgtacttcga gatcgagggc aaggacaagg acatctacat ctccaggctg 2580
gacttcgagt acatcaagcc gtaccaggag atctccaact acctgttcgc ctacttcgcc 2640
tcccagcaga tcaacgaggc cagggaggag gagcagatca accagaccaa gagggccctg 2700
gccggcaaca tgatcggcgt gatctactac ctgtaccaga agtacagggg catcatctcc 2760
atcgaggacc tgaagcagac caaggtggag tccgacagga acaagttcga gggcaacatc 2820
gagaggccgc tggagtgggc cctgtacagg aagttccagc aggagggcta cgtgccgccg 2880
atctccgagc tgatcaagct gagggagctg gagaagttcc cgctgaagga cgtgaagcag 2940
ccgaagtacg agaacatcca gcagttcggc atcatcaagt tcgtgtcccc ggaggagacc 3000
tccaccacct gcccgaagtg cctgaggagg ttcaaggact acgacaagaa caagcaggag 3060
ggcttctgca agtgccagtg cggcttcgac accaggaacg acctgaaggg cttcgagggc 3120
ctgaacgacc cggacaaggt ggccgccttc aacatcgcca agaggggctt cgaggacctg 3180
cagaagtaca agtga 3195
<210> 31
<211> 3321
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.7编码核酸序列
<400> 31
atgctgatcc agttcaagaa ccactactcc tacaacaagt ccatcaggtt caagctggag 60
cacaagaacg gcaagctgcc gaagctggag tccgacaacg tggacctgaa caagctggtg 120
gacatcggca actccctgaa ggacatcttc gaggagctgg tgtacaccaa gaacaactac 180
aacaagctga actccctggt gtccatcaag aagcagtggc tgaagatcta cttcaagaac 240
gagttctact ccaacggcaa gatccagaac tactccctgt ccaacttctc ctacctgccg 300
aacaagctga tcgagtggct gaacaactgg cagaacaacc tgaaggccct gatcgagctg 360
accaagcagc aggacttcaa caagaccaag aagtccgaga tcgcctacat cctgtccctg 420
ttcaacggca agtactcctt ctccttcgtg aaggacttct ccacctgcat caaccacaag 480
aactcccagg agcagatcct gaagctgcag ggcgtggtgg agaacttcga gaaggtgctg 540
aacctgtgca tccaggagta cctgccgtcc aagtccgccg gcgtggtgat cgcccagggc 600
tccatgaact actacgccat caacaaggag ccgaagaggt acgacaacat cctggccgac 660
ctgaaccaga agttcgagga gctggacaag gagtacatcg ccatgaagca gtacaagtcc 720
tcccagaagt ccaggctgtt cgagttcatc aggaagggct tctccaagga ccagatcctg 780
tccgagttca agaagaagga gaacaacgag gtgtccttcg tgtacaacaa ccagatcatc 840
atcaggatct acacccagga gctgttcaag gactcctact gcctgggcga ggtgatcaag 900
ctgaccaaga agatcgagga gctgaacgag tccaaggact ccaacaacaa cctgccggag 960
gagaccaaga aggagatcac caagctgaag aaggagatcg gcttctactt catcaggagg 1020
accaggggca agtcccacaa caactacttc aagtcctact acggcttctg caacgacaag 1080
ttcaagaaga aggcccagga gaggggcagg ctgctgacca agatcaaggc catcaggaag 1140
gagaagatcg agtcccagaa cctgaggtac tggtccctga tcctggacga cggcaaggac 1200
aagttcctgt ggctggtgcc gaaggagaac atgcaggagt tcaggaggga gctgtccaag 1260
atccacccgt ccggcgagtc ctccctgttc ctgttccact ccctgaccat gagggccctg 1320
cacaagctgt gcttcgccca ggagtccgac ttcgtgaagg agatgccgaa ggtgctgaag 1380
gaggagcagc tgaactgcga gaaggcctcc aacgacaccg agaccaacaa gaggatcaag 1440
aggaacttcg gcctgaacta catcaagacc aaggacgagc tgaccctgtc cttcctgaag 1500
aagctgatca tctccgagta cgcccacgag aggctggacc tgaaccactt cgacctgtcc 1560
aagctgcagg tggccaccac cctgaacgag ttcgaggagt acctggagga cgcctgctac 1620
tacctggaga agatctccat ctcctcctcc atgatcaagg agctgctgga ggagtacaac 1680
atcctgaact tcaggatcac ctcctacgac ctggagaaga ggaacaagaa cacctaccag 1740
accccggagt ccgacatcaa gaggcacacc aaggagatct ggaacaagtt ctgggagggc 1800
gacaggttca tcaggctgaa cccggagatc aagatcaggt acaggcagaa gaaccagaac 1860
atcgaggact acctgaagga gaagggcttc gacctgacca agatcaagaa caggttcctg 1920
caggagcagt actccgtgtc cttcaccttc gccctgaacg ccggcaagaa gtacccgaag 1980
ctggccttcg tgaagaccga ggagatcctg gagaagatcg aggagttcaa cgacgagttc 2040
aacaagcagt acttcgacaa ctcctacaag tacggcatcg acaggggcaa catcgagctg 2100
gccaccctgt gcatcaccaa gttcaacaag aacgacacct acgagtacaa gggcaagaag 2160
tacctgaagc cgaacttccc gacctcccag gaggacatca agacctacga gctgaagaac 2220
gagtggtaca agaggaccgc catctccaac atcgagacca agccgaagaa caagaagacc 2280
ccgaagagga tcatcgccaa catctcctac ttcatcgaca acgtggagaa cgaggagtgg 2340
ttcaacaaga agacctgcac ctccatcgac ctgaccaccg ccaaggtgat caagggcaag 2400
ctgatcctga acggcgacgt gctgaccttc ctgaagctga agaaggaggc cgccaagagg 2460
atcctgttcg agctggtggc ccagaacaag ctgaccgcca agaacaagga gctgaagtgg 2520
aagtccgacg acggcaacaa ctccgactcc gtgaggctga tctgcgacgt gctggacaac 2580
gagaccaact ccatctactt ctacgaggac tccaagtacg gcaggggctt cgagggcctg 2640
ctgaccaccg acaagaccgc ctactccaag gagggcatca ggatcaacct gcagaactac 2700
ctgaaccacc tgatctccga gaaggagaac aagtccaaca aggcctactc ccacgtgccg 2760
tccatcgaga agatcaacca cctgagggac gccctggtgg ccaacatggt gggcgtgatc 2820
tcctacctgc aggcctacta cccgggcatc gtggtgctgg aggacctgaa ccacaagctg 2880
ctgatcaagc acttcgagga cctgaacatc aacatctcca acaggttcga gcacgccctg 2940
atcgagaagt tccagaccct gggcatggtg ccgccgcaca tcaaggacta cctggagatc 3000
aggtcctcct tcaggatgtc caggaacgac tcctcccagt tcggcgccct gatcttcgtg 3060
tccaaggagg gcacctccaa ggagtgcccg tactgcgaga agaagtggaa ctggggcaag 3120
gagaaggaga tcgagctgaa gttctccaag aagcagtaca tctgcggcaa ggagaactcc 3180
tgcggcttcg acaccaagca catccagaac accttcgagt tcctgtccga gatcaacgac 3240
ccggacaaga tcgccgccta caacatcgcc aagaggggct tcaagtcctt catcaacaag 3300
tcctccatca agaagcagtg a 3321
<210> 32
<211> 3258
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.8编码核酸序列
<400> 32
atggagaagt acaagatcac caagaccatc aggttcaagc tgctgccgga caagatccag 60
gacatctcca ggcaggtggc cgtgctgcag aactccacca acgccgagaa gaagaacaac 120
ctgctgaggc tgatccagag gggccaggag ctgccgaagc tgctgaacga gtacatcagg 180
tactccgaca accacaagct gaagtccaac gtgaccgtgc acttcaggtg gctgaggctg 240
ttcaccaagg acctgttcta caactggaag aaggacaaca ccgagaagaa gatcaagatc 300
tccgacgtgg actacctgtc cagggtgttc gaggacttct tcaacgagtg ggagaccgtg 360
atcgagagga tcaacaccga ctgcaacagg ccggaggagt ccaagaccag ggacgccgag 420
atcgccttct ccatcaagaa gatcgccacc aagcagatgt tcccgttcat caagtccttc 480
gtgtacaact ccaactacaa gaactccgag gagaccaagt ccaagctgac cgccctgctg 540
aacgagttcg agaccgtgct gaagatctgc gagcagaact acctgccgtc ccagtccgcc 600
ggcatcgtga tcgccaaggc ctccttcaac tactacacca tcaacaagaa gcagaaggac 660
tacaagggct acaccgacga catcgagaag atcgagaagg gcatgaactc caagttccac 720
tacgagagga agtacgacca gctgctggag gagctgaacc tgatcgccct gaaggagctg 780
ccgctgatcg agttctactc caagatcaag tcctacaagt ccaccaggaa gatcgagttc 840
tccgaggccg tgtccaaggg cctggccttc gccgacctga agtccaagtt cccgctgttc 900
cagaccgagt ccaacaagta cgccgagttc ctggagctga ccggcaggat cacccagatc 960
tccaccgcca agtccctgct gtccaaggac aacccggagg cccagaagct gagggacgag 1020
atcaagaagc tgaggatcaa caggggcgag tacttcaaga acaacttcca caagtacatc 1080
tccctgtgca acctgtacaa gaagatcgcc gacaagaagg gcaggctgaa gggccaggtg 1140
aagggcatcg agaacgagag gatcgactcc cagaggatcc agcactgggc cctggtgctg 1200
gaggacaacc tgaagcactc cctgatcctg atcccgaagg agaaggtgac cgaggtgtac 1260
aggaaggtga gggcctccaa ggccgactcc acctcctcct cctcctccct gtactacttc 1320
gagtccatga cctacagggc cctgcacaag ctgtgcttcg gcgtgaacgg caacaccttc 1380
ctgccggaga tccagaagga gctgccggag tacaacccga acaagcagtc cgacttcggc 1440
gagttctgct tccacaagtc caacaccgac aaggagatcg acgagccgaa gctgatctcc 1500
ttctaccagt ccgtgctgaa gaccaactac gtgaaggaca acctgaacct gccgcagtcc 1560
gtgttcgacg aggccaccgt gcagaccttc gagaccaggc aggacttcca gatcgccctg 1620
gagaagtgct gctacgccaa gaagaccatc atctccgaga ccctgaagaa ggagatcctg 1680
gaggacaaca acgtgcagat cttccagatc acctccctgg acctgcagag gtccgagcag 1740
aagaacctga aggcccacac caagatctgg aacaggttct ggaccaagca gaacgagacc 1800
gccaactacg acctgaggct gaacccggag accgccatcg tgtggaggaa gccgaagaag 1860
accaggatcg acaagtacgg cgccggcacc tccctgtacg acccgaagaa gaggaacagg 1920
tacctgcacg agcagtacac cctgtgcacc accgtgaccg acaacgccct gaacaacgag 1980
atcaccttcg ccttcgagga caccaagaag aagggcaccg agatcgtgaa gtacaacgag 2040
aagatcaacc agaccctgaa gaaggagttc aacaagaacc agctgtggtt ctacggcatc 2100
gacgccggcg agatcgagct ggccaccctg gccctgatga acaaggacaa ggagccgcag 2160
ctgttcaccg tgtacgagct gaagaagtcc gacttcttca agcacggcta catctacaac 2220
aaggagaggg agctggtgat cagggagaag ccgtacaagg ccatccagaa cctgtcctac 2280
ttcctgaacg aggagctgta cgagaagacc ttcagggacg gcaagttcca ggagaccttc 2340
aacgagctgt tcaaggagaa gcacgtgtcc gccatcgacc tgaccaccgc caaggtgatc 2400
aacggcaaga tcatcctgaa cggcgacatg atcaccttcc tgaacctgag gatcctgcac 2460
gccaagagga agatctacga ggagctgatc atcaacccgc aggccgagct gaaggagaac 2520
gagaaggagt actacctgta cttcgacaag gagggcaccg agaaggtgga gaagatctac 2580
aggtccaggc tggacttcga gcacatcaag ccgtaccagg agatcaggaa cgacctgaac 2640
gcctacttca agaacgtgca gaagaacgag gccaaggtgg aggaccagat caaccagacc 2700
aggagggccc tggtgggcaa catgatcggc gtgatctact acctgtacca gaagtacagg 2760
ggcatcatct ccatcgagga cctgaagcag accaaggtgg agtccgacag gaacaagttc 2820
gagggcaaca tcgagaggcc gctggagtgg gccctgtaca ggaagttcca gcaggagggc 2880
tacgtgccgc cgatctccga gctgatcaag ctgagggagc tggagaagtt cccgctgaag 2940
gacgtgaagc agccgaagta cgagaacatc cagcagttcg gcatcatcaa gttcgtgtcc 3000
ccggaggaga cctccaccac ctgcccgtcc tgcgagaaga aggcctacga gctgcagaag 3060
gagaagaagg gcgaggagaa gccggccgag aacaagaggt acgaggccga caagaaggcc 3120
ggcgtgttct gctgcccgaa gtgcggcttc cacaacagga ccaacccgat gggctacgag 3180
tccctggact ccaacgacaa ggtggccgcc ttcaacatcg ccaagagggg cttcgaggac 3240
ctgcagaagc acaagtga 3258
<210> 33
<211> 3102
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.9编码核酸序列
<400> 33
atggagaact ccaacctgta ccaggtggtg aagaccatca ggttcaagct ggagccggtg 60
ggcaagatgg acaccccgaa gttcggcgac aagaacgccg agtccaaggc caacctgacc 120
ccgttcatcg agctggtgaa gaagaccatg accaacgtga aggccctggt gttctccaag 180
caggacggcg aggacggcga gaagtggagg aagatcctgg aggtgaacta caggttcctg 240
aggtcctacc tgaagaactc cttctacgag aacaggggcg actcccagga gaagtccaag 300
aagcacaaga tctccgacct ggagtacctg cagaaggccc tggagaacct gttcgccgag 360
ttcgacgaga tcctggacgg cctggaggac ttcgagaaga ggaacaccaa gaaccagtac 420
gagaagcaga ggcacgccca ggccggcctg ctgctgaaca ggctgtgcaa gaggtccaac 480
ttcggcttcc tgaaggcctt cgtgggcgcc ctggcccaga ccaacaagcc gttcttcgac 540
gacaagaccg acaagctgaa gaagcagatc gacaagttcg agaccgagct ggagaagcag 600
aaggagttct tcctgccgta ccagtccaac ggcgtgctgt tcgccggcgg ctccttcaac 660
aggtacgcca tcaacaagac cccgaagatg ctggacaagg agctgaggga ggagcagacc 720
aacctgaaga agtccctgtg cgagcacaag atcaagatcg acaccctgaa caccctgggc 780
ctgaagaacg actgcccgtg cacctccctg gacaactcct acaccttcat caaggactac 840
aaggccaagc agaagtccaa gttcatcgag ctggtgcaga agggcgagtt cgacgaggcc 900
aagaaggtga acctgttcga gtgctccgag accgacttcg agaccttcaa gaccaggacc 960
aagcagatcc agaacgagaa ggacaaggac gagaggacca agctgaagca gaagaggggc 1020
gagttcttca agtcccagaa gaggggcaag ttcttcaagt cccagaccca gaactacgag 1080
aacctgtgcg acctgtacaa gaagatcgcc cagaagaggg gccagatcgt ggccaagatc 1140
tgcgccatca agaaggagaa ggagatgtgc gagcaggtga agtactggtg cgtggccctg 1200
gagaagggcg gcgagttcta cctgtacatg ttcctgaggg acgagaacga caacatcaag 1260
aacgcctacg acttcgtgtc caagctgcag acccagaagt ccggcgagac caagctgcac 1320
tacttcgact ccctgaccct gaaggccgtg aggaagctgt gcttcaagga gaccgacggc 1380
tccttcaaga aggccctgaa gaacgtgaag ttcccggagt gcgagcagaa cctggacgag 1440
aaggtgaaga tctccttcta ccagaacgtg ctgaagaacg ccaagaccct gaacctgtcc 1500
aagttcgaga acctgcagtc cgtgaccgag ggcaagttcg agtccctgtc cgagttcgag 1560
gtggccctga acatggtgtg ctacaccaag accgtgtgcg tgtccgagtc cgtggagaag 1620
gagctgaaga agttcaagcc gctggtgttc cacatcacct cccaggacct ggccgccaag 1680
agggagaaga aggcccacac ccagatctgg cacgagttct ggagggagtc caacgagaag 1740
tccaagttcc cgctgaggct gaacccggag ctgaaggtga tgtggaggga ggccaggccg 1800
tccagggtgg agaagtacgc cgagcagtcc gacaagttcg acccgaacaa gaagaacagg 1860
tacctgcacc cgcagttcac cctggccctg aacttcaccc agaacgccca caacgaggcc 1920
atcaacctgg ccttcaagga cgtgcagaac aagggcgagg ccgtgaagaa gttcaacgag 1980
aacttcaagt cctccgagta cgccttcggc atcgacgtgg gcaccaagga cctggccctg 2040
ctgtgcctga tcgacaagaa caagaagccg gtgaacttcg acgtgtacga gatctgcaac 2100
gagaacgaga tctgcaacga gaagctgggc ttcgagaagt tcggcttcta caaggacggc 2160
accaggaggg acgagccgta caagctgatc aagaacccgt cctacttcct gaacgagtcc 2220
ctgtacaaga agaccttcaa cgccaccaag gaggagttcg agaggtcctt ctccgagctg 2280
ttcaagagga agtccgtgtg cgccctggac ctgaccaccg ccaaggtgat ctgcggcaag 2340
atcatcctga acggcgactt ctccacccac ctgaacctga agatcctgaa cgccaagagg 2400
aagatctccg ccaagctgaa gaaggacccg accctgaaga tcgagtacga caacgacgac 2460
aacatcctgt tcggctccaa cgtgatcttc tactacaaca acaagtacga gatcgtgagg 2520
ccgtacgacg agatcaagaa cgagatcttc gagttccacg agaagcagag gctggacgac 2580
gccaggctgg aggacaacat caacaagacc agggccaacc tggtggccaa catggtgggc 2640
gtgatctcct tcctgcacaa ggagttctcc ggcttcgtgg tgctggagaa cctgaagcag 2700
tccgagatcg agggcaacca caggctgaag ttcgagggcg acatcaccag gccgctggag 2760
ctggccctgt acaggaagtt ccagtccaag tgcctgaccc cgccgatctc cgagctgatc 2820
aagctgaggg agggcgagaa gaacgagaac gtggagtccg acctgatcct gcagttcggc 2880
atcatcaagt tcgtggacaa ggacaagacc tccaggctgt gcccggcctg cggcaaggac 2940
gcctacgaga acaacaactc caagtacaag accgacaaga aggacggcgt gttcgagtgc 3000
gccggctgcg gcttcaacaa caagaacaac gccggcgact tcgccgccct ggacaccaac 3060
gacaagatcg ccaccttcaa catcgccaag aggggcctgt ga 3102
<210> 34
<211> 3507
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.10编码核酸序列
<400> 34
atggagacct acaagatcac caagaccatc aggttcaagc tggaggccga cgaggagaac 60
tccatccaca tcaaggagga catcatcaac atcgagacca acgacaacga gttcaccatg 120
gtggacttcg tgtccaacct gggcaactac atcaaggacc tgaagaacta cctgttctac 180
gagaagaagg acggctccct gtccttcaag gacaagatca tcatcaagaa cgagtggctg 240
aggcagtacg ccaagcagga cttcgtggag ctgaagtcca agaagaggat caacctgagg 300
aacaacagga tggagcagat caagatcggc gacatcccga ggctgtcctc caagatcgag 360
gaggccctgg acatcgccaa ggagatctac tccaagctgt ccgacgacgc caccctggag 420
cagcacgaga ggaccaagaa ggcccagatc ggcctgctgc tgaagaggct ggaggccaag 480
aacgtgctgc cgctgctgat ggacctggtg aaggagaccc tggacaagga cgagaccgac 540
gacctgtcca tcaggctgaa gaggcagtcc cagaagatca actcccagct gaagatcgcc 600
atcaggtcct tcctgccgga gcagtccaac ggcctgcaga tcgccaaggc ctccttcaac 660
tactacacca tcaacaagaa gccgatcgac ttcgagaaga agatcgagga cctgaagaag 720
aacctgaacg tgaaggacct ggagaagctg aacgtgtact tcgacaagaa ggagaagaag 780
cagaagaact acctgggcaa gaagatcttc tccctgttcg agaccgacat ccagaaggcc 840
ctgtccaaga accagccgct gtacctgggc gacgccccga tgatcgactc cgcctacgtg 900
tccctgaggc agatcttcaa gaagatcaag tccgagcaga agaagcagtt ctccgagctg 960
atgcagaaca agtgctccta cgacgagctg aagaactcca acctgtacct gctgaacgac 1020
atcggcctgg agcagttcaa cacctacagg gagaagacca aggagctgga ggagctggcc 1080
accaagctgt ccaaccagaa cctgctggag aacgccaagg agaggctgag gtcccagaag 1140
gagaagatcg ccaaggagag gggcaacatc atgaaggaca ggttccagac ctggaagtcc 1200
ttcgccaact tctacaggac cgtgtcccag aagcacggca agatcctggc ccagctgaag 1260
ggcatcgaga aggagcaggc cgagtcccag ctgctgaagt actgggccct gatctgcgag 1320
aaggagaacc agcaccagct gtggctgatc ccgagggaga aggcctggga gtgcaagagg 1380
tggctggaga ccgtgaacga cacctccatc gacaacgaga actccatcaa gctgtactgg 1440
ttcgagtccc tgacctacag gtccctgcag aagctgtgct tcggcttcct ggagaacggc 1500
aacaacgagt tcaaccagaa catcaaggac ctgctgccga aggacaggat cggcaacacc 1560
atcaacggcg agttcgcctt cgagggcgac gaggagagga agatcgagtt ctacaagacc 1620
gtgctgaact ccaagtacgc caagcaggtg ctgaacatcc cgttcaagca ggtggaggag 1680
gagatcatct cccagtcctt cgagaacctg tccgacttcc agatcgccct ggagaagatc 1740
tgctacagga ggttcgccat ctactccaac tacatcatct ccttcgacgc ccagatcttc 1800
gacatcacct ccctggacct gaagaacaac gagaagaaca acctgaacac ccacacccac 1860
atctggaggg acttctggaa ggacgagaac gagaagaaca acttcgacat caggctgaac 1920
ccggagatca ccatctccta caggaccccg aagcagtcca ggatcgagaa gtacggcgag 1980
aagaccaagg agtacgaccc gaacaagaac aacaggtacc tgcacccgca gttcaccctg 2040
atcaccacca tctccgagag gtccaactcc cagaccaaga ccctgtcctt catcgaggac 2100
gaggacttca agaagtccat caacgagttc aacaagaagc tgaagaagga caacatcaag 2160
ttcgccttcg gcatcgacaa cggcgaggtg gagctgtcca ccctgggcgt gtacctgccg 2220
accttcgaga aggagaccca cgaggagaag atctacgagc tgaagcagat caagaagtac 2280
ggcttcgagg tgctgaccat caccgacctg aagtacaagg agaccgacta caacggcaac 2340
gtgaggaaga tcatccagaa cccgtcctac ttcctgaaga aggagaacta catcaggacc 2400
ttctccaagt ccgagcagga gtacgaggag atgttcgcca agctgttcaa gaaggagcac 2460
gtgctgtccc tggacctgac caccgccaag atgatctgcg gccacatcgt gaccaacggc 2520
gacgtgccgg ccctgttcaa cctgtggctg aagcacgccc agaggaacgt gttcgagatg 2580
aacgaccaca ccgtgaagga gaccgccaag accatcaggc tgaggaacaa cgaggagctg 2640
accgacaacg agaaggagaa gttcgccgag ttcatctccg acggcaagaa gttcgccaag 2700
ctgaccaagg agggcaagaa gtccaggtac ctgaagtgga tcttcgagga caggaaggag 2760
aactccttca ccgaggacga gaacaagaag ttcaacgact gccagaagaa gaagggcaag 2820
tacaactccc acatcatctt cgcctccagg ttcgagggcg acgagctgaa gtccgtgacc 2880
ccgatcttcg actgcaggca cgtgttcaag aagaggaagg agttcgagac catcaggccg 2940
atcaaggaga tcgagaacga gatctccagg ttcaacacca acaggacctc ccacaacatc 3000
tccaacgagg agctggacct gaagatcacc gacgccaaga aggccctggt ggccaacgcc 3060
atcggcgtga tcgacttcct gtacaagcag tacaagcaga ggttcaacga cgagggcctg 3120
atcatcaagg agggcttcga cacccagaag gtggaggagg acatcgagaa gttctccggc 3180
aacatctaca ggatcctgga gaggaagctg taccagaagt tccagaacta cggcctggtg 3240
ccgccgatca agaacctgat ggccgtgagg aacgagggca tcaaggacaa gaacgccatc 3300
ctgaggctgg gcaacatcgc cttcatcgac ccgtccggca cctcccagga gtgcccggtg 3360
tgcaaggaga agtccaagga gaagcacacc aacaacttca tctgcgagtg cggcttcaac 3420
tccaccaaca tcatgcactc caacgacggc atcgccggct tcaacatcgc caagaggggc 3480
ttcgagaact tcatcaacga gaagtga 3507
<210> 35
<211> 3564
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.11编码核酸序列
<400> 35
atggagaagt acaagatcac caagaccatc aggttcaggc tggacgccga caacaccgcc 60
atctccgcca tcgtgaagga caccgaggcc ctggaggcca ggggccaggg cttcaagatc 120
aagaagttcg tgaacgccct gggcaggttc ctgtccggcg acggcgtgca gaagtacctg 180
tacgacatgt ccaacgagga gaactgcgtg ttcaagagga acctggtgat caagaacacc 240
tggctgaaga acaacgccaa gcaggagatc gccggcatgg acctgaagag gggcctgatc 300
atcaaggaca tcaagggcct gcaggacaag atcgaggaga tctacgacaa gctgtgggag 360
atctacgaga tcctgtacga gtccgcctac ctgccgctgc aggacctggc caggagggag 420
ggcatcggcc tgctgctgaa gaagctgtcc gtgaagaacg ccctgccgtt catcatctcc 480
ttcgtggagg agtccaacga caagaacgag gccgacgacc tgtccctgag gctgaagaag 540
cagggcaagg agatcctgac ccagctggag atcggcatca acgagtacct gccggcccag 600
tcctccggcc tgccggtggc caaggcctcc ttcaactact acaccatcaa caagaccccg 660
gtggacttcg gcgagaagat ccaggagctg gagaagaggc tgtccgtgga catcaagaag 720
gagatctcct ccttcaccgg cggcatcaag accgccatca agaacaagat cgccggcaag 780
aagatcctgc tgggcgacac cccgatgttc gagtccgaga actccgtgtc cctgaggcag 840
atcctgaaga acatcaagtc cgagcagaag gcccagttca acaagttcat gaccacccag 900
aacaacccgc agctggagga gatgaagacc atgggctggt acctgttcgg cgacatcacc 960
gagggcgagt tcaacgacta caaggagcag accaaggaga tcgagagggt gggcgccaag 1020
atcaaccagt gcggcaacat caaggagaag aaggagctga ggtcccagct gcagaagctg 1080
aagaagaaga ggggcgagct gatctccgag gcccacaaga agggcggcaa cgacaagaac 1140
ttcaagacct acaaggagtt cgccaagttc tacaggaaga tcgcccagag gcacggcaag 1200
atcctggccc agatcaaggg catcgagaag gagaagatcg actccgccat gctgaactac 1260
tgggccgccg tgatcgagct gtccggcagg cacaagctgg tgctgatccc gaagaaggac 1320
gagaacgcca agaagtgcat cgagtggctg gaggacgagt ccaagcacaa gaacggctcc 1380
tgcaagatct tctggttcga gtccttcacc ttcaggtccc tgcagaagct gtgcttcggc 1440
aacctggact ccggcaccaa caccttcaac cagaagatcc agaacctgct gccgtgcgac 1500
gagaggggca acctgatgaa cggcgagttc gccttcaagg gcgacgagca ggagaagatc 1560
aagttctaca agaaggtgct gcagtcccag aaggacatca acctgccgca gaaggaggtg 1620
gtggacaacg tggtgggcag gaagttcgag accatggacg agttcaagat cgccctggag 1680
gagatctgct acatcaggag ggagaggctg tccgccaacg ccgagtccga gctgaagtcc 1740
aagttcaacg cccagatctt cgacatcacc tccctggacc tgaggaaccc ggtgaactgc 1800
gccggcaagc cggaggtgta ccaccacaac gacaagaggc acaccgagat ctggaaggag 1860
ttctggtccc tggacaacga gaggaggaac ttcaacatca ggctgaaccc ggagatcacc 1920
atcacctaca ggaagccgaa ggagtccagg atcctgaagt acggcaaggg caccgagaag 1980
tacaacgccg acatgaagaa caggtacctg tacccgcagt acaccctgct gaccaccatc 2040
tccgagcact gcaacacccc gaccaagatc ctgtccttca tgaccgacaa cgagtacgag 2100
gagtccatca aggccttcaa ctccaagctg aagaaggagg acatcaagtt cgccttcggc 2160
atcgactccg gcgagaccga gctgtccacc ctgggcgtgt acctgccgga gttctccgcc 2220
gagtccaccg agctgaagga catcgagaag tacggcttca acgtgctgac catcaaggac 2280
ctgaactaca ccgagaccga ctacaacggc tccgacaaga agatcgtgaa gaacccgtcc 2340
tacttcgtgg acaagtccct gtacatgagg accttcaaga agaccgagca ggagtacgag 2400
aagatgttcg ccgagcagtt cgaggccaag aagaggctgt ccctggacct gtccgccgcc 2460
aaggtgatct gcggccacat cgtgaccaac ggcggcgtgt ccgagcactt cggcctgtgg 2520
ctgaagcacg cccagaggac catcttctgg atgaacgacc acaccgagaa gaagaccgcc 2580
aagaacatca agctgaagga ctcctccgag ctgacctacg acgagaggga gaagttcgcc 2640
gagcacatct cctccgacga gaagttcaag aagctggacg tggaggagaa gaagaggtac 2700
gtgaggtgga tcttcgagga cagggagacc ctgaacttca ccgaggccga gaacaagaag 2760
ttcggcggct accagaagaa gaagggcgac tacaggctgg gcatcctgtt cgcctcctgc 2820
ttcatcggca aggagctgga gtccgtgacc cagatcctgg actgcaggca catcttcaag 2880
aagagggagg agttctactc cctgaagtcc aaggaggaca tcgaggccga gatcaagagg 2940
tacaacaccg actacaccaa ccacaacatc tccaccgagc agctggacct gaagttcgtg 3000
aacgtgaaga acgccctggt ggccaacgcc gtgggcgtga tcgacctgct gtacaagcag 3060
tacaaggaga ggctgggcgg cgagggcctg atcgccaagg agggcttcga caccaagaag 3120
gtggaggagg acatggagaa gttctccggc aacatctaca ggatcctgga gaggaagctg 3180
taccagaagt tccagaacta cggcctggtg ccgccgatca agaacctgat ggccgtgagg 3240
gccgacaagg tggagatctc cgaggccgag aagtccaaga tcagggagaa ctgcaagatc 3300
tccaagatcg acccggagaa cgagatcatc aagaggaaca agaccctgat cctgaggctg 3360
ggctccatcg ccttcgtgaa cgacgccgac acctcccagg agtgcccggc ctgcggcacc 3420
aagtccaagg agaagcacgt ggacaacttc atctgcggct gcggcttcaa ctccaccggc 3480
atcatccact ccaacgacgg cgtggccggc ttcaacatcg ccaagagggg cttcgtgaac 3540
ctgatggagc acgagctgag gtga 3564
<210> 36
<211> 3588
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.12编码核酸序列
<400> 36
atggagaagt acaagctgac caagaccatc aggttcaagc tgaagccgaa ggacatctcc 60
gccatcaaga gggacgtgga ggccctggag cagcagaagt tcgacctggt gctgttcgtg 120
tacaacctgc acaacttcat cggcaagctg aaggagtacc tgttcttcca gaaggagaag 180
gacgagttcg tgatcaagga caagctgacc atcaagaaga cctggctgaa gcagtacgcc 240
aagcaggaga tcgccggcct ggagctgaac agggagcaga ccctgggcaa catcaagggc 300
gtgtccgcca ggatcgagag ggccgtggac gacgtgaaca agatctacgt ggagctggcc 360
atggaggcca agctgaacga gagggccaag aaggccaaga ccgagcagct gatcaagagg 420
ctggacacca ggaacgccct gccgctgctg gtgtccctga tcgagcagtc ctccgacaag 480
tacgagaccg gcaacctgtc catccagctg aagaggctgg gcaagaggct gcagacccag 540
ctgctgtccg gcatcaagaa gtacctggcc gagcagtcca acggcctgcc gatcgccaag 600
gcctccttca actactacgc catcaacaag aagccggtgg actacatcga caagatcaag 660
cagctgcaga aggacctgga gatcaagaag aacaggaggt ccgaggagag gtacgacaag 720
aagaagagga agaacatcaa gatcttcaac gactccaagc tgtggatcaa gatcaagaag 780
gacatcgaga aggagagggg caacaagacc ctgatcctgg gctacgcccc gatgatcgag 840
ccgggcaact acgtgtacct gaggcagatc ctgaagaaca tcaagctgga gcagaagaac 900
aagttctcca agctgatgca gtccaagtcc ctgaccttcc acgacctgaa caacaacaac 960
cagctgtacc tgttcaagga catcctggag ggcgagttca acaagtacaa gcagaagacc 1020
aacgagatcg agaccaaggc cgagaagagg aaccagtgca acaacgacga gctgaagagg 1080
aagctgaact ccgagctgca gcagctgagg aaggacaggg gctccctgat caacgccgcc 1140
gacggcaggc cgaagggcag gttcaagacc tacaagtact tcgccaactt ctacaggaac 1200
gtggcccaga agcacggcag gatcctgtcc accctgaagg gcatcgagaa ggagatggtg 1260
gagtcccagc tgctgaagta ctggaccatc atcaccgagg agaacaacca gcactccctg 1320
gtgctgatcc cgaaggagag ggccggcgag tacaagaagg acctggagaa ctccatcccg 1380
tccgacccgt cctccaagat caaggtgtac tggttcgagt ccttcaccct gaggtccctg 1440
aggaagctgt gcttcggcta cgtgaacaac aacaccggct ccaacacctt ctacccggag 1500
ctgaagaagt ccgacgagct gaggaagtac cacgacgaga ggggcaactt catcaagggc 1560
gagttctact tcaagggcga cgagcagaag atcatccagt tctacaagga cgtgctgagg 1620
tccaactacg cccagaaggt gctgaagttc ccgaagcagc aggtgaagga cgagctgatc 1680
ggcagggagt tctcctccct ggacgagttc cagatcgccc tggagaagat ctgctaccag 1740
aggcacgtgg tgtgctccca gaaggtggtg gacgccctgt ccaggtacaa cgcccagatc 1800
ttcctgatca cctccctgga cctgggcaac ccggccaact gcgtggacaa gccgaagcag 1860
ttctcccact tcgacaagaa gcacaccagg atctggaagg agttctggtc ctccaagaac 1920
gagaccgcca acttcgacat caggctgaac ccggagatcg tgatcaccta caggcagccg 1980
aagcagtcca ggatcaagaa gtacggcccg gagtccacca ggtacgacga caggaagcac 2040
aacaggtacc tgtacccgca gttcaccctg atcaccacca tctccgagta ctccaacgcc 2100
ccgaccaagg ccctgtcctt cctgaccgac gaggagttca agggcgccgt ggacgagttc 2160
aacaagaagt tcaagaagga gaacatcagg ttctccctgg gcatcgacaa cggcgagacc 2220
gagctgtcca ccctgggcgt gtacctgccg gtgttcaaga aggactccaa cgagaaggtg 2280
gtggccgagc tgaagaaggt gaacaagtac ggcttcaact tcctgaccat caaggacctg 2340
tcccacgtgg agaaggacaa gaacggcagg gtgaggaaga tcatccagaa cccgtcctac 2400
ttcctgtcca aggagcagta catgaggacc ttcggcagga ccgagcagga gtacaacaac 2460
atgttcgccg agcagttcga ggagaaggcc ttcctgtccc tggacctgac caccgccaag 2520
gtgatcaacg gccacatcgt gaccaacggc gacgtgccga ccttcctgaa cctgtggatg 2580
aggcacgccc agagggacat ctgggacatg aacgaccaca ccaaggagaa gaccgccaag 2640
aagatcgtga tcaagaacaa cgacgagctg accgacgccg agaaggtgaa gttcgtggag 2700
tacatctccg acgagaccaa ctacgccaag ctgaacttca acgagaagaa gaggtacgtg 2760
ctgtggatct tcgagaacag gaagaacatc aacttcaccg acgccgagaa gaagaagttc 2820
gagccgtgcc agaagaggaa gggcaacttc tccaaggaca tcctgttcgc cgtgtgctac 2880
atcggctccg agatccactc cgtgaccaac atcttcgacg tgaggaacat cttcaagatg 2940
aggaaggact tctacgtgct gaagtccgag atggagatca agaaggagat cgagtcctac 3000
aacaccaccg ccggcatcca ggagatctcc aacgaggagc tggacctgaa gatcaacagg 3060
ctgaagcagg ccgtggtggc caacgccgtg ggcgtgatcg actacctgta catctactac 3120
aagaagaaga ccggcggcga gggcctgatc atcaaggagg gcttcgacac caagaaggtg 3180
gccaaggccc tggagaagtt ctccggcaac atctacagga tcctggagag gaagctgtac 3240
cagaagttcc agaactacgg cctggtgccg ccgatcaagt ccctgatggc cgtgagggag 3300
gagggcatcg agaacaacaa ggacgccatc ctgaggctgg gcaacgtggg cttcatcgac 3360
ccgaccggca cctcccagca gtgcccggtg tgctccaagg gcaagctgaa ccacaccacc 3420
aagtgctcca agaactgcgg cttcaactcc aagaacatca tgcactccaa cgacggcatc 3480
gccggctaca acatcgccaa gaggggcttc gagaacttca tctcccagaa gaagggctac 3540
gacgtgatca acaacggcac caagtacaac aacctgaagt cccagtga 3588
<210> 37
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12g.1原型同向重复序列
<400> 37
gucuaauaug auaguaaaau auuauauagu uuagac 36
<210> 38
<211> 37
<212> RNA
<213> 人工序列
<220>
<223> Cas12g.2原型同向重复序列
<400> 38
uguguuggua ucuaguaaaa ucuagagccg uugacac 37
<210> 39
<211> 40
<212> RNA
<213> 人工序列
<220>
<223> Cas12h.1原型同向重复序列
<400> 39
augugcugca uggcaacgcu agaugccaug uguucgcaac 40
<210> 40
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12h.2原型同向重复序列
<400> 40
gugcugaugc uauucaauag gauagaauca aggcac 36
<210> 41
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12w.1原型同向重复序列
<400> 41
gucuaaaccg acccaauaau uucuacuguu guagau 36
<210> 42
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12w.2原型同向重复序列
<400> 42
gccuacaaag gcacacaaau uucuacuauu guagau 36
<210> 43
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12j.1原型同向重复序列
<400> 43
cucuaauacc uauacacaau uucuacuuuu guagau 36
<210> 44
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12j.2原型同向重复序列
<400> 44
aucgaacaga cucauaaaau uucuacuguu guagau 36
<210> 45
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12j.3原型同向重复序列
<400> 45
ggcuaauauc ccuaucagau uucuacuuuu guagau 36
<210> 46
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12j.4原型同向重复序列
<400> 46
cucuacaacu gauaaagaau uucuacuuuu guagau 36
<210> 47
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12j.5原型同向重复序列
<400> 47
guuuaauucg uauauuuaau uucuacuuuu guagau 36
<210> 48
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12j.6原型同向重复序列
<400> 48
guccaaagga cggauuaaau uucuacuauu guagau 36
<210> 49
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12j.7原型同向重复序列
<400> 49
gucuagaagc augcucuaau uucuacuguu guagau 36
<210> 50
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12j.8原型同向重复序列
<400> 50
guccaaagga cggauuaaau uucuacuacu guagau 36
<210> 51
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12j.9原型同向重复序列
<400> 51
gucuagacca caaauuuaau uucuacuauu guagau 36
<210> 52
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12j.10原型同向重复序列
<400> 52
guuuaggagu uaaauagaau uucuacuauu guagau 36
<210> 53
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12j.11原型同向重复序列
<400> 53
gucuauaggc ggguuuuaau uucuacuauu guagau 36
<210> 54
<211> 36
<212> RNA
<213> 人工序列
<220>
<223> Cas12j.12原型同向重复序列
<400> 54
gucuaauagu uaaucaaaau uucuacuauu guagau 36
<210> 55
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12g.1原型同向重复序列的编码核酸序列
<400> 55
gtctaatatg atagtaaaat attatatagt ttagac 36
<210> 56
<211> 37
<212> DNA
<213> 人工序列
<220>
<223> Cas12g.2原型同向重复序列的编码核酸序列
<400> 56
tgtgttggta tctagtaaaa tctagagccg ttgacac 37
<210> 57
<211> 40
<212> DNA
<213> 人工序列
<220>
<223> Cas12h.1原型同向重复序列的编码核酸序列
<400> 57
atgtgctgca tggcaacgct agatgccatg tgttcgcaac 40
<210> 58
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12h.2原型同向重复序列的编码核酸序列
<400> 58
gtgctgatgc tattcaatag gatagaatca aggcac 36
<210> 59
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12w.1原型同向重复序列的编码核酸序列
<400> 59
gtctaaaccg acccaataat ttctactgtt gtagat 36
<210> 60
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12w.2原型同向重复序列的编码核酸序列
<400> 60
gcctacaaag gcacacaaat ttctactatt gtagat 36
<210> 61
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.1原型同向重复序列的编码核酸序列
<400> 61
ctctaatacc tatacacaat ttctactttt gtagat 36
<210> 62
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.2原型同向重复序列的编码核酸序列
<400> 62
atcgaacaga ctcataaaat ttctactgtt gtagat 36
<210> 63
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.3原型同向重复序列的编码核酸序列
<400> 63
ggctaatatc cctatcagat ttctactttt gtagat 36
<210> 64
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.4原型同向重复序列的编码核酸序列
<400> 64
ctctacaact gataaagaat ttctactttt gtagat 36
<210> 65
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.5原型同向重复序列的编码核酸序列
<400> 65
gtttaattcg tatatttaat ttctactttt gtagat 36
<210> 66
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.6原型同向重复序列的编码核酸序列
<400> 66
gtccaaagga cggattaaat ttctactatt gtagat 36
<210> 67
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.7原型同向重复序列的编码核酸序列
<400> 67
gtctagaagc atgctctaat ttctactgtt gtagat 36
<210> 68
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.8原型同向重复序列的编码核酸序列
<400> 68
gtccaaagga cggattaaat ttctactact gtagat 36
<210> 69
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.9原型同向重复序列的编码核酸序列
<400> 69
gtctagacca caaatttaat ttctactatt gtagat 36
<210> 70
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.10原型同向重复序列的编码核酸序列
<400> 70
gtttaggagt taaatagaat ttctactatt gtagat 36
<210> 71
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.11原型同向重复序列的编码核酸序列
<400> 71
gtctataggc gggttttaat ttctactatt gtagat 36
<210> 72
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> Cas12j.12原型同向重复序列的编码核酸序列
<400> 72
gtctaatagt taatcaaaat ttctactatt gtagat 36
<210> 73
<211> 11
<212> PRT
<213> 人工序列
<220>
<223> NLS序列
<400> 73
Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val
1 5 10
<210> 74
<211> 893
<212> PRT
<213> 人工序列
<220>
<223> Cas12g.1-NLS融合蛋白的氨基酸序列
<400> 74
Met Leu Tyr Thr Met Asn Val Lys Thr Ile Lys Leu Lys Val Asp Ala
1 5 10 15
Thr Lys Glu Val Glu Ser Arg Leu Thr Lys Met Leu Leu Val His Asn
20 25 30
Asn Ile Gly Arg Glu Ile Ile Asn Phe Leu Ile Leu Cys Ser Gly Asn
35 40 45
Asp Asn Ile Arg Lys Thr Lys Phe Asp Glu Phe Gly Asn Ser Tyr Asp
50 55 60
Glu Phe Cys Asn Leu Lys Leu Asp Gln Phe Asn Leu Tyr Asp Arg Leu
65 70 75 80
Thr Glu Ile His Asp Glu Val Thr Leu Glu Asp Phe Gln Lys Thr Leu
85 90 95
Asn Asp Ile Tyr Asp Leu Val Leu Asn Ser Lys Ser Phe Ser Asn Val
100 105 110
Ser Ser Thr Ile Phe Asn Lys Asn Lys Lys Val Asn Phe Asp Glu Thr
115 120 125
Lys Lys Gly Asp Leu Ser Arg Lys Cys Leu Met Asn Ala Arg Asp Trp
130 135 140
Gly Val Leu Pro Leu Ile Ser Val Asp Asp Asp Ile Val Thr Cys Gly
145 150 155 160
Thr Leu Lys Gly Ile Leu Ser Glu Cys Gln Ser Arg Ile Leu Ser Trp
165 170 175
Asn Glu Cys Asn Leu Ser Thr Lys Glu Thr Tyr Ser Glu Lys Lys Ser
180 185 190
Glu Tyr Gln Ser Ile Leu Asp Asp Ser Met Thr Lys Asp Ala Asp Val
195 200 205
Thr Thr Ala Met Ile Gln Phe Met Asp Asp Val Ser Asn Val Tyr Gly
210 215 220
Ser Asn Asn Glu Asn Gln Leu Lys Trp Phe Asn Asn Arg Phe Leu Thr
225 230 235 240
Tyr Val Arg Asn Lys Ile Arg Pro Phe Leu Leu Thr Asn Ser Pro Ile
245 250 255
Asp Asn Phe Glu Gln Ser Asp Thr Ser Tyr Asn Cys Ser Ile Glu Ile
260 265 270
Val Arg Ile Leu Ser Lys Tyr Glu Ile Leu Trp Lys Asp Glu Val Ser
275 280 285
Val Asn Arg Tyr Lys Lys Thr Cys Asp Asp Gly Ile Asn Ile Glu Lys
290 295 300
Tyr Arg Tyr Leu Val His Ala Lys Ser Asp Phe Leu Arg Tyr Lys Glu
305 310 315 320
Thr Ala Ser Phe Lys Glu Ile His Ala Val Lys Ser Pro Ile Ser Leu
325 330 335
Cys Phe Gly Asn Asn Tyr Gln Pro Phe Ser Leu Ser Asp Val Gly Asp
340 345 350
Arg His Asn Ile Asn Phe Gly Tyr Lys Phe Gly Lys Leu Gly Lys Gln
355 360 365
Arg Lys Glu Cys Ser Phe Asn Leu Asn Tyr Arg Arg Lys Lys Val Lys
370 375 380
Tyr Ala Asn Thr Pro Val Arg Ser Asp Glu Asn Lys Cys Tyr Leu Asp
385 390 395 400
Asn Leu Glu Ile Glu Asp Ala Lys Asn Gly Ser Tyr Lys Leu Ser Tyr
405 410 415
Met Val Asn Lys Lys Tyr Lys Arg Glu Ser Phe Ile Lys Glu Pro Lys
420 425 430
Met Lys Met Tyr Asn Gly Lys Leu Tyr Met Tyr Phe Pro Met Ser Asn
435 440 445
Glu Phe Glu Glu Asp Arg Asp Ser Phe Ala Leu Leu Thr Tyr Phe Ser
450 455 460
Arg Ser Ser Asn Ser Lys Ser Gln Ile Asp Glu Ala Ser Asn Ile Leu
465 470 475 480
Gln Asn Arg Lys Ile Arg Val Cys Gly Val Asp Leu Gly Ile Asn Pro
485 490 495
Thr Phe Ala Leu Ser Val Leu Glu Tyr Ser Asp Asn Lys Ile Thr Asp
500 505 510
Thr Asn Ile Gly Met Lys His Glu Gly Ser Tyr Asn Asn Phe Ser Glu
515 520 525
Ile Arg Lys Gln Ile Asn Asp Val Thr Asp Met Ile Ser Tyr Leu Lys
530 535 540
Ser Lys Tyr Asp Asn Cys Glu Lys Asp Tyr Ser Ser Lys Ile Asp Asp
545 550 555 560
His Ile Lys Ser Arg Leu Asn Glu Glu Ile Ser Asn Phe Cys Asp Leu
565 570 575
Val Ser Tyr Lys Arg Asn Lys Asn Thr Ile Ile Arg Lys Glu Ile Lys
580 585 590
Asn Val Glu Lys Glu Ile Asn Lys Ile Lys Asn Cys Arg Arg His Thr
595 600 605
Leu Lys Lys Asp Leu Thr Glu Asn Phe Gly Trp Val Ser Ala Leu Asn
610 615 620
Glu Phe Ile Ser Leu Lys His Ser Phe Asn Asp Met Gly Glu Ser Phe
625 630 635 640
Asp Ser Lys Thr Asn Pro Ser Tyr Ser Tyr Phe Glu Lys Trp Lys Arg
645 650 655
Tyr Ile Asp Asn Ile Lys Asp Asp Ser Leu Lys Thr Val Ser Arg Glu
660 665 670
Ile Leu Asn Phe Cys Ile Glu Asn Ser Val Asp Phe Ile Ala Leu Glu
675 680 685
Asp Leu Gln Thr Phe Ala Pro Ser Asp Asp Arg Thr Lys Ser His Asn
690 695 700
Lys Leu Thr Gln Leu Trp Cys Phe Gly Lys Leu Lys Lys Cys Leu Glu
705 710 715 720
Asp Ile Ala Ser Met Tyr Gly Ile His Val Tyr Ser Ser Thr Asp Pro
725 730 735
Arg Asn Thr Ser Asp Thr His Phe Glu Ser Lys Asn Phe Gly Tyr Arg
740 745 750
Asp Glu Ser Asn Lys His Asn Leu Trp Val Asn Val Asp Gly Glu Tyr
755 760 765
Thr Val Val Asp Ser Asp Ile Asn Ala Ser Lys Asn Ile Ala Asn Arg
770 775 780
Phe Leu Thr His His Lys Asp Leu Lys Gln Leu Pro Met Ile Gly Asp
785 790 795 800
Gly Thr Leu Phe Lys Ile Asp Ser Ser Ser Lys Arg Asn Lys Ser Phe
805 810 815
Ala Val Lys Leu Asn Ile His Lys Asn Val Tyr Glu Leu Ile Asp Gly
820 825 830
Glu Phe Val Lys Ser Asn Lys Lys Pro Asn Gly Thr Ser Arg Lys Gln
835 840 845
Thr Ala Tyr Ile His Gly Asp Met Phe Ile Asp Ser Ile Ser His Lys
850 855 860
Asn Lys Lys Met Phe Leu Arg Glu Asn Leu Ile Arg Asn Gly Phe Ile
865 870 875 880
Ser Lys Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val
885 890
<210> 75
<211> 946
<212> PRT
<213> 人工序列
<220>
<223> Cas12g.2-NLS融合蛋白的氨基酸序列
<400> 75
Met Asn Lys Thr Asp Thr Gln Asn Asn Glu Gln Ile Asn Lys Pro Thr
1 5 10 15
Gln Leu Leu Asn Asn Lys Asp Ile Glu Leu Thr Val Lys Thr Val Lys
20 25 30
Ser Ala Thr Val Lys Val Asp Asn Asn Ser Lys Lys Glu Leu Phe Gly
35 40 45
Leu Phe Asn Tyr Phe Thr Ser Val Ala Ser Gly Ile Lys Asp Lys Val
50 55 60
Tyr Asn Leu Gln Ser Asp Glu Lys Thr Ala Pro Ile Phe Asn Asp Tyr
65 70 75 80
Val Lys Gln Pro Gln Arg Gly Arg Ser Ala Ala Thr Thr Leu Phe Thr
85 90 95
Lys Leu Asp Ala Glu Lys Thr Tyr Thr Ser Gln His Ser Phe Pro Gly
100 105 110
Lys Trp Arg Asp Ser Gly Ile Phe Pro Leu Tyr Asn Lys Glu Ser Glu
115 120 125
Lys Tyr Asp Leu Ser Thr His Gly Tyr His Tyr Ser Ala Asn Ala Glu
130 135 140
Ile His Thr Gln Leu Asp Ser His Asp Glu Cys Asn Lys Glu Cys Glu
145 150 155 160
Lys Glu Tyr Ala Ala Leu Arg Asp Glu Val Asn Asn Tyr Lys Tyr Glu
165 170 175
Phe Thr Leu Gln Phe Lys Ala Glu Asn Ala Glu Lys Phe Tyr Asn Phe
180 185 190
Val Glu Lys Leu Thr Leu Met Gly Trp Arg Tyr Asp Ala Thr Phe Arg
195 200 205
Ser Phe Phe Glu Leu His Met His Pro Lys Leu Lys Thr Gly Glu Thr
210 215 220
Thr Tyr Arg Ala Thr Tyr Lys Leu Pro Ser Gly Lys Ser Lys Arg Tyr
225 230 235 240
Ser Phe Phe Arg Asp Asp Ile Ala Asp Glu Ile Ala Lys Asn Pro Glu
245 250 255
Phe Trp Pro Met Leu Glu Ser Ser Asn Ala Ile Ser Trp Ile Asn Ser
260 265 270
Asn Asn Leu Leu Ser Arg Lys Lys Asp Lys Ala Asn Tyr Ser Ser Thr
275 280 285
Ser Leu Ile Lys Ser Gln Ile Arg Leu Tyr Leu Gly Asn Asn Gly Val
290 295 300
Pro Phe Thr Ala Arg Glu His Asp Gly Arg Ile Tyr Phe Ser Phe Arg
305 310 315 320
Leu Pro Ala Ile Asn Gly Glu Lys Gly Arg Met Val Glu Ile Pro Cys
325 330 335
Ser Tyr Lys Lys Val Phe Asn Gly Lys Ala Arg Lys Ser Cys Tyr Leu
340 345 350
Gly Gly Leu Thr Ile Glu Lys Thr Asp Ala Gly Lys His Ile Phe Lys
355 360 365
Tyr Ser Val Asn Asn Lys Lys Pro Gln Val Ala Glu Leu Asn Glu Cys
370 375 380
Phe Leu Arg Leu Val Val Arg Asn Arg Glu Tyr Phe Asn Asn Val Val
385 390 395 400
Ala Gly Lys Ile Thr Asp Ile Asn Thr Asp His Phe Asp Phe Tyr Val
405 410 415
Asp Leu Pro Leu Asn Val Lys Glu Asp Pro Ile His Asp Leu Ser Ser
420 425 430
Thr Glu Val Phe Gly Lys Asn Gly Leu Arg Ser Tyr Tyr Ser Ser Ala
435 440 445
Tyr Pro Glu Ile Lys Asn Leu Gly Ser Gln Ile Glu Thr Gly Lys Asn
450 455 460
Leu Thr Cys Pro Ile Thr Lys Thr His Asn Ile Met Gly Ile Asp Leu
465 470 475 480
Gly Gln Arg Asn Pro Phe Ala Tyr Cys Ile Lys Asp Asn Thr Gly Lys
485 490 495
Leu Ile Ala Gln Gly His Met Asp Gly Ser Lys Asn Glu Thr Tyr Lys
500 505 510
Lys Tyr Ile Asn Phe Gly Lys Glu Ser Thr Ser Val Ser His Leu Ile
515 520 525
Lys Glu Thr Arg Ser Tyr Leu His Gly Asp Pro Glu Ala Ile Ser Lys
530 535 540
Glu Leu Tyr Asn Glu Val Ala Gly Phe Cys Asn Asn Pro Val Ser Tyr
545 550 555 560
Glu Glu Tyr Leu Lys Tyr Leu Asp Ser Lys Lys Phe Leu Ile Asn Lys
565 570 575
Glu Asp Leu Ser Lys Asn Ala Met His Leu Leu Arg Gln Lys Asp His
580 585 590
Asn Trp Ile Gly Arg Asp Trp Leu Trp Tyr Ile Ser Lys Gln Tyr Lys
595 600 605
Lys His Asn Glu Asn Arg Met Gln Asp Ala Asp Trp Arg Gln Thr Leu
610 615 620
Tyr Trp Ile Asp Ser Leu Tyr Arg Tyr Ile Asp Val Met Lys Ser Phe
625 630 635 640
His Asn Phe Gly Ser Phe Tyr Asp Lys Asn Leu Lys Lys Lys Val Asn
645 650 655
Gly Thr Val Val Gly Phe Cys Lys Thr Val His Asp Gln Ile Asn Asn
660 665 670
Asn Asn Asp Asp Met Phe Lys Lys Phe Thr Asn Glu Leu Met Ser Val
675 680 685
Ile Arg Glu His Lys Val Ser Val Val Ala Leu Glu Lys Met Asp Ser
690 695 700
Met Leu Gly Asp Lys Ser Arg His Thr Phe Glu Asn Arg Asn Tyr Asn
705 710 715 720
Leu Trp Pro Val Gly Gln Leu Lys Thr Phe Met Glu Gly Lys Leu Glu
725 730 735
Ser Phe Asn Val Ala Leu Ile Glu Ile Asp Glu Arg Asn Thr Ser Gln
740 745 750
Val Cys Lys Glu Asn Trp Ser Tyr Arg Glu Ala Asp Asp Leu Tyr Tyr
755 760 765
Val Thr Asp Gly Glu Ser His Lys Val His Ala Asp Glu Asn Ala Ala
770 775 780
Asn Asn Ile Val Asp Arg Cys Ile Ser Arg His Thr Asn Met Phe Ser
785 790 795 800
Leu His Met Val Asn Pro Lys Asp Asp Tyr Tyr Val Pro Thr Cys Ile
805 810 815
Trp Asp Thr Thr Glu Glu Ser Gly Lys Arg Val Arg Gly Phe Leu Thr
820 825 830
Lys Leu Tyr Lys Asn Ser Asp Val Val Phe Thr Lys Lys Gly Asp Lys
835 840 845
Leu Val Lys Ser Lys Thr Ser Val Lys Glu Leu Lys Lys Leu Val Gly
850 855 860
Lys Thr Lys Glu Lys Arg Gly Gln Tyr Trp Tyr Arg Phe Glu Gly Lys
865 870 875 880
Ser Trp Ile Asn Glu Ala Asp Arg Asp Thr Ile Ile Leu Asn Ala Lys
885 890 895
Lys Ile Ser Arg Glu Arg Asp Asn Gly Glu Gln Ser Thr Asp Thr Arg
900 905 910
Ser Gln Asn Val Thr Val Ser Val Leu Asp Val Cys Glu Thr Ala Glu
915 920 925
Lys Lys Lys Leu Val Leu Val Ser Arg Ala Asp Pro Lys Lys Lys Arg
930 935 940
Lys Val
945
<210> 76
<211> 898
<212> PRT
<213> 人工序列
<220>
<223> Cas12h.1-NLS融合蛋白的氨基酸序列
<400> 76
Met Ala Leu Ile Gln Arg Ala Gly Val Leu Lys Thr Lys Ser Asp Phe
1 5 10 15
Pro Lys Val Ile Lys Asp Trp His Asp Ser Leu Leu Ala Asp Tyr Arg
20 25 30
Lys Phe Phe Pro Ile Ile Phe Ser Trp Cys Pro Glu Tyr Gly Tyr Thr
35 40 45
Thr Ile Gln Asp Asn Lys Pro Val Phe Val Ser Pro Glu Glu Arg Met
50 55 60
Glu Ser Ile Arg Lys Glu Ala Lys Glu His Leu Asn Glu Val Leu Ala
65 70 75 80
Phe Gly Lys Met Ile Gly Ser Lys Gly Val Gly Gly Ser Ser Ser Tyr
85 90 95
Ala Ile Phe Tyr Lys His His Lys Asn Asn Glu Asn Gly Ala Tyr Thr
100 105 110
Pro Ser Arg Ala Lys Phe Met Lys Glu Gly Ile His Asn Arg Arg Val
115 120 125
Glu Leu Val Asp Val Leu Met Leu Asn Ala Ile Pro Asp Glu Glu Trp
130 135 140
Val Lys Ile Ala Gln Glu Val Val Gly Tyr Ser Glu Glu Arg Leu Lys
145 150 155 160
Leu Tyr Trp Asn Lys Phe Ile Ala Lys Arg Val Val Ser His Asp Arg
165 170 175
Lys Leu Gly Lys Ile Val Arg Glu Lys Tyr Leu Glu Pro Lys Gly Leu
180 185 190
Val Cys Ala Gln Pro Glu Asn Ser Thr Tyr Cys Arg Val Leu Thr Glu
195 200 205
Ile Ile Lys Arg Gln Leu His Ser Gln Ile Glu Lys Ser Lys Phe His
210 215 220
Glu Glu Glu Leu Lys Ser Ile Glu Lys Thr Val Ser Glu Phe Asp Ser
225 230 235 240
Pro Leu Leu Asp Phe Ile Cys Gln Tyr Ala Glu Glu Leu Asn Gln Ile
245 250 255
Asn Ser Gly Leu Ser Lys Tyr Val Ile Lys Asn Ala Val Lys Glu Val
260 265 270
Ile Ser Pro Pro Glu Lys Gln Ser Glu Ile Tyr Val Gln Ser Gln Val
275 280 285
Leu Ser Gln Glu Lys Tyr Lys Pro Leu Val Asn Ala Thr Ile Lys Glu
290 295 300
Ile Leu Ser Gly Tyr Glu Gln Trp Lys Val Lys Ser Arg Tyr Glu Asn
305 310 315 320
Arg Leu Lys Asn Arg Lys Tyr Val Leu Tyr Pro Lys Leu Ser Ala Asn
325 330 335
Tyr Lys Ile Pro Ile Gly Gln Asn Ser Leu Gly Lys Phe Lys Ile Asn
340 345 350
Val Ser Glu Asn Gly Glu Ile Val Ile Arg Leu Asn Asp Met Ala Asp
355 360 365
Val Val Cys Met Pro Ser Lys Tyr Phe Phe Asn Leu Lys Ser Ser Pro
370 375 380
Val Val Asp Lys Lys Lys Gln Leu Val Gly Tyr Gln Ile Ser Phe Asn
385 390 395 400
His Asn Ser Arg Arg Lys Glu Pro Thr Glu Lys Pro Asp Phe Asn Gly
405 410 415
Ile Val Lys Glu Ile Gly Leu Gln Leu Lys Asp Asp Gly Arg Phe Tyr
420 425 430
Ile Thr Leu Pro Tyr Cys Met Glu Tyr Ser Asn Asp Asn Phe Asp Leu
435 440 445
Ile Arg Pro Leu Leu Thr Ser Ser Pro Thr Glu Asp Gln Ile Lys Lys
450 455 460
Met Pro Ser Glu Phe Asn Val Val Gly Phe Asp Leu Asn Leu Ser Met
465 470 475 480
Pro Leu Pro Ile Thr Arg Ala Ile Val Gly Lys Ser Val Lys Gly Glu
485 490 495
Ile Asn Val Glu Tyr Leu Gly Gln Ala Lys Val Ile Glu Ser Thr His
500 505 510
Leu Ile Tyr Asp Asn Asn Arg Cys Lys Val Leu Ile Ala Tyr Lys Arg
515 520 525
Gln Cys Asp Leu Ile Lys Arg Ala Ile Arg Glu Trp Lys Ile Cys Lys
530 535 540
Gly Lys Asn Ile Asp Ile Ser Glu Lys Thr Tyr Glu Trp Leu Glu Ser
545 550 555 560
His Thr Lys Arg Trp Asn Pro Ser Arg Gln Pro Glu Ser Met Gln Asp
565 570 575
Arg Phe Ser Val Ser Lys Met Arg Ile Gln Ile Leu Val Asn Lys Ala
580 585 590
Lys Ser Arg Ile Ala Lys Tyr Asn Asp Asn Ser Trp Lys Thr Gly His
595 600 605
Gly Asn Glu Ser Glu Leu Ile Arg Leu Ile Asp Ala Asp Asp Ala Tyr
610 615 620
Asn Ser Leu Val Ser Thr Tyr Asn Arg Ile His Leu Lys Ser Asn Gln
625 630 635 640
Phe Ile Tyr Ala Leu Pro Ser Lys Asn Asn Ser Arg Ser Asn Lys Lys
645 650 655
Glu Tyr Cys Leu Arg Arg Ile Ala Ala Lys Ile Ala Arg Tyr Cys His
660 665 670
Leu His Asn Val Asn Ile Cys Ile Gly Glu Asn Leu Ser Phe Gln Gln
675 680 685
Asp Ser Asp Asn Ile Ser Lys Asp Asn Ser Leu Val Arg Leu Phe Ser
690 695 700
Ser Lys Ser Ile Ala Asn Tyr Met Lys Leu Ala Met Glu Lys Phe Gly
705 710 715 720
Ile Ala Phe Ile Asp Ser Ala Asp Pro Ser Gly Thr Ser Lys Thr Asp
725 730 735
Pro Val Thr Gly Asn Ile Gly Tyr Arg Asn Lys Phe Asp Lys Arg Lys
740 745 750
Leu His Val Ile Arg Asn Gly Asn Trp Gly Trp Val Asp Ser Asp Ile
755 760 765
Ala Ala Ser Leu Asn Ile Leu Ile Arg Gly Ile Asn Arg Ser Ile Val
770 775 780
Pro Tyr Lys Phe Phe Val Gly Lys Lys Lys Gln Glu Ser Lys Arg Leu
785 790 795 800
Asn His Phe Leu Asn Lys Ile Phe Gly Thr Thr Lys Val Phe Phe Tyr
805 810 815
Glu Asp Gln Phe Gly Phe Ala Asn Pro Ser Leu Ser Lys Lys Glu Gly
820 825 830
Glu Asn Leu Ile Ala Asn Gln Tyr Leu Tyr Tyr Arg Glu Gly Lys Phe
835 840 845
Val Thr Gln Lys Ile His Arg Gln Ile Glu Asp Asp Phe Lys Lys Ile
850 855 860
Asp Phe Ser Asn Thr Pro Glu Val Asn Leu Ile Pro Ser Gly Val Lys
865 870 875 880
Leu Lys Asn Phe Gln Phe Glu Ser Arg Ala Asp Pro Lys Lys Lys Arg
885 890 895
Lys Val
<210> 77
<211> 932
<212> PRT
<213> 人工序列
<220>
<223> Cas12h.2-NLS融合蛋白的氨基酸序列
<400> 77
Met Ala Thr Arg Ser Phe Ile Arg Thr Gly Asn Leu Lys Ala Lys Asn
1 5 10 15
Thr Ala Glu Glu Val Met Gln Trp Tyr Ala Asp Leu Gln Ser Asp Tyr
20 25 30
Arg Ser Phe Leu Asn Leu Phe Phe Gly Trp Met Ala Ile Gly Tyr Gly
35 40 45
Thr Asn Ala Glu Asp Glu Val Phe Tyr Thr Ser Lys Glu Glu Ser Glu
50 55 60
Arg Leu Arg Ser Leu Thr Ile Gly Asp Ala Lys Lys Glu Gln Leu Ala
65 70 75 80
Val Ser Phe Ile Glu Leu Leu Leu Lys Gly Gly Glu Asn Ala Ser Ser
85 90 95
Cys Tyr Asn Val Phe Tyr Arg Asn Tyr Lys Ser Leu Gly Lys Ala Lys
100 105 110
Leu Thr Gln Lys Lys Asn Asp Phe Leu Ser Ala Leu Pro Leu Leu Asp
115 120 125
Glu Asn Lys Ile Lys Glu Tyr Phe Lys Thr Asp Glu Gln Leu Ser Gln
130 135 140
Ile Cys Ile Glu Glu Trp Leu Glu Tyr Gly Val Lys Asn Leu Pro Leu
145 150 155 160
Pro Glu Ile Trp Ala Glu Val Ser Pro Arg Leu Ala Ser Ile Glu Arg
165 170 175
Ser Leu Gly Val Asp Leu Arg Leu Ala Phe Gly Leu Ser Cys Ile Arg
180 185 190
Ser Arg Asp Cys Asn Tyr Cys Arg Ile Leu Ile Glu Met Val Gly Arg
195 200 205
Asp Leu Arg Ser Ile Phe Glu Lys Tyr Asn Asn His Leu Leu Glu Thr
210 215 220
Glu Lys Ile Lys Leu Ser Met Asn Asp Lys Gln Gly Pro Val Tyr Asp
225 230 235 240
Ser Ile Cys Cys Phe Ala Ala Glu Leu Glu Ser Lys Asn Ser Gly Leu
245 250 255
Thr Lys Tyr Val Leu Thr Lys Gly Ile Asp His Val Lys Lys Gly Thr
260 265 270
Gly Glu Lys Thr Asp Ile Arg Leu Ala Val Lys Glu Leu Lys Lys Asn
275 280 285
Lys Tyr Arg Ile Leu Ile Glu Ser Ser Tyr Ser Glu Ile Met Ser Ala
290 295 300
Tyr Ser Cys Trp Arg Thr Lys Lys Gln Leu Glu Lys Arg Lys Leu Tyr
305 310 315 320
Pro Cys Phe Asp Pro Asn Arg Asn Asp Tyr Lys Val Pro Val Gly Gln
325 330 335
Gly Ser Leu Gly Asn Phe Thr Val Ser Val Glu Asp Ser Gly Asp Val
340 345 350
Leu Ile Glu Ile Val Gly Val Gly Val Ile Arg Cys Ala Ala Ser Cys
355 360 365
Tyr Phe Ser Gly Ile Val Phe Asp Glu Ile Arg Asn Lys Asn Gly Arg
370 375 380
Thr Gly Tyr Ser Leu Asn Phe Cys His Lys Ser Ile Ser Lys Gly Lys
385 390 395 400
Lys Ala Val Lys Ala Ala Ser His Thr Gly Asp Lys Ile Ser Gly Val
405 410 415
Leu Lys Glu Ile Gly Leu Arg Asn Thr Asp Ser Gly Phe Phe Val Ser
420 425 430
Leu Pro Tyr Ser Ile His His Asp Glu Lys Asn Phe Lys Ile Ala Glu
435 440 445
Phe Phe Met Ser Ala Cys Pro Lys Lys Glu Asn Val Glu Asn Leu Pro
450 455 460
Asp Lys Ile Val Val Gly Ala Ile Asp Leu Asn Val Ser Asn Pro Val
465 470 475 480
Ala Ala Val Lys Ala Val Val Tyr Arg Asp Asp Lys Ser Gly Gln Leu
485 490 495
Asn Ala Leu Asp Tyr Gly Ser Gly Asn Leu Ile Lys Lys Pro Phe Met
500 505 510
Leu Val Ala Asn Gly Pro Arg Ile Lys Asn Leu Ile Glu Ile Arg Asp
515 520 525
Asp Ala Arg Arg Val Ile Gly Ala Ile Arg Glu Phe Lys Val Ser Asn
530 535 540
Ala Val Lys Glu His Val Gly Glu Asp Thr Arg Asp Phe Leu Ile Leu
545 550 555 560
Cys Gly Asp Thr Lys Ser Ser Ser Thr Arg Tyr Leu Ile Gln Ser Trp
565 570 575
Val Lys Lys Ile Asn Ser Arg Leu Arg Lys Ile Lys Phe Glu Met Arg
580 585 590
Ser Gly Gly Tyr Arg Asp Cys Ala Asp Asn Ile Arg Leu Ile Glu Ala
595 600 605
Met Asp Gln Cys Ala Ser Met Ala Glu Ser Tyr Asn Arg Ile His Leu
610 615 620
Lys Ser Gly Glu Lys Leu Val Lys Val Ala Lys Phe Asp Lys Ser Arg
625 630 635 640
Ala Asn Phe Arg Asn Phe Val Leu Arg Gln Leu Ala Ser Lys Ile Ala
645 650 655
Asn Glu Met Lys Asp Cys Asn Val Val Phe Gly Glu Asp Leu Asp Phe
660 665 670
Ile Phe Asp Ser Asp Lys Asn Asn Asn Ala Leu Leu Arg Leu Phe Ser
675 680 685
Ala Ala Thr Leu Leu Lys Tyr Ile Ile Glu Ala Leu Glu Lys Ile Gly
690 695 700
Val Gly Phe Val Lys Val Ala Lys Asn Gly Thr Ser Gln Ser Asp Pro
705 710 715 720
Val Thr Ser Asn Pro Gly Trp Arg Asp Asp Lys Asn Lys Ser Arg Leu
725 730 735
Tyr Val Val Arg Asp Lys Gln Leu Gly Trp Ile Asp Ser Asp Leu Ala
740 745 750
Ala Thr Met Asn Ile Leu Ile Gln Gly Leu Asn His Ser Val Cys Pro
755 760 765
Tyr Lys Phe Tyr Val Lys Glu Tyr Glu Asn Lys Pro Asn Ser Thr Gln
770 775 780
Asp Ser Ile Asn Ala Ile Lys Lys Pro Glu Glu Ala Ile Gly Lys Arg
785 790 795 800
Ile Lys Arg Phe Phe Asn Leu Lys Tyr Gly Ser Ser Val Pro Lys Phe
805 810 815
Val Ser Asp Asp Arg Gly Arg Val Thr Phe Ala Lys Lys Ile Asp Ser
820 825 830
Thr Gln Thr Arg Leu Ile Asn Gln Phe Val Tyr Ala His Ser Ser Cys
835 840 845
Ile Val Thr Cys Glu Leu His Asn Glu Met Val Asn Lys Ile Lys Gln
850 855 860
Leu Ala Val Glu Lys Pro Asn Cys Gln Glu Phe Asp Val Thr Cys Asp
865 870 875 880
Pro Asp Gly Arg Tyr Asn Asn Phe Ala Leu Pro Glu Val His Asp Ser
885 890 895
Ser Lys Asp Val Gly Ala Lys Ala Leu Thr Thr Lys Asp Val Asp Phe
900 905 910
Lys Thr Ile Leu Lys Asp His Thr Ala Ser Arg Ala Asp Pro Lys Lys
915 920 925
Lys Arg Lys Val
930
<210> 78
<211> 1308
<212> PRT
<213> 人工序列
<220>
<223> Cas12w.1-NLS融合蛋白的氨基酸序列
<400> 78
Met Gly Lys Asn Glu Asn Lys Tyr Gln Leu Ser Lys Thr Leu Arg Phe
1 5 10 15
Gly Leu Thr Leu Lys Glu Lys Ile Ser Asn Asn Glu Lys Thr Pro Tyr
20 25 30
Gln Ser His Ser Gln Phe Arg Asp Leu Ile Ile Leu Ser Glu Asn Arg
35 40 45
Ile Arg Glu Gly Ile Ser Thr Pro Gln Asn Arg Asp Leu Pro Ser Phe
50 55 60
Ile His Arg Ile Gln Asn Cys Thr Asp Phe Ile Asn Asp Phe Ile His
65 70 75 80
Asp Trp Trp Met Ile Leu Met His Thr Gly Gln Ile Glu Leu Asp Lys
85 90 95
Asp Tyr Tyr Lys Ser Leu Thr Lys Lys Val Gly Phe Val Gly Phe Trp
100 105 110
Tyr Lys Glu Asn Lys Lys Lys Gly Gly Lys Thr Lys Gln Pro Gln Ala
115 120 125
Arg Asn Ile Pro Met Gly Glu Leu Arg His Leu Cys Pro Gln Asn Thr
130 135 140
Lys Glu Cys Ala Thr Tyr Ile Thr Asp Tyr Trp Lys Asp Leu Leu Ile
145 150 155 160
Thr Ala Thr Asn Lys Leu Tyr Glu Ser Ser Glu Gln Gln Lys Lys Phe
165 170 175
Ile Lys Ala Met Glu Gln Asn Arg Thr Asp Asn Lys Pro Asn Glu Ile
180 185 190
Asp Leu Lys Lys Ser Phe Leu Ser Leu Val Ser Val Thr Met Glu Leu
195 200 205
Leu Asn Pro Ile Leu Asn Gly Gln Ile Leu Phe Asn Lys Met Asp Arg
210 215 220
Leu Asp Met Ser Lys Lys Ser Asp Asn Asp Phe Ile Asp Phe Val Asn
225 230 235 240
Asp His Glu Thr Val Arg Glu Leu Asn Asn Asp Ile Glu Glu Ile Ile
245 250 255
Ala Asp Phe Lys Glu Asn Gly Asn Asn Val Asn Tyr Cys Lys Ala Thr
260 265 270
Leu Asn Pro Asp Thr Ala Leu Lys Gln His Asn Asn Asn Ile Pro Asn
275 280 285
Asp Ile Ala Thr Asp Leu Glu Glu Leu Met Met Asp Ser Ile Val Gly
290 295 300
Asn Tyr Asp Asp Val Asn Ser Phe Met Asp Asn Tyr Val Ser Asn Leu
305 310 315 320
Ser Ala Lys Asp Lys Ile Lys Lys Ile Lys Asp Ser Asn Ile Ser Leu
325 330 335
Ile Tyr Arg Ala Ile Leu Phe Lys Tyr Lys Met Ile Pro Ala Asn Val
340 345 350
Arg Arg Asp Ile Ala Gln Gly Met Ala Lys Lys Leu Asn Lys Asp Glu
355 360 365
Glu Asn Ile Tyr Ser Phe Leu Cys Glu Phe Gly Thr Leu Arg Thr Pro
370 375 380
Gln Lys Asp Tyr Ala Asp Leu Lys Asp Lys Asp Ser Phe Asn Leu Asp
385 390 395 400
Asn Tyr Pro Leu Lys Val Ala Phe Asp Phe Ala Trp Glu Gly Leu Ala
405 410 415
Lys Ala Trp Tyr His Asp Gln Ser Asp Phe Pro Ile Asp Pro Cys Arg
420 425 430
Asp Phe Leu Gln Glu Asn Phe Asp Val Asn Leu Glu Glu Asp Gln Glu
435 440 445
Asp Glu Tyr Phe Leu Leu Tyr Ala Asp Leu Ile Glu Leu Asn Ala Leu
450 455 460
Leu Ser Thr Leu Asp Lys Gly Asn Pro Ala Asp Pro Asp Ser Ile Lys
465 470 475 480
Asn Glu Ala Leu Glu Met Val Glu Tyr Ile Asn Trp Asn Ser Leu Asp
485 490 495
Lys Lys Asn Gly Asn Tyr Tyr Lys Lys Ile Ile Lys Asn Arg Leu Lys
500 505 510
Ser Ser Lys Gly Asn Glu Thr Tyr Glu Arg Ile Lys Lys Glu Ile Ser
515 520 525
Met Ser Arg Gly Arg Leu Lys Asn Lys Ile Glu Lys Tyr Asp Asp Leu
530 535 540
Thr Ser Gln Tyr Lys Arg Ile Ala Met Asp Leu Gly Lys Lys Phe Ala
545 550 555 560
Ser Leu Arg Asp Lys Ile Ile Ala Ala Asn Glu Asp Asn Lys Val Thr
565 570 575
His Tyr Ala Met Ile Leu Glu Asp Ser Asn Cys Asp Lys Tyr Leu Leu
580 585 590
Leu Gln Lys Val Ser Asn Asn Ile Tyr His Cys Met Ser Tyr Asp Ser
595 600 605
Ser Asp Pro Lys Ala Tyr Tyr Val Asp Ser Ile Thr Ser Ser Ala Ile
610 615 620
Ala Lys Met Ile Arg Lys Glu Thr Asn Pro Ser Lys Ile Arg Glu Tyr
625 630 635 640
Ala Glu Leu Glu Glu Lys Glu Arg Glu Arg Arg Asn Val Asp Asp Trp
645 650 655
Cys Arg Phe Ile Ser Lys Lys Glu Tyr Asp Arg Arg Tyr Gln Leu Asn
660 665 670
Ile Asn Asn Gly Leu Ser Phe Glu Ala Leu Lys Lys Glu Ile Asp Ser
675 680 685
Lys Ser Tyr Ile Leu Val Lys Lys Asn Ile Ser Val Asp Ser Ile Arg
690 695 700
Glu Leu Val Glu Asn Glu Gly Cys Leu Leu Phe Pro Ile Val Asn Lys
705 710 715 720
Asp Leu Thr Lys Glu Arg Lys Thr Thr Glu Asp Asn Gln Phe Thr Lys
725 730 735
Asp Trp Asn Met Ile Phe Ser Gly Ser Glu Thr Asn Trp Arg Leu Thr
740 745 750
Pro Glu Phe Arg Val Thr Tyr Arg Asn Pro Val Pro Gly Tyr Pro Asn
755 760 765
Asp Lys Phe Gly Ser Lys Arg Tyr Ser Arg Phe Gln Met Asn Ala His
770 775 780
Phe Val Cys Asp Phe Ile Pro Ser Ser Asn Ser Tyr Thr Ser Asn Arg
785 790 795 800
Glu Gln Ile Ala Ile Phe Lys Asp Glu Gly Glu Gln Lys Lys Arg Val
805 810 815
Glu Glu Phe Asn Arg Thr Leu Ser Asn Ile Asn Gln Lys Phe Tyr Val
820 825 830
Ile Gly Ile Asp Arg Gly Gln Lys Glu Leu Ala Thr Leu Cys Val Val
835 840 845
Asp Gln Asp Lys Lys Ile His Gly Asp Phe Lys Ile Tyr Thr Arg Lys
850 855 860
Phe Asn Ser Glu Arg Lys Gln Trp Glu His Tyr Ser Leu Glu Gly Glu
865 870 875 880
Lys Gly Thr Arg Asn Ile Leu Asp Leu Ser Asn Leu Arg Val Glu Thr
885 890 895
Thr Ile Ile Ile Asp Gly Lys Pro Glu Arg Arg Gln Val Leu Val Asp
900 905 910
Leu Ser Glu Val Leu Val Lys Asp Lys Glu Gly Asn Tyr Thr Lys Pro
915 920 925
Asn Lys Met Gln Ile Lys Met Gln Gln Met Ala Tyr Val Arg Lys Leu
930 935 940
Gln Phe Gln Met Gln Ala Asn Pro Thr Glu Val Leu Glu Trp Tyr Glu
945 950 955 960
Gln Asn Pro Thr Glu Glu Leu Ile Ile Lys Asn Leu Val Asp Lys Glu
965 970 975
Asn Gly Glu Lys Gly Leu Ile Ser Phe Tyr Gly Thr Ala Leu Val Glu
980 985 990
Leu Asp Gln Thr Leu Pro Val Ser Lys Ile Lys Glu Met Leu Glu Glu
995 1000 1005
Phe Lys Ile Leu Lys Gln Arg Glu Ser Lys Lys Glu Asn Val Gln
1010 1015 1020
Lys Glu Leu Asn Asn Leu Thr Gln Leu Glu Ala Val Asp Ser Leu
1025 1030 1035
Lys Ala Gly Ile Val Ala Asn Met Val Gly Val Ile Ser Tyr Ile
1040 1045 1050
Leu Lys Thr Leu Asp Tyr Asn Ala Tyr Ile Ser Leu Glu Asp Leu
1055 1060 1065
Ser Thr Val Gln Ser Ser Thr Glu Phe Ala Ser Gly Ile Ser Gly
1070 1075 1080
Ala Ile Thr Lys Met Ser Arg Glu Glu Gly Arg Arg Ile Asp Val
1085 1090 1095
Glu Lys Tyr Ala Gly Leu Gly Leu Tyr Asn Phe Phe Glu Met Gln
1100 1105 1110
Leu Leu Arg Lys Leu His Arg Ile Gln Thr Asp Asn Gly Asn Ile
1115 1120 1125
Leu His Leu Val Pro Ala Phe Arg Ala Gln Lys Asn Tyr Asp His
1130 1135 1140
Ile Met Val Gly Lys Glu Lys Ile Lys Asn Gln Phe Gly Ile Val
1145 1150 1155
Phe Phe Val Asp Ala Ala Ala Thr Ser Ile Lys Cys Pro Arg Cys
1160 1165 1170
Gly Ala Val Asn Glu Asp Lys Phe Asn Pro Asp Lys Gln Lys Tyr
1175 1180 1185
Pro Asp Ala Glu Lys Gly Pro Lys Leu Arg Asn Arg Lys Glu Gln
1190 1195 1200
Ser Gly Lys Lys Val Trp Val Thr Arg Asp Lys Glu Asp Asp Asp
1205 1210 1215
Arg Ile Lys Cys Tyr Cys Cys Gly Phe Asp Thr Lys Glu Lys Asn
1220 1225 1230
Glu Gly Asn Pro Phe Met Tyr Ile Lys Ser Gly Asp Asp Asn Ala
1235 1240 1245
Ala Tyr Leu Ile Ser Asp Leu Gly Val Glu Ser Tyr Arg Lys Ala
1250 1255 1260
Tyr Glu Leu Ala Ala Thr Val Val Glu Asp Arg Lys Lys Thr Leu
1265 1270 1275
Thr Asn Asn Leu Asn Gln Ser Asn Tyr Lys Ile Arg Phe Leu Trp
1280 1285 1290
His Thr Met Tyr Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val
1295 1300 1305
<210> 79
<211> 1268
<212> PRT
<213> 人工序列
<220>
<223> Cas12w.2-NLS融合蛋白的氨基酸序列
<400> 79
Met Asp Ala Asp Lys Thr Thr Lys Ala Ile Asn Glu Tyr Gln Thr Gln
1 5 10 15
Lys Thr Ile Arg Phe Gly Leu Thr Ala Thr Asn Gln Asn Leu Tyr Ser
20 25 30
Glu Glu Ile Met Lys Leu Leu Asn Ile Ser Glu Glu Arg Ile Ile Lys
35 40 45
Glu Lys Val Lys Val Asn Asn Asp Thr Asp Lys Thr Asn Gln Leu Arg
50 55 60
Gly Cys Leu Val Gln Ile Lys Lys Tyr Leu Lys Thr Trp Glu Asn Ile
65 70 75 80
Tyr Ala Gln Ile Asp Phe Leu Ala Ile Thr Lys Asp Tyr Tyr Lys Val
85 90 95
Ile Ser Lys Lys Ala Arg Phe Asp Phe Asp Lys Gly Asn Gly Ser Glu
100 105 110
Ile Lys Leu Ser Ser Leu Gln Ser Thr His Asn Lys Lys Lys Arg Tyr
115 120 125
Gln Tyr Ile Ile Asp Phe Trp Lys Glu Asn Leu Arg Lys Thr Glu Asn
130 135 140
Leu Tyr Arg Lys Ser Asp Asp Leu Leu Lys Ile Phe Glu Glu Ala Lys
145 150 155 160
Asn Gln Asn Arg Asp Asp Lys Lys Leu Asn Lys Val Glu Leu Arg Lys
165 170 175
Thr Phe Leu Asn Leu Phe Thr Leu Val Asn Glu Ser Leu Lys Pro Leu
180 185 190
Ile Glu Gly Asn Leu Phe Ile Val Asn Asp Asp Lys Ile Asp Glu Lys
195 200 205
Asn Ser Lys His Asn Tyr Val Phe Tyr Phe Ile Ser Lys Thr Glu Glu
210 215 220
Arg Arg Leu Leu Tyr Asp Asn Ile Cys Thr Leu Gln Asp Tyr Phe Lys
225 230 235 240
Asn Asn Gly Gly Tyr Val Pro Phe Gly Arg Val Thr Leu Asn Lys Trp
245 250 255
Thr Ala Leu Gln Lys Phe Asn Asn Arg Asp Ile Glu Ile Asn Arg Ile
260 265 270
Ile Lys Glu Leu Lys Ile Asn Asn Ile Ser Thr Gln Lys Thr Asp Tyr
275 280 285
Lys Tyr Asn Asp Phe Thr Glu Asn Phe Lys Glu Lys Lys Asp Glu Asn
290 295 300
Gly Lys Val Val Lys Asn Ser Ala Gly Asn Ile Ile Trp Asp Leu Lys
305 310 315 320
Ala Asn Ala Lys Ser Val Ile Glu Ile Cys Gln Phe Phe Lys Tyr Lys
325 330 335
Lys Val Pro Ile Asn Ala Arg Leu Asn Leu Ala Lys Arg Leu Ile Lys
340 345 350
Asp Asn Lys Leu Lys Lys Glu Gln Glu Asn Thr Phe Leu Ser Glu Phe
355 360 365
Gly Val Leu Lys Thr Pro Ala Phe Asp Tyr Ala Arg Asp Lys Glu Asn
370 375 380
Phe Asn Leu Thr Asn Tyr Pro Leu Lys Val Ala Phe Asp Tyr Ala Trp
385 390 395 400
Glu Asn Cys Ala Lys Asp Lys Tyr Glu Lys Ile Pro Phe Pro Lys Glu
405 410 415
Gln Cys Glu Arg Tyr Leu Gln Thr Ala Phe Glu Ile Asp Ala Thr Lys
420 425 430
Asp Glu Asn Lys Lys Leu Ile Asp Thr His Leu Asn Lys Tyr Ala Asp
435 440 445
Leu Leu Gln Phe Lys Ile Leu Leu Glu Arg Phe Lys Ala Glu Phe His
450 455 460
Lys Thr Asn Glu Glu Thr Asn Lys Asn Asn Ile Gln Lys Leu Arg Asn
465 470 475 480
Val Phe Ser Gly Leu Asp Tyr His Gly Asp Asn Arg Leu Asn Lys Asn
485 490 495
Gln Ile Gln Lys Ala Ile Glu Ala Trp Phe Asp Asn Lys Glu Gln Asn
500 505 510
Ile Gly Lys Lys Lys Glu Asn Glu Lys Leu Leu Thr Glu Asn Glu Lys
515 520 525
Asn Asn Phe Ser Leu Ser Met Gln Ile Ile Gly Gln Glu Arg Gly Gly
530 535 540
Leu Lys Asn Gly Ile Pro Lys Tyr Lys Glu Leu Thr Glu Met Phe Lys
545 550 555 560
Val Cys Ala Ser Lys Phe Gly Lys Gln Phe Ala Asp Leu Arg Asp Tyr
565 570 575
Phe Asn Glu Ala Tyr Glu Val Asp Lys Ile Lys Tyr Arg Ala Trp Ile
580 585 590
Ile Glu Asp Asp Lys Lys Asn Arg Phe Val Leu Phe Val Asn Lys Glu
595 600 605
Lys Ala Phe Asp Leu Thr Ser Glu Glu Gly Asp Leu Trp Phe Tyr Glu
610 615 620
Val Lys Ser Leu Thr Ser Lys Ser Leu Val Lys Phe Ile Lys Asn Arg
625 630 635 640
Gly Ala Tyr Pro Asp Phe His Asp Val Lys Asn Ser Phe His Tyr Ser
645 650 655
Ser Ile Lys Lys Asp Trp Gln Asn Tyr Lys Asn Asp Pro Glu Phe Leu
660 665 670
Asp Lys Leu Lys Glu Cys Leu Lys Asn Ser Lys Ile Ala Lys Asp Gln
675 680 685
Lys Trp Ala Lys Phe Cys Trp Asp Phe Lys Gln Cys Asp Thr Tyr Glu
690 695 700
Lys Leu Glu Lys Glu Val Asp Arg Lys Gly Tyr Lys Leu Glu Gly Cys
705 710 715 720
Lys Ser Glu Pro Lys Thr Ile Ser Leu Thr Gln Leu Thr Asp Trp Val
725 730 735
Glu Asn Lys Asp Cys Phe Leu Leu Pro Ile Val Asn Gln Asp Ile Asn
740 745 750
Lys Gly Asp Lys Arg Thr Lys Asn Gln Asn Gln Phe Thr Lys Asp Trp
755 760 765
Phe Asp Ile Phe Glu Asn Lys Lys Arg Leu His Pro Glu Phe Asn Ile
770 775 780
Phe Tyr Arg Phe Pro Thr Lys Asp Tyr Pro Asn Thr Lys Phe Lys Asn
785 790 795 800
Gly Thr Glu Lys Thr Lys Arg Tyr Ser Arg Phe Gln Met Leu Ala Tyr
805 810 815
Phe Gly Cys Glu Val Ile Pro Ser Gly Asn His Leu Ser Lys Lys Glu
820 825 830
Gln Ile Ala Ile Phe Asn Asn Asp Lys Lys Gln Lys Glu Glu Val Glu
835 840 845
Lys Tyr Asn Lys Ser Ile Ser Ser Asp Cys Asp Tyr Val Ile Gly Ile
850 855 860
Asp Arg Gly Ile Lys Gln Leu Ala Thr Leu Cys Val Leu Asp Lys Asn
865 870 875 880
Gly Val Ile Gln Gly Asp Phe Gln Ile Phe Thr Arg Thr Phe Asn Lys
885 890 895
Gln Thr Lys Gln Trp Glu His Lys Glu Leu Glu Gln Arg Asn Ile Leu
900 905 910
Asp Leu Ser Asn Leu Arg Val Glu Thr Thr Ile Thr Gly Lys Lys Val
915 920 925
Leu Val Asp Leu Ser Lys Ile Lys Asp Asp Glu Gly Asn Tyr Thr Asn
930 935 940
Leu Lys Gln Thr Ile Lys Leu Lys Gln Leu Ala Tyr Ile Arg Glu Leu
945 950 955 960
Gln Tyr Ala Met Gln Thr Arg Pro Asp Asp Leu Leu Asp Phe Val Lys
965 970 975
Ser Ile Asn Ser Ala Asn Asp Ile Thr Ala Glu Asn Ile Lys His Phe
980 985 990
Ile Ser Pro Tyr Lys Glu Gly Lys Asn Tyr Asp Asp Leu Pro Lys Val
995 1000 1005
Glu Met Phe Asn Leu Leu Lys Glu Trp Gly Asn Ala Asp Glu Asn
1010 1015 1020
Gly Lys Arg Lys Ile Ala Glu Leu Asp Pro Ala Asp Asn Leu Lys
1025 1030 1035
Ser Gly Ile Val Ala Asn Met Val Gly Val Val Ala Phe Leu Cys
1040 1045 1050
Glu Asn Tyr Asn Tyr Lys Val Arg Ile Ala Leu Glu Asp Leu Thr
1055 1060 1065
Arg Ala Tyr Gly Ile Gln Lys Asp Ala Leu Asn Gly Thr Ala Ile
1070 1075 1080
Tyr Gln Asn Asp Glu Asp Phe Lys Glu Gln Glu Asn Arg Arg Leu
1085 1090 1095
Ala Gly Val Gly Thr Met Gln Phe Phe Glu Val Gln Leu Leu Arg
1100 1105 1110
Lys Leu Phe Lys Ile Gln Val Asp Lys Asn Leu His Leu Ile Pro
1115 1120 1125
Ala Phe Arg Ser Val Asp Asn Tyr Glu Lys Ile Val Arg Arg Asp
1130 1135 1140
Lys Gln Asn Ser Gly Asp Glu Phe Val Asn Tyr Pro Phe Gly Ile
1145 1150 1155
Val Cys Phe Val Asp Pro Lys Tyr Thr Ser Gln Gln Cys Pro Tyr
1160 1165 1170
Cys Asn Asn Thr His Lys His Lys Lys Asn Asp Thr Glu Thr Gly
1175 1180 1185
Lys Lys Ala Phe Tyr Arg Asn Lys Gly Glu Asn Lys Asn Ser Leu
1190 1195 1200
Leu Cys Glu Lys Cys Gly Val Ser Thr Ile Glu Gly Glu Glu Thr
1205 1210 1215
Leu Ser Ser Lys Asn Asp Asn Lys Lys Gln Phe Asn Ile His Tyr
1220 1225 1230
Ile Thr Asp Gly Asp Gln Asn Gly Ala Tyr His Ile Ala Asn Lys
1235 1240 1245
Val Val Ile Asn Phe Gln Lys Asp Ser Ser Arg Ala Asp Pro Lys
1250 1255 1260
Lys Lys Arg Lys Val
1265
<210> 80
<211> 1022
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.1-NLS融合蛋白的氨基酸序列
<400> 80
Met Asp Tyr Gln Gln Tyr Glu Phe Thr Arg Thr Ile Arg Phe Asn Leu
1 5 10 15
Ser Gly Asp Asp Lys Arg Ala Leu Met Leu Asp Leu Leu Asp Asp Thr
20 25 30
Gln Glu Gly Met Leu Ala Ala Phe Gln Glu Thr Tyr Lys Asn Leu Leu
35 40 45
Phe Ala Phe Gln Glu Ala Ile Leu Arg Ala Asp Gly Ser Gly Asn Leu
50 55 60
Arg Val Gly Arg Leu Glu Ile Lys Lys Ser Trp Leu Arg Gln Tyr Ala
65 70 75 80
Arg Glu Tyr Phe Tyr Ala Leu Ser Glu Asp Glu Arg Arg Cys Lys Asn
85 90 95
Lys Phe Gln Ala Lys Leu Phe Asp Arg Val Leu Ser Asp Trp Leu Glu
100 105 110
Arg Asn Asn Glu Leu Leu Gln Arg Leu Asn Asn Ile Leu Ser Leu Pro
115 120 125
Gln Glu Ser Lys Thr Gly Ala Ser Asp Leu Ser Leu Leu Val Arg Gln
130 135 140
Leu Lys Gly Ala Glu Tyr Phe Tyr Phe Ile Arg Asp Phe Thr Gln Ser
145 150 155 160
Gly Ile Ile Asn Asp Lys Asp Ser Asp Glu His Ile Lys Asn Leu Ala
165 170 175
Gly Ile Val Glu Lys Phe Glu Thr Leu Leu Asp Lys Val Leu Phe Leu
180 185 190
Thr Ala Pro Asn Ser Ser Gln Gly Val Glu Thr Thr Arg Ala Ser Phe
195 200 205
Asn Tyr Tyr Thr Val Asn Lys Ile Ser Lys Asn Phe Asp Glu Asn Ile
210 215 220
Lys Lys Ala Asn Gly Arg Leu Cys Ser Ser Tyr Gln Asn Ser Met Asn
225 230 235 240
Glu Glu Leu Leu Arg Lys Val Gly Phe Leu Lys Tyr Leu Lys Asp Glu
245 250 255
Tyr Arg Ala Glu Leu Gln Asn Val Ser Leu Lys Asp Leu Tyr Glu Ala
260 265 270
Leu Lys Lys Phe Lys Ser Gln Gln Lys Thr Ala Phe Ile Gln Ala Val
275 280 285
Gln Lys Asn Lys Ser Glu Lys Glu Leu Met Arg Glu Phe Pro Leu Phe
290 295 300
Asn Gly Lys Gln Pro Asp Thr Leu Gln Lys Phe Ile Leu Glu Thr Asp
305 310 315 320
Lys Ile Lys Arg Gly Ala Tyr Phe Gln Lys Trp Gly Phe Asp Asn Tyr
325 330 335
Ile Ser Phe Cys Asn Lys Ile Phe Lys Pro Val Ala Met Glu Thr Gly
340 345 350
Thr Arg Lys Ala Lys Ile Arg Ala Leu Glu Gln Glu Lys Ile Glu Ala
355 360 365
Arg Leu Leu Gln Tyr Trp Ala His Ile Leu Val Lys Asp Gly Lys Tyr
370 375 380
Phe Leu Leu Leu Ile Pro Lys Glu Lys Met Gly Glu Ala Lys Val Phe
385 390 395 400
Phe Ala Arg Leu Ser Asp Gln Glu Gly Gly Glu Tyr Thr Leu Tyr Ala
405 410 415
Phe Asn Ser Leu Thr Leu Arg Ala Leu Lys Lys Leu Ile Arg Arg Asn
420 425 430
Leu Gly Lys Glu Gln Val Arg Leu Ser Ala Gly Asp Ala Asp Ala Ile
435 440 445
Ala Leu Cys Gln Glu Val Leu Arg Gly Arg Tyr His Gln Leu Lys Asp
450 455 460
Leu Asp Leu Ser Gly Phe Glu Lys Glu Ile Ala Glu Ile Ala Asn Thr
465 470 475 480
Gln Tyr Glu Asn Glu Glu Glu Phe Arg Ile Ala Leu Glu Gln Val Ala
485 490 495
Tyr Tyr Leu Ser Glu Arg Lys Met Asn Glu Glu Ser Ile Glu Tyr Leu
500 505 510
Lys Lys Asn Leu Gly Ala Ile Leu Leu Glu Ile Ser Ser Tyr Asp Leu
515 520 525
Glu Arg Asn Ile Thr Gly Glu Ser Lys Glu His Thr Arg Leu Trp Ser
530 535 540
Asp Phe Trp Asn Pro Asn Asn Lys Lys Glu Cys Phe Ser Thr Arg Leu
545 550 555 560
Asn Pro Glu Leu Arg Ile Phe Tyr Arg Pro Pro Arg Glu Gln Lys Asp
565 570 575
Pro Lys Lys Gln Lys Asn Arg Phe Ser Lys Asp His Leu Ala Val Ala
580 585 590
Phe Thr Ile Ala Gln Asn Ala Ala Arg Lys Arg Met Glu Thr Ser Phe
595 600 605
Ala Glu Glu Lys Asp Leu Val Glu Gln Val Lys Lys Phe Asn Glu Glu
610 615 620
Val Val Gly Lys Phe Ile Asp Glu Lys Ser Asp Asn Leu Tyr Tyr Tyr
625 630 635 640
Gly Ile Asp Arg Gly Gln Gln Glu Leu Ala Thr Leu Cys Val Val Arg
645 650 655
Phe Ser Lys Glu His Tyr Glu Ala Met Leu Glu Asp Asn Phe Ile Lys
660 665 670
Lys Phe Ser Lys Pro Ile Pro Ala Gln Ile Thr Ala Tyr Arg Ile Lys
675 680 685
Asp Glu His Met Ser Tyr Arg Lys Asn Ile Thr Arg Asp Leu Lys Gly
690 695 700
Asn Glu Thr Glu Glu Ile Leu Phe Lys Asn Pro Ser His Phe Ile Asp
705 710 715 720
Glu Val Glu Asn Phe Glu Glu Val Ser Thr Pro Cys Ile Asp Leu Thr
725 730 735
Thr Ala Lys Leu Ile Lys Gly Lys Ile Ile Leu Asn Gly Asp Ile Gln
740 745 750
Thr Tyr Leu Ala Leu Lys Lys Ala Asn Gly Lys Arg Gln Leu Phe Glu
755 760 765
Lys Phe Ala Lys Ile Asp Asp Ser Ala Lys Ile Glu Phe Asp Asp Ser
770 775 780
Glu Gly Arg Phe Gln Val Lys Ser Lys Ala Thr Glu Arg Glu Glu Tyr
785 790 795 800
Gln Phe Leu Pro Tyr Tyr Gly Pro Glu Gln Glu Asn Ile Ser Pro Arg
805 810 815
Glu Asp Met Arg Arg Glu Leu Gln Ala Tyr Leu Asp Lys Leu Arg Ser
820 825 830
Ser Glu Ser Phe Glu Glu Asp Ile Ser Ile Glu Lys Ile Asn His Leu
835 840 845
Arg Asp Ala Ile Thr Ser Asn Met Val Gly Ile Ile Ala Phe Leu Phe
850 855 860
Thr Glu Tyr Pro Gly Ile Ile Asn Leu Glu Asn Leu His Ser Arg Glu
865 870 875 880
Asn Ile Glu Lys Asn Trp Arg Lys Asn Asn Glu Asp Ile Ser Arg Arg
885 890 895
Leu Glu Trp Gly Leu Tyr Lys Lys Phe Gln Lys Ile Gly Leu Val Pro
900 905 910
Pro Arg Leu Arg Gln Thr Val Leu Leu Arg Glu Asn Glu Thr Glu Arg
915 920 925
Gln Glu Lys Leu Asn Gln Phe Gly Ile Ile His Phe Ile Pro Thr Glu
930 935 940
Lys Thr Ser Ala Arg Cys Pro Tyr Cys Gly Glu Asn Thr Pro Met Lys
945 950 955 960
Gln Arg Asn Glu Asp Lys Phe Lys Leu His Ala Tyr Ile Cys Arg Ser
965 970 975
Asn Glu Glu Asn Cys Gly Phe Asp Thr Arg Glu Pro Lys Ser Pro Leu
980 985 990
Glu Phe Ile Lys Asn Ser Asp Asp Val Ala Ala Tyr Asn Ile Ala Lys
995 1000 1005
Lys Arg Leu Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val
1010 1015 1020
<210> 81
<211> 1078
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.2-NLS融合蛋白的氨基酸序列
<400> 81
Met Lys Asn Gly Ile Asn Leu Phe Lys Thr Lys Thr Thr Lys Thr Lys
1 5 10 15
Gly Val Asp Met Glu Lys Tyr Gln Ile Thr Lys Thr Ile Arg Phe Lys
20 25 30
Leu Leu Pro Asp Asn Ala His Glu Ile Val Glu Lys Val Lys Ser Leu
35 40 45
Lys Thr Ser Asn Val Asp Glu Leu Met Asp Glu Val Lys Asn Val His
50 55 60
Leu Lys Gly Leu Glu Leu Leu Phe Ala Leu Lys Lys Tyr Phe Tyr Phe
65 70 75 80
Asp Gly Asn Gln Cys Lys Ser Phe Lys Ser Thr Leu Glu Ile Lys Ala
85 90 95
Arg Trp Leu Arg Leu Tyr Thr Pro Asp Gln Tyr Tyr Leu Lys Lys Ser
100 105 110
Ser Lys Asn Ser Tyr Gln Leu Lys Ser Leu Ser Tyr Phe Lys Asp Val
115 120 125
Phe Asn Asp Trp Leu Phe Asn Trp Glu Glu Ser Val Ser Glu Leu Ala
130 135 140
Ile Ile Tyr Glu Lys Tyr Lys Ile Cys Gln His Gln Arg Asp Ser Arg
145 150 155 160
Ala Asp Ile Ala Leu Leu Ile Lys Lys Leu Ser Met Lys Glu Tyr Phe
165 170 175
Pro Phe Ile Ser Asp Leu Ile Asp Cys Val Asn Asp Lys Asn Ser Asn
180 185 190
Lys Thr Phe Leu Met Lys Leu Ser Glu Glu Leu Ser Val Leu Leu Glu
195 200 205
Lys Cys Asn Ser Arg Ala Leu Pro Tyr Gln Ser Asn Gly Ile Val Val
210 215 220
Gly Lys Ala Ser Leu Asn Tyr Tyr Thr Val Ser Lys Ser Glu Lys Met
225 230 235 240
Leu Gln Asn Glu Tyr Glu Asp Val Cys Gln Ser Leu Asp Lys Asn Tyr
245 250 255
Asp Ile Thr Glu Met Lys Val Ile Leu Tyr Lys Glu Lys Leu Asp Asn
260 265 270
Leu Asn Phe Lys Asp Val Thr Ile Ala Asn Ala Tyr Asn Leu Leu Lys
275 280 285
Glu Asn Lys Ala Leu Gln Lys Arg Leu Phe Ser Glu Tyr Val Ser Gln
290 295 300
Gly Lys Val Leu Ser Leu Ile Lys Thr Glu Leu Pro Leu Phe Ser Asn
305 310 315 320
Ile Asn Asp Asn Asp Phe Glu Lys Tyr Lys Glu Trp Ser Asn Glu Ile
325 330 335
Lys Lys Leu Ala Asp Lys Lys Asn Thr Phe Cys Lys Lys Thr Gln Gln
340 345 350
Asp Lys Ile Lys Asp Ile Gln Asn Lys Ile Ser Glu Leu Lys Lys Lys
355 360 365
Arg Gly Ala Leu Phe Gln Tyr Lys Phe Thr Ser Phe Gln Lys His Cys
370 375 380
Asp Asn Tyr Lys Lys Val Ala Val Gln Tyr Gly Lys Leu Lys Ala Arg
385 390 395 400
Lys Lys Ala Ile Glu Lys Asp Glu Ile Glu Ala Asn Leu Leu Arg Tyr
405 410 415
Trp Ser Val Ile Leu Glu Gln Glu Asp Lys His Ser Leu Val Leu Ile
420 425 430
Pro Lys Asn Asn Ala Lys Asp Ala Lys Gln Tyr Ile Glu Thr Ile Asn
435 440 445
Thr Lys Gly Gly Lys Tyr Ile Ile His His Leu Asp Ser Leu Thr Leu
450 455 460
Arg Ala Leu Asn Lys Leu Cys Phe Asn Ala Val Asp Ile Glu Lys Gly
465 470 475 480
Gln Met Val Arg Glu Asn Thr Phe Tyr Gln Gly Ile Lys Glu Glu Phe
485 490 495
Glu Arg Asn Lys Ile Asn Cys Asp Asn Gln Gly Val Leu Lys Ile Gln
500 505 510
Gly Leu Tyr Ser Phe Lys Thr Glu Gly Gly Gln Ile Asn Glu Lys Glu
515 520 525
Ala Val Glu Phe Phe Lys Glu Val Leu Lys Ser Asn Tyr Ala Arg Glu
530 535 540
Val Leu Asn Leu Pro Tyr Asp Leu Glu Ser Asn Ile Phe Gln Lys Glu
545 550 555 560
Tyr Thr Asn Leu Asp Gln Phe Arg Gln Asp Leu Glu Lys Cys Cys Tyr
565 570 575
Ala Leu His Ser Lys Ile Gly Lys Asp Asp Leu Asp Glu Phe Thr Arg
580 585 590
Arg Phe Glu Ala Gln Val Phe Asp Ile Thr Ser Ile Asp Leu Lys Ser
595 600 605
Lys Lys Glu Lys Thr Lys Thr Thr Gly Glu Met Lys Lys His Thr Gln
610 615 620
Leu Trp Leu Glu Phe Trp Lys Gly Ala Ile Glu Gln Asn Phe Ala Thr
625 630 635 640
Arg Val Asn Pro Glu Leu Ser Ile Phe Trp Arg Ala Pro Lys Ser Ser
645 650 655
Arg Glu Lys Lys Tyr Gly Lys Gly Ser Asp Leu Tyr Asp Pro Asn Lys
660 665 670
Asn Asn Arg Tyr Leu Tyr Glu Gln Tyr Thr Leu Ala Leu Thr Ile Thr
675 680 685
Glu Asn Ala Gly Ser His Phe Lys Asp Ile Ala Phe Lys Asp Thr Ser
690 695 700
Lys Ile Lys Glu Ala Ile Lys Glu Phe Asn Met Ser Leu Ser Gln Ser
705 710 715 720
Lys Tyr Cys Phe Gly Ile Asp Arg Gly Asn Ala Glu Leu Val Ser Leu
725 730 735
Cys Leu Ile Lys Asn Glu Lys Asp Phe Pro Phe Glu Lys Phe Pro Val
740 745 750
Tyr Arg Leu Arg Asp Leu Thr Tyr Gln Gly Asp Phe Lys Asp Lys His
755 760 765
Asp Gln Met Arg Tyr Gly Val Ala Ile Lys Asn Ile Ser Tyr Phe Ile
770 775 780
Asp Gln Glu Asp Leu Phe Glu Lys Asn Asn Leu Ser Ala Ile Asp Met
785 790 795 800
Thr Thr Ala Lys Leu Ile Lys Asn Lys Ile Val Leu Asn Gly Asp Val
805 810 815
Leu Thr Tyr Leu Lys Leu Lys Glu Glu Thr Ala Lys His Lys Leu Thr
820 825 830
Gln Phe Phe Gln Gly Ser Ser Ile Asn Lys Asn Ser Arg Val Tyr Phe
835 840 845
Asp Glu Asp Glu Asn Val Phe Lys Ile Thr Thr Asn Arg Asn His Asn
850 855 860
Pro Glu Glu Ile Ile Tyr Phe Tyr Arg Gly Glu Tyr Gly Ala Ile Lys
865 870 875 880
Asn Lys Asn Asp Leu Glu Asp Ile Leu Asn Glu Tyr Leu Cys Lys Met
885 890 895
Glu Thr Gly Glu Ser Glu Ile Val Leu Leu Asn Arg Val Asn His Leu
900 905 910
Arg Asp Ala Ile Ser Ala Asn Ile Val Gly Ile Leu Ser Tyr Leu Ile
915 920 925
Asp Leu Phe Pro Glu Thr Ile Val Ala Leu Glu Asn Leu Ala Lys Gly
930 935 940
Thr Ile Asp Arg His Val Ser Gln Ser Tyr Glu Asn Ile Thr Arg Arg
945 950 955 960
Phe Glu Trp Ala Leu Tyr Arg Lys Leu Leu Asn Lys Gln Leu Ala Pro
965 970 975
Pro Glu Leu Lys Glu Asn Ile Leu Leu Arg Glu Gly Asp Asp Lys Ile
980 985 990
Asp Gln Phe Gly Ile Ile His Phe Val Glu Glu Lys Asn Thr Ser Lys
995 1000 1005
Asp Cys Pro Asn Cys Arg Lys Thr Thr Gln Gln Thr Asn Asp Asn
1010 1015 1020
Lys Phe Lys Glu Lys Lys Phe Val Cys Lys Ser Cys Gly Phe Asp
1025 1030 1035
Thr Ser Lys Asp Arg Lys Gly Met Asp Ser Leu Asn Ser Pro Asp
1040 1045 1050
Thr Val Ala Ala Tyr Asn Val Ala Arg Lys Lys Phe Glu Ser Ser
1055 1060 1065
Arg Ala Asp Pro Lys Lys Lys Arg Lys Val
1070 1075
<210> 82
<211> 1102
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.3-NLS融合蛋白的氨基酸序列
<400> 82
Met Ala Gly Thr Pro Tyr Thr Gly His Val Ala Cys Lys Tyr Cys Lys
1 5 10 15
Ile Thr Ser Trp Ala Thr Tyr Asp Arg Ile Lys Ile Asn Lys Ile Asn
20 25 30
Met Asn Gln Ser Phe Ile Asn Gly Gln Asn Phe Tyr Glu Leu Arg Lys
35 40 45
Thr Ile Arg Phe Val Leu Asp Pro Lys Thr Leu Lys Arg Pro Tyr Thr
50 55 60
Pro Ser Ser Asp Glu Val Asn Leu Glu Glu Gln Leu Asn Asn Phe Ile
65 70 75 80
Glu Lys Tyr Gln Gln Gly Ile Asn Asp Phe Lys Tyr Ile Val Tyr Phe
85 90 95
Gly Pro Lys Thr Ala Glu Thr Lys Glu Leu Asn Lys Lys Ile Ser Ile
100 105 110
Lys His Ser Trp Leu Arg Asn Tyr Thr Lys Ser Glu Phe Tyr Ser Ile
115 120 125
Lys Asp Lys Leu Ile Gln Leu Asp Tyr Asn Gly Asn Lys Ala Ser Ile
130 135 140
Gly Asn Ser Asn Leu Lys Phe Leu Asn Glu Tyr Phe Glu Asn Trp Ile
145 150 155 160
Ser Glu Asn Gln Glu Cys Ala Asp Ala Leu Lys Asn Cys Ile Asn Ala
165 170 175
Pro Ala Glu Lys Gln Lys Arg Lys Ser Glu Ala Ala His Trp Val Arg
180 185 190
Lys Leu Thr Lys Arg Ser Asn Phe Glu Cys Ile Phe Glu Leu Phe Asn
195 200 205
Gly Asn Ile Asp His Lys Asn Ser Asn Asp Asp Ile Glu Lys Ile Lys
210 215 220
His Cys Leu Asn Glu Cys Lys Thr Leu Leu Thr Ser Leu Glu Lys Met
225 230 235 240
Leu Leu Pro Ser Gln Ser Leu Gly Met Glu Ile Glu Arg Ala Ser Leu
245 250 255
Asn Tyr Tyr Thr Ile Asn Lys Lys Pro Lys Asn Tyr Asp Glu Asp Ile
260 265 270
Ala Gln Lys Ala Ser Ala Leu Asn Glu Ala Tyr Gln Phe Lys Ala Asp
275 280 285
Asp Lys Ala Phe Leu Asn Arg Val Gly Phe Ser Asp Asp Gly Val Pro
290 295 300
Ile Asn Glu Leu Lys Glu Ala Met Lys Lys Phe Lys Ala Asp Gln Lys
305 310 315 320
Ser Lys Phe Tyr Glu Phe Val Asn Gln Lys Lys Ser Tyr Ser Asp Leu
325 330 335
Lys Lys Asn Asp Asp Leu Lys Leu Leu Asn Asp Ile Ser Glu Glu Asp
340 345 350
Phe Asn Lys Phe Lys Glu Thr Gln Asp Lys Met Thr Arg Gly Lys His
355 360 365
Phe Gln Phe Ser Phe Pro Asn Tyr Lys Lys Ser Glu Lys Asn Phe Cys
370 375 380
Asp Leu Tyr Lys Asn Val Ala Val Ala Phe Gly Lys Ile Arg Ala Asp
385 390 395 400
Ile Lys Ala Leu Glu Lys Glu Arg Met Asp Ala Glu Lys Leu Gln Cys
405 410 415
Trp Ala Val Ile Leu Glu Lys Asp Asn Gln Arg Tyr Val Val Thr Ile
420 425 430
Pro Arg Asp Ala Asn Asn Asn Leu Thr Asn Thr Lys Gln Tyr Ile Asp
435 440 445
Asn Leu Gln Asn Glu Glu Asn Asp Gln Trp Ile Leu Tyr Ala Phe Glu
450 455 460
Ser Leu Thr Leu Arg Ser Leu Asp Lys Leu Cys Phe Gly Leu Asp Lys
465 470 475 480
Asn Thr Phe Ile Pro Ala Ile Thr Gly Glu Leu Tyr Gln Lys Asn Asn
485 490 495
Ser Phe Phe Glu Lys Gly Leu Leu Lys Arg Lys Asp Gln Phe Ser Gln
500 505 510
Asn Gly Thr Asp Leu Ala Ala Phe Tyr Lys Thr Val Leu Glu Leu Asp
515 520 525
Ser Thr Lys Lys Met Leu Gly Ile Asn Lys Tyr Ala Asp Phe Lys Ala
530 535 540
Phe Ile Ser Lys Glu Tyr Thr Ala Leu Glu Asp Phe Glu Lys Thr Leu
545 550 555 560
Lys Glu Thr Cys Tyr Phe Lys Lys Arg Val Phe Ile Ser Glu Asp Thr
565 570 575
Lys Asn Lys Leu Ile Asn Asp Tyr Gln Gly Asn Leu Tyr Lys Ile Thr
580 585 590
Ser Tyr Asp Leu Glu Lys Asp Asp Ser Glu Ala Leu Gly Thr Leu Ile
595 600 605
Asn Lys Lys Gln Phe Asn Arg Ala Ser Pro Glu Ile His Thr Lys Thr
610 615 620
Trp Leu Asp Phe Trp Thr Ala Asp Asn Glu Thr Asp Lys Tyr Pro Ile
625 630 635 640
Arg Leu Asn Pro Glu Phe Lys Ile Ser Phe Val Glu Lys Gln Asp Lys
645 650 655
Asp Leu Asn Met Arg Asn Leu Gly Leu Leu Asn Lys Asn Arg Arg Leu
660 665 670
Lys Ser Gln Phe Leu Leu Ser Thr Thr Ile Thr Leu Leu Ala His Glu
675 680 685
Lys Asn Ala Asp Leu His Phe Lys Lys Thr Asp Glu Ile Gln Thr Phe
690 695 700
Ile Asn Ser Tyr Asn Gln Glu Phe Asn Lys Lys Ile Lys Pro Phe Asp
705 710 715 720
Ile Tyr Tyr Tyr Gly Leu Asp Arg Gly Gln Lys Glu Leu Leu Thr Leu
725 730 735
Gly Leu Phe Lys Phe Ser Glu Asn Glu Lys Val Ser Phe Thr Lys Gln
740 745 750
Asp Gly Thr Val Gly Glu Tyr Ser Lys Pro Lys Phe Ile Pro Leu Asp
755 760 765
Val Tyr Gln Ile Arg Glu Gly Gln Tyr Leu Thr Lys Asn Lys Lys Gly
770 775 780
Arg Leu Ala Tyr Lys Ser Ile Asp Gln Phe Ile Asp Asp Glu Lys Val
785 790 795 800
Ile Glu Lys Leu Pro Val Asn Ser Cys Leu Asp Leu Ser Cys Ala Lys
805 810 815
Leu Val Lys Gly Lys Ile Ile Gln Asn Gly Asp Val Ala Thr Tyr Leu
820 825 830
Glu Leu Lys Arg Val Ser Ala Leu Arg Lys Ile Tyr Glu Asn Thr Thr
835 840 845
Arg Gly Gln Phe Lys Thr Asp Arg Ile Gly Phe Asn Lys Asp Lys Gly
850 855 860
Cys Leu Phe Leu Asp Ile Glu Asn Arg Gly Lys Leu Glu Asn Asn Asn
865 870 875 880
Leu Tyr Phe Tyr Asp Asn Arg Phe Ala Glu Ile Leu Ser Leu Asp Ser
885 890 895
Ile Ile Lys Glu Leu Gln Asp Tyr Tyr Asn Glu Val Lys Asn Lys Gln
900 905 910
Asn Ile Glu Phe Ile Ser Ile Asp Lys Ile Asn His Leu Arg Asp Ala
915 920 925
Leu Cys Ala Asn Ala Val Gly Ile Leu Ala His Leu Gln Lys Thr His
930 935 940
Phe Gly Val Ile Val Phe Glu Gly Leu Asp Ala Arg His Lys Asn Lys
945 950 955 960
Glu Thr Thr Glu Phe Ala Gly Asn Leu Ala Ser Arg Ile Glu Arg Lys
965 970 975
Ile Leu Gln Lys Leu Glu Thr Leu Ser Leu Ile Pro Pro Gln His Arg
980 985 990
Gln Ile Ile Asp Leu Gln Asn Ser Lys Gln Ile Lys Gln Thr Gly Ala
995 1000 1005
Val Leu Tyr Ile Glu Glu Lys Gly Thr Ser Ala Asn Cys Pro His
1010 1015 1020
Cys Glu Thr Ala Asn Pro Asp Lys Ser Glu Lys Trp Leu Ala His
1025 1030 1035
Asn Tyr Lys Cys Lys Asn Ser Asn Cys Asn Phe Asp Ala Ser Glu
1040 1045 1050
Ile Ser Lys Arg Lys Asp Leu Ile Gly Leu Asp Asn Ser Asp Ser
1055 1060 1065
Val Ala Thr Tyr Asn Ile Ala Lys Arg Gly Leu Leu Glu Met Asn
1070 1075 1080
Gln Lys Ile Glu Gln Ser Lys Val Ser Arg Ala Asp Pro Lys Lys
1085 1090 1095
Lys Arg Lys Val
1100
<210> 83
<211> 1068
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.4-NLS融合蛋白的氨基酸序列
<400> 83
Met Glu Ile Gln Glu Leu Lys Asn Leu Tyr Glu Val Lys Lys Thr Val
1 5 10 15
Arg Phe Glu Leu Lys Pro Ser Lys Lys Lys Ile Phe Glu Gly Gly Asp
20 25 30
Val Ile Lys Leu Gln Lys Asp Phe Glu Lys Val Gln Lys Phe Phe Leu
35 40 45
Asp Ile Phe Val Tyr Lys Asn Glu His Thr Lys Leu Glu Phe Lys Lys
50 55 60
Lys Arg Glu Ile Lys Tyr Thr Trp Leu Arg Thr Asn Thr Lys Asn Glu
65 70 75 80
Phe Tyr Asn Trp Arg Gly Lys Ser Asp Thr Gly Lys Asn Tyr Ala Leu
85 90 95
Asn Lys Ile Gly Phe Leu Ala Glu Glu Ile Leu Arg Trp Leu Asn Glu
100 105 110
Trp Gln Glu Leu Thr Lys Ser Leu Lys Asp Leu Thr Gln Arg Glu Glu
115 120 125
His Lys Gln Glu Arg Lys Ser Asp Ile Ala Phe Val Leu Arg Asn Phe
130 135 140
Leu Lys Arg Gln Asn Leu Pro Phe Ile Lys Asp Phe Phe Asn Ala Val
145 150 155 160
Ile Asp Ile Gln Gly Lys Gln Gly Lys Glu Ser Asp Asp Lys Ile Arg
165 170 175
Lys Phe Arg Glu Glu Ile Lys Glu Ile Glu Lys Asn Leu Asn Ala Cys
180 185 190
Ser Arg Glu Tyr Leu Pro Thr Gln Ser Asn Gly Val Leu Leu Tyr Lys
195 200 205
Ala Ser Phe Ser Tyr Tyr Thr Leu Asn Lys Thr Pro Lys Glu Tyr Glu
210 215 220
Asp Leu Lys Lys Glu Lys Glu Ser Glu Leu Ser Ser Val Leu Leu Lys
225 230 235 240
Glu Ile Tyr Arg Arg Lys Arg Phe Asn Arg Thr Thr Asn Gln Lys Asp
245 250 255
Thr Leu Phe Glu Cys Thr Ser Asp Trp Leu Val Lys Ile Lys Leu Gly
260 265 270
Lys Asp Ile Tyr Glu Trp Thr Leu Asp Glu Ala Tyr Gln Lys Met Lys
275 280 285
Ile Trp Lys Ala Asn Gln Lys Ser Asn Phe Ile Glu Ala Val Ala Gly
290 295 300
Asp Lys Leu Thr His Gln Asn Phe Arg Lys Gln Phe Pro Leu Phe Asp
305 310 315 320
Ala Ser Asp Glu Asp Phe Glu Thr Phe Tyr Arg Leu Thr Lys Ala Leu
325 330 335
Asp Lys Asn Pro Glu Asn Ala Lys Lys Ile Ala Gln Lys Arg Gly Lys
340 345 350
Phe Phe Asn Ala Pro Asn Glu Thr Val Gln Thr Lys Asn Tyr His Glu
355 360 365
Leu Cys Glu Leu Tyr Lys Arg Ile Ala Val Lys Arg Gly Lys Ile Ile
370 375 380
Ala Glu Ile Lys Gly Ile Glu Asn Glu Glu Val Gln Ser Gln Leu Leu
385 390 395 400
Thr His Trp Ala Val Ile Ala Glu Glu Arg Asp Lys Lys Phe Ile Val
405 410 415
Leu Ile Pro Arg Lys Asn Gly Gly Lys Leu Glu Asn His Lys Asn Ala
420 425 430
His Ala Phe Leu Gln Glu Lys Asp Arg Lys Glu Pro Asn Asp Ile Lys
435 440 445
Val Tyr His Phe Lys Ser Leu Thr Leu Arg Ser Leu Glu Lys Leu Cys
450 455 460
Phe Lys Glu Ala Lys Asn Thr Phe Ala Pro Glu Ile Lys Lys Glu Thr
465 470 475 480
Asn Pro Lys Ile Trp Phe Pro Thr Tyr Lys Gln Glu Trp Asn Ser Thr
485 490 495
Pro Glu Arg Leu Ile Lys Phe Tyr Lys Gln Val Leu Gln Ser Asn Tyr
500 505 510
Ala Gln Thr Tyr Leu Asp Leu Val Asp Phe Gly Asn Leu Asn Thr Phe
515 520 525
Leu Glu Thr His Phe Thr Thr Leu Glu Glu Phe Glu Ser Asp Leu Glu
530 535 540
Lys Thr Cys Tyr Thr Lys Val Pro Val Tyr Phe Ala Lys Lys Glu Leu
545 550 555 560
Glu Thr Phe Ala Asp Glu Phe Glu Ala Glu Val Phe Glu Ile Thr Thr
565 570 575
Arg Ser Ile Ser Thr Glu Ser Lys Arg Lys Glu Asn Ala His Ala Glu
580 585 590
Ile Trp Arg Asp Phe Trp Ser Arg Glu Asn Glu Glu Glu Asn His Ile
595 600 605
Thr Arg Leu Asn Pro Glu Val Ser Val Leu Tyr Arg Asp Glu Ile Lys
610 615 620
Glu Lys Ser Asn Thr Ser Arg Lys Asn Arg Lys Ser Asn Ala Asn Asn
625 630 635 640
Arg Phe Ser Asp Pro Arg Phe Thr Leu Ala Thr Thr Ile Thr Leu Asn
645 650 655
Ala Asp Lys Lys Lys Ser Asn Leu Ala Phe Lys Thr Val Glu Asp Ile
660 665 670
Asn Ile His Ile Asp Asn Phe Asn Lys Lys Phe Ser Lys Asn Phe Ser
675 680 685
Gly Glu Trp Val Tyr Gly Ile Asp Arg Gly Leu Lys Glu Leu Ala Thr
690 695 700
Leu Asn Val Val Lys Phe Ser Asp Val Lys Asn Val Phe Gly Val Ser
705 710 715 720
Gln Pro Lys Glu Phe Ala Lys Ile Pro Ile Tyr Lys Leu Arg Asp Glu
725 730 735
Lys Ala Ile Leu Lys Asp Glu Asn Gly Leu Ser Leu Lys Asn Ala Lys
740 745 750
Gly Glu Ala Arg Lys Val Ile Asp Asn Ile Ser Asp Val Leu Glu Glu
755 760 765
Gly Lys Glu Pro Asp Ser Thr Leu Phe Glu Lys Arg Glu Val Ser Ser
770 775 780
Ile Asp Leu Thr Arg Ala Lys Leu Ile Lys Gly His Ile Ile Ser Asn
785 790 795 800
Gly Asp Gln Lys Thr Tyr Leu Lys Leu Lys Glu Thr Ser Ala Lys Arg
805 810 815
Arg Ile Phe Glu Leu Phe Ser Thr Ala Lys Ile Asp Lys Ser Ser Gln
820 825 830
Phe His Val Arg Lys Thr Ile Glu Leu Ser Gly Thr Lys Ile Tyr Trp
835 840 845
Leu Cys Glu Trp Gln Arg Gln Asp Ser Trp Arg Thr Glu Lys Val Ser
850 855 860
Leu Arg Asn Thr Leu Lys Gly Tyr Leu Gln Asn Leu Asp Leu Lys Asn
865 870 875 880
Arg Phe Glu Asn Ile Glu Thr Ile Glu Lys Ile Asn His Leu Arg Asp
885 890 895
Ala Ile Thr Ala Asn Met Val Gly Ile Leu Ser His Leu Gln Asn Lys
900 905 910
Leu Glu Met Gln Gly Val Ile Ala Leu Glu Asn Leu Asp Thr Val Arg
915 920 925
Glu Gln Ser Asn Lys Lys Met Ile Asp Glu His Phe Glu Gln Ser Asn
930 935 940
Glu His Val Ser Arg Arg Leu Glu Trp Ala Leu Tyr Cys Lys Phe Ala
945 950 955 960
Asn Thr Gly Glu Val Pro Pro Gln Ile Lys Glu Ser Ile Phe Leu Arg
965 970 975
Asp Glu Phe Lys Val Cys Gln Ile Gly Ile Leu Asn Phe Ile Asp Val
980 985 990
Lys Gly Thr Ser Ser Asn Cys Pro Asn Cys Asp Gln Glu Ser Arg Lys
995 1000 1005
Thr Gly Ser His Phe Ile Cys Asn Phe Gln Asn Asn Cys Ile Phe
1010 1015 1020
Ser Ser Lys Glu Asn Arg Asn Leu Leu Glu Gln Asn Leu His Asn
1025 1030 1035
Ser Asp Asp Val Ala Ala Phe Asn Ile Ala Lys Arg Gly Leu Glu
1040 1045 1050
Ile Val Lys Val Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val
1055 1060 1065
<210> 84
<211> 1169
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.5-NLS融合蛋白的氨基酸序列
<400> 84
Met Glu Asn Phe Lys Asn Leu Tyr Glu Val Arg Lys Thr Val Arg Phe
1 5 10 15
Glu Leu Lys Pro Ser Arg Lys Lys Thr Phe Ala Gly Gly Asp Ile Phe
20 25 30
Glu Leu Gln Lys Asp Phe Glu Glu Val Gln Lys Phe Phe Leu Asp Ile
35 40 45
Phe Val Phe Ala Ile Glu Gln Glu Lys Leu Tyr Gln Glu Glu Glu Glu
50 55 60
Glu Gly Lys Leu Ser Arg Tyr Thr Lys Ile Glu Phe Lys Lys Lys Arg
65 70 75 80
Glu Ile Lys Tyr Thr Trp Leu Arg Ile Tyr Thr Lys Asn Glu Phe Tyr
85 90 95
Asp Trp Asn Gly Lys Asn Asp Lys Glu Lys Asn Tyr Ala Leu Ser Lys
100 105 110
Ile Asp Phe Leu Glu Lys Glu Ile Leu Arg Trp Phe Asn Glu Trp Gln
115 120 125
Glu Leu Thr Val Asn Leu Lys Asn Leu Thr Gln Thr Lys Glu His Glu
130 135 140
Lys Glu Arg Lys Ser Asp Ile Ala Phe Val Leu Arg Asn Phe Leu Lys
145 150 155 160
Arg Gln Asn Phe Pro Phe Ile Lys Asp Phe Phe Asn Ala Val Ile Asp
165 170 175
Ile Gln Glu Lys Gln Gly Asn Glu Ser Asp Glu Lys Ile Arg Lys Phe
180 185 190
Arg Glu Glu Leu Arg Glu Met Lys Lys Asn Leu Asn Thr Cys Ala Lys
195 200 205
Glu Tyr Leu Ser Ser Gln Ser Lys Gly Val Leu Leu His Lys Ala Ser
210 215 220
Phe Asn Tyr Tyr Thr Leu Asn Lys Thr Pro Lys Glu Tyr Glu Asn Leu
225 230 235 240
Lys Leu Gln Lys Glu Leu Glu Ile Asp Asn Ile Leu Pro Lys Lys Ile
245 250 255
Cys Lys Arg Val Arg Trp Asn Lys Glu Lys Lys Gln Glu Asp Ile Leu
260 265 270
Phe Glu Cys Asn Ser Asp Trp Leu Val Glu Ile Lys Leu Gly Tyr Asp
275 280 285
Ile Gln Lys Trp Thr Leu Asp Glu Ala Tyr Gln Lys Met Lys Thr Trp
290 295 300
Lys Ala Asp Gln Lys Ser Asp Phe Asn Glu Lys Ile Gly Asn Phe Ile
305 310 315 320
Asp Gln Tyr Leu Lys Lys Gly Phe Ile Glu Asp Leu Met Asn Glu Asn
325 330 335
Glu Lys Lys Asn Ala Glu Ala Ile Leu Arg Glu Phe Ser Val Phe Lys
340 345 350
Pro Ile Glu Asn Phe Tyr Phe Tyr Asp Phe Leu Glu Arg Thr Lys Glu
355 360 365
Ile Lys Ile Leu Ser Asn Gln Lys Asn Asn Ile Leu Gln Lys Tyr Asn
370 375 380
Lys Asn Ala Lys Tyr Phe Glu Lys Ile Ile Thr Tyr Lys Ile Lys Asp
385 390 395 400
Lys Glu Asp Leu Thr Glu Asp Glu Lys Glu Tyr Gln Glu Leu Glu Lys
405 410 415
Ser Ile Glu Lys Lys Ala Lys Glu Arg Gly Lys Phe Phe Asn Ala Pro
420 425 430
Lys Glu Lys Val Gln Thr Gln His Tyr Phe Glu Leu Cys Glu Leu Tyr
435 440 445
Lys Arg Ile Ala Met Lys Arg Gly Lys Ile Ile Ala Glu Ile Lys Gly
450 455 460
Ile Glu Asn Glu Glu Val Gln Ser Gln Leu Leu Thr His Trp Ala Leu
465 470 475 480
Ile Ala Glu Glu Gly Glu Lys Lys Ser Val Val Phe Ile Pro Arg Lys
485 490 495
Asn Gly Glu Glu Leu Glu Asn His Lys Lys Ala His Glu Phe Leu Gln
500 505 510
Lys Gln Glu Lys Lys Glu Phe Gly Asp Ile Lys Ser Tyr His Phe Lys
515 520 525
Ser Leu Thr Leu Arg Ala Leu Glu Lys Leu Cys Phe Lys Glu Thr Glu
530 535 540
Asn Thr Phe Thr Pro Glu Ile Lys Lys Glu Thr Asn Pro Lys Val Trp
545 550 555 560
Phe Pro Lys Tyr Lys Gln Glu Trp Asn Asp Glu Pro Gln Lys Leu Ile
565 570 575
Asn Phe Tyr Lys Gln Val Leu Gln Ser Lys Tyr Ser Gln Lys Tyr Leu
580 585 590
Asp Leu Val Ala Phe Gly Asp Leu Lys Ser Phe Leu Glu Thr Ser Phe
595 600 605
Asp Asp Leu Gln Ile Phe Glu Ser Gly Leu Glu Lys Thr Cys Tyr Ile
610 615 620
Lys Val Pro Ile Tyr Phe Ser Lys Glu Gly Phe Glu Thr Phe Thr Asn
625 630 635 640
Arg Phe Asp Ala Glu Val Phe Glu Ile Thr Thr Arg Ser Ile Ser Ser
645 650 655
Glu Ser Lys Arg Lys Glu Asn Ala His Ala Glu Ile Trp Lys Asp Phe
660 665 670
Trp Ser Lys Glu Asn Glu Glu Lys Asn His Ile Thr Arg Leu Asn Pro
675 680 685
Glu Val Ser Val Phe Tyr Arg Asp Glu Ile Glu Lys Lys Ser Asn Ala
690 695 700
Leu Arg Gly Asn Asn Lys Ser Asn Ile Asn Asn Arg Phe Ser Ala Ser
705 710 715 720
Arg Phe Thr Leu Val Thr Thr Ile Thr Ile Arg Ala Thr His Lys Lys
725 730 735
Ser Asn Leu Ala Phe Lys Thr Glu Glu Asp Ile Lys Ser His Ile Asp
740 745 750
Lys Phe Asn Glu Ala Phe Gln Asn Phe Ser Gly Glu Trp Val Tyr Gly
755 760 765
Ile Asp Arg Gly Leu Lys Glu Leu Ala Thr Leu Asn Val Val Lys Phe
770 775 780
Ser Asp Glu Lys Asn Glu Phe Gly Val Ile Lys Pro Lys Glu Phe Ala
785 790 795 800
Lys Ile Pro Val Tyr Lys Leu Lys Asp Glu Lys Ala Ile Leu Lys Asp
805 810 815
Glu Asn Gly Lys Asp Leu Lys Asn Ala Lys Gly Glu Ala Arg Lys Val
820 825 830
Ile Asp Asn Ile Ser Glu Val Leu Glu Glu Lys Lys Glu Pro Asp Ser
835 840 845
Asn Leu Phe Glu Lys Gln Gly Val Leu Ser Gln Gly Ile Ser Cys Ile
850 855 860
Asp Leu Thr Gln Ala Lys Leu Ile Lys Gly His Ile Ile Leu Asn Gly
865 870 875 880
Asp Gln Lys Thr Tyr Leu Lys Leu Lys Glu Ile Ser Ala Lys Arg Arg
885 890 895
Ile Phe Glu Leu Phe Ser Thr Ser Lys Ile Asp Lys Asn Ser Glu Leu
900 905 910
Arg Val Glu Lys Thr Thr Ile Ser Ile Asn Ser Glu Asp Gly Lys Arg
915 920 925
Asp Phe Tyr Trp Leu Thr Lys Asn Gln Ile Val Asn Ser Glu Thr Lys
930 935 940
Lys Glu Ile Gln Lys Glu Gln Gln Glu Lys Leu Asp Asn Leu Lys Val
945 950 955 960
Ile Phe Ile Asp Tyr Leu Glu Gly Leu Cys Val Lys Asn Lys Phe Glu
965 970 975
Asp Ile Glu Thr Ile Glu Lys Ile Asn His Leu Arg Asp Ala Ile Thr
980 985 990
Ala Asn Met Val Gly Ile Leu Phe His Leu Gln Lys Glu Phe Lys Gly
995 1000 1005
Ile Ile Ala Leu Glu Asn Leu Asp Thr Val Arg Glu Gln Ser Asn
1010 1015 1020
Lys Lys Met Ile Asp Glu His Phe Glu Gln Ser Asn Glu Asp Ile
1025 1030 1035
Ser Arg Arg Leu Glu Trp Ala Leu Tyr Arg Lys Phe Ala Asn Met
1040 1045 1050
Gly Glu Val Pro Ser Gln Ile Lys Glu Ser Ile Phe Leu Arg Asp
1055 1060 1065
Glu Phe Lys Val Tyr Gln Met Gly Leu Leu Lys Phe Val Glu Val
1070 1075 1080
Ser Gly Thr Ser Ser Asn Cys Pro Asn Cys Asp Lys Glu Val Gly
1085 1090 1095
Lys Thr Asn Ser His Phe Val Cys Lys Gly Glu Asn Asn Cys Gly
1100 1105 1110
Phe Ser Ser Lys Glu Asn Arg Asn Leu Leu Glu Gln Asn Leu Asn
1115 1120 1125
Asn Ser Asp Glu Val Ala Ala Tyr Asn Ile Ala Lys Arg Gly Leu
1130 1135 1140
Lys Leu Ile Asn Gln Lys Trp Asn Asn Thr Ser Lys Ser Gln Asn
1145 1150 1155
Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val
1160 1165
<210> 85
<211> 1075
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.6-NLS融合蛋白的氨基酸序列
<400> 85
Met Glu Lys Tyr Lys Ile Thr Lys Thr Ile Arg Phe Lys Leu Leu Pro
1 5 10 15
Asp Lys Ile Gln Asp Ile Ser Arg Gln Val Ala Val Leu Gln Asn Ser
20 25 30
Thr Asn Ala Glu Lys Lys Asn Asn Leu Leu Arg Leu Val Gln Arg Gly
35 40 45
Gln Glu Leu Pro Lys Leu Leu Asn Glu Tyr Ile Arg Tyr Ser Asp Asn
50 55 60
His Lys Leu Lys Ser Asn Val Thr Val His Phe Arg Trp Leu Arg Leu
65 70 75 80
Phe Thr Lys Asp Leu Phe Tyr Asn Trp Lys Lys Asp Asn Thr Glu Lys
85 90 95
Lys Ile Lys Ile Ser Asp Val Val Tyr Leu Ser His Val Phe Glu Ala
100 105 110
Phe Leu Lys Glu Trp Glu Ser Thr Ile Glu Arg Val Asn Ala Asp Cys
115 120 125
Asn Lys Pro Glu Glu Ser Lys Thr Arg Asp Ala Glu Ile Ala Leu Ser
130 135 140
Ile Arg Lys Leu Gly Ile Lys His Gln Leu Pro Phe Ile Lys Gly Phe
145 150 155 160
Val Asp Asn Ser Asn Asp Lys Asn Ser Glu Asp Thr Lys Ser Lys Leu
165 170 175
Thr Ala Leu Leu Ser Glu Phe Glu Ala Val Leu Lys Ile Cys Glu Gln
180 185 190
Asn Tyr Leu Pro Ser Gln Ser Ser Gly Ile Ala Ile Ala Lys Ala Ser
195 200 205
Phe Asn Tyr Tyr Thr Ile Asn Lys Lys Gln Lys Asp Phe Glu Ala Glu
210 215 220
Ile Val Ala Leu Lys Lys Gln Leu His Ala Arg Tyr Gly Asn Lys Lys
225 230 235 240
Tyr Asp Gln Leu Leu Arg Glu Leu Asn Leu Ile Pro Leu Lys Glu Leu
245 250 255
Pro Leu Lys Glu Leu Pro Leu Ile Glu Phe Tyr Ser Glu Ile Lys Lys
260 265 270
Arg Lys Ser Thr Lys Lys Ser Glu Phe Leu Glu Ala Val Ser Asn Gly
275 280 285
Leu Val Phe Asp Asp Leu Lys Ser Lys Phe Pro Leu Phe Gln Thr Glu
290 295 300
Ser Asn Lys Tyr Asp Glu Tyr Leu Lys Leu Ser Asn Lys Ile Thr Gln
305 310 315 320
Lys Ser Thr Ala Lys Ser Leu Leu Ser Lys Asp Ser Pro Glu Ala Gln
325 330 335
Lys Leu Gln Thr Glu Ile Thr Lys Leu Lys Lys Asn Arg Gly Glu Tyr
340 345 350
Phe Lys Lys Ala Phe Gly Lys Tyr Val Gln Leu Cys Glu Leu Tyr Lys
355 360 365
Glu Ile Ala Gly Lys Arg Gly Lys Leu Lys Gly Gln Ile Lys Gly Ile
370 375 380
Glu Asn Glu Arg Ile Asp Ser Gln Arg Leu Gln Tyr Trp Ala Leu Val
385 390 395 400
Leu Glu Asp Asn Leu Lys His Ser Leu Ile Leu Ile Pro Lys Glu Lys
405 410 415
Thr Asn Glu Leu Tyr Arg Lys Val Trp Gly Ala Lys Asp Asp Gly Ala
420 425 430
Ser Ser Ser Ser Ser Ser Thr Leu Tyr Tyr Phe Glu Ser Met Thr Tyr
435 440 445
Arg Ala Leu Arg Lys Leu Cys Phe Gly Ile Asn Gly Asn Thr Phe Leu
450 455 460
Pro Glu Ile Gln Lys Glu Leu Pro Gln Tyr Asn Gln Lys Glu Phe Gly
465 470 475 480
Glu Phe Cys Phe His Lys Ser Asn Asp Asp Lys Glu Ile Asp Glu Pro
485 490 495
Lys Leu Ile Ser Phe Tyr Gln Ser Val Leu Lys Thr Asp Phe Val Lys
500 505 510
Asn Thr Leu Ala Leu Pro Gln Ser Val Phe Asn Glu Val Ala Ile Gln
515 520 525
Ser Phe Glu Thr Arg Gln Asp Phe Gln Ile Ala Leu Glu Lys Cys Cys
530 535 540
Tyr Ala Lys Lys Gln Ile Ile Ser Glu Ser Leu Lys Lys Glu Ile Leu
545 550 555 560
Glu Asn Tyr Asn Thr Gln Ile Phe Lys Ile Thr Ser Leu Asp Leu Gln
565 570 575
Arg Ser Glu Gln Lys Asn Leu Lys Gly His Thr Arg Ile Trp Asn Arg
580 585 590
Phe Trp Thr Lys Gln Asn Glu Glu Ile Asn Tyr Asn Leu Arg Leu Asn
595 600 605
Pro Glu Ile Ala Ile Val Trp Arg Lys Ala Lys Lys Thr Arg Ile Glu
610 615 620
Lys Tyr Gly Glu Arg Ser Val Leu Tyr Glu Pro Glu Lys Arg Asn Arg
625 630 635 640
Tyr Leu His Glu Gln Tyr Thr Leu Cys Thr Thr Val Thr Asp Asn Ala
645 650 655
Leu Asn Asn Glu Ile Thr Phe Ala Phe Glu Asp Thr Lys Lys Lys Gly
660 665 670
Thr Glu Ile Val Lys Tyr Asn Glu Lys Ile Asn Gln Thr Leu Lys Lys
675 680 685
Glu Phe Asn Lys Asn Gln Leu Trp Phe Tyr Gly Ile Asp Ala Gly Glu
690 695 700
Ile Glu Leu Ala Thr Leu Ala Leu Met Asn Lys Asp Lys Glu Pro Gln
705 710 715 720
Leu Phe Thr Val Tyr Glu Leu Lys Lys Leu Asp Phe Phe Lys His Gly
725 730 735
Tyr Ile Tyr Asn Lys Glu Arg Glu Leu Val Ile Arg Glu Lys Pro Tyr
740 745 750
Lys Ala Ile Gln Asn Leu Ser Tyr Phe Leu Asn Glu Glu Leu Tyr Glu
755 760 765
Lys Thr Phe Arg Asp Gly Lys Phe Asn Glu Thr Tyr Asn Glu Leu Phe
770 775 780
Lys Glu Lys His Val Ser Ala Ile Asp Leu Thr Thr Ala Lys Val Ile
785 790 795 800
Asn Gly Lys Ile Ile Leu Asn Gly Asp Met Ile Thr Phe Leu Asn Leu
805 810 815
Arg Ile Leu His Ala Gln Arg Lys Ile Tyr Glu Glu Leu Ile Glu Asn
820 825 830
Pro His Ala Glu Leu Lys Glu Lys Asp Tyr Lys Leu Tyr Phe Glu Ile
835 840 845
Glu Gly Lys Asp Lys Asp Ile Tyr Ile Ser Arg Leu Asp Phe Glu Tyr
850 855 860
Ile Lys Pro Tyr Gln Glu Ile Ser Asn Tyr Leu Phe Ala Tyr Phe Ala
865 870 875 880
Ser Gln Gln Ile Asn Glu Ala Arg Glu Glu Glu Gln Ile Asn Gln Thr
885 890 895
Lys Arg Ala Leu Ala Gly Asn Met Ile Gly Val Ile Tyr Tyr Leu Tyr
900 905 910
Gln Lys Tyr Arg Gly Ile Ile Ser Ile Glu Asp Leu Lys Gln Thr Lys
915 920 925
Val Glu Ser Asp Arg Asn Lys Phe Glu Gly Asn Ile Glu Arg Pro Leu
930 935 940
Glu Trp Ala Leu Tyr Arg Lys Phe Gln Gln Glu Gly Tyr Val Pro Pro
945 950 955 960
Ile Ser Glu Leu Ile Lys Leu Arg Glu Leu Glu Lys Phe Pro Leu Lys
965 970 975
Asp Val Lys Gln Pro Lys Tyr Glu Asn Ile Gln Gln Phe Gly Ile Ile
980 985 990
Lys Phe Val Ser Pro Glu Glu Thr Ser Thr Thr Cys Pro Lys Cys Leu
995 1000 1005
Arg Arg Phe Lys Asp Tyr Asp Lys Asn Lys Gln Glu Gly Phe Cys
1010 1015 1020
Lys Cys Gln Cys Gly Phe Asp Thr Arg Asn Asp Leu Lys Gly Phe
1025 1030 1035
Glu Gly Leu Asn Asp Pro Asp Lys Val Ala Ala Phe Asn Ile Ala
1040 1045 1050
Lys Arg Gly Phe Glu Asp Leu Gln Lys Tyr Lys Ser Arg Ala Asp
1055 1060 1065
Pro Lys Lys Lys Arg Lys Val
1070 1075
<210> 86
<211> 1117
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.7-NLS融合蛋白的氨基酸序列
<400> 86
Met Leu Ile Gln Phe Lys Asn His Tyr Ser Tyr Asn Lys Ser Ile Arg
1 5 10 15
Phe Lys Leu Glu His Lys Asn Gly Lys Leu Pro Lys Leu Glu Ser Asp
20 25 30
Asn Val Asp Leu Asn Lys Leu Val Asp Ile Gly Asn Ser Leu Lys Asp
35 40 45
Ile Phe Glu Glu Leu Val Tyr Thr Lys Asn Asn Tyr Asn Lys Leu Asn
50 55 60
Ser Leu Val Ser Ile Lys Lys Gln Trp Leu Lys Ile Tyr Phe Lys Asn
65 70 75 80
Glu Phe Tyr Ser Asn Gly Lys Ile Gln Asn Tyr Ser Leu Ser Asn Phe
85 90 95
Ser Tyr Leu Pro Asn Lys Leu Ile Glu Trp Leu Asn Asn Trp Gln Asn
100 105 110
Asn Leu Lys Ala Leu Ile Glu Leu Thr Lys Gln Gln Asp Phe Asn Lys
115 120 125
Thr Lys Lys Ser Glu Ile Ala Tyr Ile Leu Ser Leu Phe Asn Gly Lys
130 135 140
Tyr Ser Phe Ser Phe Val Lys Asp Phe Ser Thr Cys Ile Asn His Lys
145 150 155 160
Asn Ser Gln Glu Gln Ile Leu Lys Leu Gln Gly Val Val Glu Asn Phe
165 170 175
Glu Lys Val Leu Asn Leu Cys Ile Gln Glu Tyr Leu Pro Ser Lys Ser
180 185 190
Ala Gly Val Val Ile Ala Gln Gly Ser Met Asn Tyr Tyr Ala Ile Asn
195 200 205
Lys Glu Pro Lys Arg Tyr Asp Asn Ile Leu Ala Asp Leu Asn Gln Lys
210 215 220
Phe Glu Glu Leu Asp Lys Glu Tyr Ile Ala Met Lys Gln Tyr Lys Ser
225 230 235 240
Ser Gln Lys Ser Arg Leu Phe Glu Phe Ile Arg Lys Gly Phe Ser Lys
245 250 255
Asp Gln Ile Leu Ser Glu Phe Lys Lys Lys Glu Asn Asn Glu Val Ser
260 265 270
Phe Val Tyr Asn Asn Gln Ile Ile Ile Arg Ile Tyr Thr Gln Glu Leu
275 280 285
Phe Lys Asp Ser Tyr Cys Leu Gly Glu Val Ile Lys Leu Thr Lys Lys
290 295 300
Ile Glu Glu Leu Asn Glu Ser Lys Asp Ser Asn Asn Asn Leu Pro Glu
305 310 315 320
Glu Thr Lys Lys Glu Ile Thr Lys Leu Lys Lys Glu Ile Gly Phe Tyr
325 330 335
Phe Ile Arg Arg Thr Arg Gly Lys Ser His Asn Asn Tyr Phe Lys Ser
340 345 350
Tyr Tyr Gly Phe Cys Asn Asp Lys Phe Lys Lys Lys Ala Gln Glu Arg
355 360 365
Gly Arg Leu Leu Thr Lys Ile Lys Ala Ile Arg Lys Glu Lys Ile Glu
370 375 380
Ser Gln Asn Leu Arg Tyr Trp Ser Leu Ile Leu Asp Asp Gly Lys Asp
385 390 395 400
Lys Phe Leu Trp Leu Val Pro Lys Glu Asn Met Gln Glu Phe Arg Arg
405 410 415
Glu Leu Ser Lys Ile His Pro Ser Gly Glu Ser Ser Leu Phe Leu Phe
420 425 430
His Ser Leu Thr Met Arg Ala Leu His Lys Leu Cys Phe Ala Gln Glu
435 440 445
Ser Asp Phe Val Lys Glu Met Pro Lys Val Leu Lys Glu Glu Gln Leu
450 455 460
Asn Cys Glu Lys Ala Ser Asn Asp Thr Glu Thr Asn Lys Arg Ile Lys
465 470 475 480
Arg Asn Phe Gly Leu Asn Tyr Ile Lys Thr Lys Asp Glu Leu Thr Leu
485 490 495
Ser Phe Leu Lys Lys Leu Ile Ile Ser Glu Tyr Ala His Glu Arg Leu
500 505 510
Asp Leu Asn His Phe Asp Leu Ser Lys Leu Gln Val Ala Thr Thr Leu
515 520 525
Asn Glu Phe Glu Glu Tyr Leu Glu Asp Ala Cys Tyr Tyr Leu Glu Lys
530 535 540
Ile Ser Ile Ser Ser Ser Met Ile Lys Glu Leu Leu Glu Glu Tyr Asn
545 550 555 560
Ile Leu Asn Phe Arg Ile Thr Ser Tyr Asp Leu Glu Lys Arg Asn Lys
565 570 575
Asn Thr Tyr Gln Thr Pro Glu Ser Asp Ile Lys Arg His Thr Lys Glu
580 585 590
Ile Trp Asn Lys Phe Trp Glu Gly Asp Arg Phe Ile Arg Leu Asn Pro
595 600 605
Glu Ile Lys Ile Arg Tyr Arg Gln Lys Asn Gln Asn Ile Glu Asp Tyr
610 615 620
Leu Lys Glu Lys Gly Phe Asp Leu Thr Lys Ile Lys Asn Arg Phe Leu
625 630 635 640
Gln Glu Gln Tyr Ser Val Ser Phe Thr Phe Ala Leu Asn Ala Gly Lys
645 650 655
Lys Tyr Pro Lys Leu Ala Phe Val Lys Thr Glu Glu Ile Leu Glu Lys
660 665 670
Ile Glu Glu Phe Asn Asp Glu Phe Asn Lys Gln Tyr Phe Asp Asn Ser
675 680 685
Tyr Lys Tyr Gly Ile Asp Arg Gly Asn Ile Glu Leu Ala Thr Leu Cys
690 695 700
Ile Thr Lys Phe Asn Lys Asn Asp Thr Tyr Glu Tyr Lys Gly Lys Lys
705 710 715 720
Tyr Leu Lys Pro Asn Phe Pro Thr Ser Gln Glu Asp Ile Lys Thr Tyr
725 730 735
Glu Leu Lys Asn Glu Trp Tyr Lys Arg Thr Ala Ile Ser Asn Ile Glu
740 745 750
Thr Lys Pro Lys Asn Lys Lys Thr Pro Lys Arg Ile Ile Ala Asn Ile
755 760 765
Ser Tyr Phe Ile Asp Asn Val Glu Asn Glu Glu Trp Phe Asn Lys Lys
770 775 780
Thr Cys Thr Ser Ile Asp Leu Thr Thr Ala Lys Val Ile Lys Gly Lys
785 790 795 800
Leu Ile Leu Asn Gly Asp Val Leu Thr Phe Leu Lys Leu Lys Lys Glu
805 810 815
Ala Ala Lys Arg Ile Leu Phe Glu Leu Val Ala Gln Asn Lys Leu Thr
820 825 830
Ala Lys Asn Lys Glu Leu Lys Trp Lys Ser Asp Asp Gly Asn Asn Ser
835 840 845
Asp Ser Val Arg Leu Ile Cys Asp Val Leu Asp Asn Glu Thr Asn Ser
850 855 860
Ile Tyr Phe Tyr Glu Asp Ser Lys Tyr Gly Arg Gly Phe Glu Gly Leu
865 870 875 880
Leu Thr Thr Asp Lys Thr Ala Tyr Ser Lys Glu Gly Ile Arg Ile Asn
885 890 895
Leu Gln Asn Tyr Leu Asn His Leu Ile Ser Glu Lys Glu Asn Lys Ser
900 905 910
Asn Lys Ala Tyr Ser His Val Pro Ser Ile Glu Lys Ile Asn His Leu
915 920 925
Arg Asp Ala Leu Val Ala Asn Met Val Gly Val Ile Ser Tyr Leu Gln
930 935 940
Ala Tyr Tyr Pro Gly Ile Val Val Leu Glu Asp Leu Asn His Lys Leu
945 950 955 960
Leu Ile Lys His Phe Glu Asp Leu Asn Ile Asn Ile Ser Asn Arg Phe
965 970 975
Glu His Ala Leu Ile Glu Lys Phe Gln Thr Leu Gly Met Val Pro Pro
980 985 990
His Ile Lys Asp Tyr Leu Glu Ile Arg Ser Ser Phe Arg Met Ser Arg
995 1000 1005
Asn Asp Ser Ser Gln Phe Gly Ala Leu Ile Phe Val Ser Lys Glu
1010 1015 1020
Gly Thr Ser Lys Glu Cys Pro Tyr Cys Glu Lys Lys Trp Asn Trp
1025 1030 1035
Gly Lys Glu Lys Glu Ile Glu Leu Lys Phe Ser Lys Lys Gln Tyr
1040 1045 1050
Ile Cys Gly Lys Glu Asn Ser Cys Gly Phe Asp Thr Lys His Ile
1055 1060 1065
Gln Asn Thr Phe Glu Phe Leu Ser Glu Ile Asn Asp Pro Asp Lys
1070 1075 1080
Ile Ala Ala Tyr Asn Ile Ala Lys Arg Gly Phe Lys Ser Phe Ile
1085 1090 1095
Asn Lys Ser Ser Ile Lys Lys Gln Ser Arg Ala Asp Pro Lys Lys
1100 1105 1110
Lys Arg Lys Val
1115
<210> 87
<211> 1096
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.8-NLS融合蛋白的氨基酸序列
<400> 87
Met Glu Lys Tyr Lys Ile Thr Lys Thr Ile Arg Phe Lys Leu Leu Pro
1 5 10 15
Asp Lys Ile Gln Asp Ile Ser Arg Gln Val Ala Val Leu Gln Asn Ser
20 25 30
Thr Asn Ala Glu Lys Lys Asn Asn Leu Leu Arg Leu Ile Gln Arg Gly
35 40 45
Gln Glu Leu Pro Lys Leu Leu Asn Glu Tyr Ile Arg Tyr Ser Asp Asn
50 55 60
His Lys Leu Lys Ser Asn Val Thr Val His Phe Arg Trp Leu Arg Leu
65 70 75 80
Phe Thr Lys Asp Leu Phe Tyr Asn Trp Lys Lys Asp Asn Thr Glu Lys
85 90 95
Lys Ile Lys Ile Ser Asp Val Asp Tyr Leu Ser Arg Val Phe Glu Asp
100 105 110
Phe Phe Asn Glu Trp Glu Thr Val Ile Glu Arg Ile Asn Thr Asp Cys
115 120 125
Asn Arg Pro Glu Glu Ser Lys Thr Arg Asp Ala Glu Ile Ala Phe Ser
130 135 140
Ile Lys Lys Ile Ala Thr Lys Gln Met Phe Pro Phe Ile Lys Ser Phe
145 150 155 160
Val Tyr Asn Ser Asn Tyr Lys Asn Ser Glu Glu Thr Lys Ser Lys Leu
165 170 175
Thr Ala Leu Leu Asn Glu Phe Glu Thr Val Leu Lys Ile Cys Glu Gln
180 185 190
Asn Tyr Leu Pro Ser Gln Ser Ala Gly Ile Val Ile Ala Lys Ala Ser
195 200 205
Phe Asn Tyr Tyr Thr Ile Asn Lys Lys Gln Lys Asp Tyr Lys Gly Tyr
210 215 220
Thr Asp Asp Ile Glu Lys Ile Glu Lys Gly Met Asn Ser Lys Phe His
225 230 235 240
Tyr Glu Arg Lys Tyr Asp Gln Leu Leu Glu Glu Leu Asn Leu Ile Ala
245 250 255
Leu Lys Glu Leu Pro Leu Ile Glu Phe Tyr Ser Lys Ile Lys Ser Tyr
260 265 270
Lys Ser Thr Arg Lys Ile Glu Phe Ser Glu Ala Val Ser Lys Gly Leu
275 280 285
Ala Phe Ala Asp Leu Lys Ser Lys Phe Pro Leu Phe Gln Thr Glu Ser
290 295 300
Asn Lys Tyr Ala Glu Phe Leu Glu Leu Thr Gly Arg Ile Thr Gln Ile
305 310 315 320
Ser Thr Ala Lys Ser Leu Leu Ser Lys Asp Asn Pro Glu Ala Gln Lys
325 330 335
Leu Arg Asp Glu Ile Lys Lys Leu Arg Ile Asn Arg Gly Glu Tyr Phe
340 345 350
Lys Asn Asn Phe His Lys Tyr Ile Ser Leu Cys Asn Leu Tyr Lys Lys
355 360 365
Ile Ala Asp Lys Lys Gly Arg Leu Lys Gly Gln Val Lys Gly Ile Glu
370 375 380
Asn Glu Arg Ile Asp Ser Gln Arg Ile Gln His Trp Ala Leu Val Leu
385 390 395 400
Glu Asp Asn Leu Lys His Ser Leu Ile Leu Ile Pro Lys Glu Lys Val
405 410 415
Thr Glu Val Tyr Arg Lys Val Arg Ala Ser Lys Ala Asp Ser Thr Ser
420 425 430
Ser Ser Ser Ser Leu Tyr Tyr Phe Glu Ser Met Thr Tyr Arg Ala Leu
435 440 445
His Lys Leu Cys Phe Gly Val Asn Gly Asn Thr Phe Leu Pro Glu Ile
450 455 460
Gln Lys Glu Leu Pro Glu Tyr Asn Pro Asn Lys Gln Ser Asp Phe Gly
465 470 475 480
Glu Phe Cys Phe His Lys Ser Asn Thr Asp Lys Glu Ile Asp Glu Pro
485 490 495
Lys Leu Ile Ser Phe Tyr Gln Ser Val Leu Lys Thr Asn Tyr Val Lys
500 505 510
Asp Asn Leu Asn Leu Pro Gln Ser Val Phe Asp Glu Ala Thr Val Gln
515 520 525
Thr Phe Glu Thr Arg Gln Asp Phe Gln Ile Ala Leu Glu Lys Cys Cys
530 535 540
Tyr Ala Lys Lys Thr Ile Ile Ser Glu Thr Leu Lys Lys Glu Ile Leu
545 550 555 560
Glu Asp Asn Asn Val Gln Ile Phe Gln Ile Thr Ser Leu Asp Leu Gln
565 570 575
Arg Ser Glu Gln Lys Asn Leu Lys Ala His Thr Lys Ile Trp Asn Arg
580 585 590
Phe Trp Thr Lys Gln Asn Glu Thr Ala Asn Tyr Asp Leu Arg Leu Asn
595 600 605
Pro Glu Thr Ala Ile Val Trp Arg Lys Pro Lys Lys Thr Arg Ile Asp
610 615 620
Lys Tyr Gly Ala Gly Thr Ser Leu Tyr Asp Pro Lys Lys Arg Asn Arg
625 630 635 640
Tyr Leu His Glu Gln Tyr Thr Leu Cys Thr Thr Val Thr Asp Asn Ala
645 650 655
Leu Asn Asn Glu Ile Thr Phe Ala Phe Glu Asp Thr Lys Lys Lys Gly
660 665 670
Thr Glu Ile Val Lys Tyr Asn Glu Lys Ile Asn Gln Thr Leu Lys Lys
675 680 685
Glu Phe Asn Lys Asn Gln Leu Trp Phe Tyr Gly Ile Asp Ala Gly Glu
690 695 700
Ile Glu Leu Ala Thr Leu Ala Leu Met Asn Lys Asp Lys Glu Pro Gln
705 710 715 720
Leu Phe Thr Val Tyr Glu Leu Lys Lys Ser Asp Phe Phe Lys His Gly
725 730 735
Tyr Ile Tyr Asn Lys Glu Arg Glu Leu Val Ile Arg Glu Lys Pro Tyr
740 745 750
Lys Ala Ile Gln Asn Leu Ser Tyr Phe Leu Asn Glu Glu Leu Tyr Glu
755 760 765
Lys Thr Phe Arg Asp Gly Lys Phe Gln Glu Thr Phe Asn Glu Leu Phe
770 775 780
Lys Glu Lys His Val Ser Ala Ile Asp Leu Thr Thr Ala Lys Val Ile
785 790 795 800
Asn Gly Lys Ile Ile Leu Asn Gly Asp Met Ile Thr Phe Leu Asn Leu
805 810 815
Arg Ile Leu His Ala Lys Arg Lys Ile Tyr Glu Glu Leu Ile Ile Asn
820 825 830
Pro Gln Ala Glu Leu Lys Glu Asn Glu Lys Glu Tyr Tyr Leu Tyr Phe
835 840 845
Asp Lys Glu Gly Thr Glu Lys Val Glu Lys Ile Tyr Arg Ser Arg Leu
850 855 860
Asp Phe Glu His Ile Lys Pro Tyr Gln Glu Ile Arg Asn Asp Leu Asn
865 870 875 880
Ala Tyr Phe Lys Asn Val Gln Lys Asn Glu Ala Lys Val Glu Asp Gln
885 890 895
Ile Asn Gln Thr Arg Arg Ala Leu Val Gly Asn Met Ile Gly Val Ile
900 905 910
Tyr Tyr Leu Tyr Gln Lys Tyr Arg Gly Ile Ile Ser Ile Glu Asp Leu
915 920 925
Lys Gln Thr Lys Val Glu Ser Asp Arg Asn Lys Phe Glu Gly Asn Ile
930 935 940
Glu Arg Pro Leu Glu Trp Ala Leu Tyr Arg Lys Phe Gln Gln Glu Gly
945 950 955 960
Tyr Val Pro Pro Ile Ser Glu Leu Ile Lys Leu Arg Glu Leu Glu Lys
965 970 975
Phe Pro Leu Lys Asp Val Lys Gln Pro Lys Tyr Glu Asn Ile Gln Gln
980 985 990
Phe Gly Ile Ile Lys Phe Val Ser Pro Glu Glu Thr Ser Thr Thr Cys
995 1000 1005
Pro Ser Cys Glu Lys Lys Ala Tyr Glu Leu Gln Lys Glu Lys Lys
1010 1015 1020
Gly Glu Glu Lys Pro Ala Glu Asn Lys Arg Tyr Glu Ala Asp Lys
1025 1030 1035
Lys Ala Gly Val Phe Cys Cys Pro Lys Cys Gly Phe His Asn Arg
1040 1045 1050
Thr Asn Pro Met Gly Tyr Glu Ser Leu Asp Ser Asn Asp Lys Val
1055 1060 1065
Ala Ala Phe Asn Ile Ala Lys Arg Gly Phe Glu Asp Leu Gln Lys
1070 1075 1080
His Lys Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val
1085 1090 1095
<210> 88
<211> 1044
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.9-NLS融合蛋白的氨基酸序列
<400> 88
Met Glu Asn Ser Asn Leu Tyr Gln Val Val Lys Thr Ile Arg Phe Lys
1 5 10 15
Leu Glu Pro Val Gly Lys Met Asp Thr Pro Lys Phe Gly Asp Lys Asn
20 25 30
Ala Glu Ser Lys Ala Asn Leu Thr Pro Phe Ile Glu Leu Val Lys Lys
35 40 45
Thr Met Thr Asn Val Lys Ala Leu Val Phe Ser Lys Gln Asp Gly Glu
50 55 60
Asp Gly Glu Lys Trp Arg Lys Ile Leu Glu Val Asn Tyr Arg Phe Leu
65 70 75 80
Arg Ser Tyr Leu Lys Asn Ser Phe Tyr Glu Asn Arg Gly Asp Ser Gln
85 90 95
Glu Lys Ser Lys Lys His Lys Ile Ser Asp Leu Glu Tyr Leu Gln Lys
100 105 110
Ala Leu Glu Asn Leu Phe Ala Glu Phe Asp Glu Ile Leu Asp Gly Leu
115 120 125
Glu Asp Phe Glu Lys Arg Asn Thr Lys Asn Gln Tyr Glu Lys Gln Arg
130 135 140
His Ala Gln Ala Gly Leu Leu Leu Asn Arg Leu Cys Lys Arg Ser Asn
145 150 155 160
Phe Gly Phe Leu Lys Ala Phe Val Gly Ala Leu Ala Gln Thr Asn Lys
165 170 175
Pro Phe Phe Asp Asp Lys Thr Asp Lys Leu Lys Lys Gln Ile Asp Lys
180 185 190
Phe Glu Thr Glu Leu Glu Lys Gln Lys Glu Phe Phe Leu Pro Tyr Gln
195 200 205
Ser Asn Gly Val Leu Phe Ala Gly Gly Ser Phe Asn Arg Tyr Ala Ile
210 215 220
Asn Lys Thr Pro Lys Met Leu Asp Lys Glu Leu Arg Glu Glu Gln Thr
225 230 235 240
Asn Leu Lys Lys Ser Leu Cys Glu His Lys Ile Lys Ile Asp Thr Leu
245 250 255
Asn Thr Leu Gly Leu Lys Asn Asp Cys Pro Cys Thr Ser Leu Asp Asn
260 265 270
Ser Tyr Thr Phe Ile Lys Asp Tyr Lys Ala Lys Gln Lys Ser Lys Phe
275 280 285
Ile Glu Leu Val Gln Lys Gly Glu Phe Asp Glu Ala Lys Lys Val Asn
290 295 300
Leu Phe Glu Cys Ser Glu Thr Asp Phe Glu Thr Phe Lys Thr Arg Thr
305 310 315 320
Lys Gln Ile Gln Asn Glu Lys Asp Lys Asp Glu Arg Thr Lys Leu Lys
325 330 335
Gln Lys Arg Gly Glu Phe Phe Lys Ser Gln Lys Arg Gly Lys Phe Phe
340 345 350
Lys Ser Gln Thr Gln Asn Tyr Glu Asn Leu Cys Asp Leu Tyr Lys Lys
355 360 365
Ile Ala Gln Lys Arg Gly Gln Ile Val Ala Lys Ile Cys Ala Ile Lys
370 375 380
Lys Glu Lys Glu Met Cys Glu Gln Val Lys Tyr Trp Cys Val Ala Leu
385 390 395 400
Glu Lys Gly Gly Glu Phe Tyr Leu Tyr Met Phe Leu Arg Asp Glu Asn
405 410 415
Asp Asn Ile Lys Asn Ala Tyr Asp Phe Val Ser Lys Leu Gln Thr Gln
420 425 430
Lys Ser Gly Glu Thr Lys Leu His Tyr Phe Asp Ser Leu Thr Leu Lys
435 440 445
Ala Val Arg Lys Leu Cys Phe Lys Glu Thr Asp Gly Ser Phe Lys Lys
450 455 460
Ala Leu Lys Asn Val Lys Phe Pro Glu Cys Glu Gln Asn Leu Asp Glu
465 470 475 480
Lys Val Lys Ile Ser Phe Tyr Gln Asn Val Leu Lys Asn Ala Lys Thr
485 490 495
Leu Asn Leu Ser Lys Phe Glu Asn Leu Gln Ser Val Thr Glu Gly Lys
500 505 510
Phe Glu Ser Leu Ser Glu Phe Glu Val Ala Leu Asn Met Val Cys Tyr
515 520 525
Thr Lys Thr Val Cys Val Ser Glu Ser Val Glu Lys Glu Leu Lys Lys
530 535 540
Phe Lys Pro Leu Val Phe His Ile Thr Ser Gln Asp Leu Ala Ala Lys
545 550 555 560
Arg Glu Lys Lys Ala His Thr Gln Ile Trp His Glu Phe Trp Arg Glu
565 570 575
Ser Asn Glu Lys Ser Lys Phe Pro Leu Arg Leu Asn Pro Glu Leu Lys
580 585 590
Val Met Trp Arg Glu Ala Arg Pro Ser Arg Val Glu Lys Tyr Ala Glu
595 600 605
Gln Ser Asp Lys Phe Asp Pro Asn Lys Lys Asn Arg Tyr Leu His Pro
610 615 620
Gln Phe Thr Leu Ala Leu Asn Phe Thr Gln Asn Ala His Asn Glu Ala
625 630 635 640
Ile Asn Leu Ala Phe Lys Asp Val Gln Asn Lys Gly Glu Ala Val Lys
645 650 655
Lys Phe Asn Glu Asn Phe Lys Ser Ser Glu Tyr Ala Phe Gly Ile Asp
660 665 670
Val Gly Thr Lys Asp Leu Ala Leu Leu Cys Leu Ile Asp Lys Asn Lys
675 680 685
Lys Pro Val Asn Phe Asp Val Tyr Glu Ile Cys Asn Glu Asn Glu Ile
690 695 700
Cys Asn Glu Lys Leu Gly Phe Glu Lys Phe Gly Phe Tyr Lys Asp Gly
705 710 715 720
Thr Arg Arg Asp Glu Pro Tyr Lys Leu Ile Lys Asn Pro Ser Tyr Phe
725 730 735
Leu Asn Glu Ser Leu Tyr Lys Lys Thr Phe Asn Ala Thr Lys Glu Glu
740 745 750
Phe Glu Arg Ser Phe Ser Glu Leu Phe Lys Arg Lys Ser Val Cys Ala
755 760 765
Leu Asp Leu Thr Thr Ala Lys Val Ile Cys Gly Lys Ile Ile Leu Asn
770 775 780
Gly Asp Phe Ser Thr His Leu Asn Leu Lys Ile Leu Asn Ala Lys Arg
785 790 795 800
Lys Ile Ser Ala Lys Leu Lys Lys Asp Pro Thr Leu Lys Ile Glu Tyr
805 810 815
Asp Asn Asp Asp Asn Ile Leu Phe Gly Ser Asn Val Ile Phe Tyr Tyr
820 825 830
Asn Asn Lys Tyr Glu Ile Val Arg Pro Tyr Asp Glu Ile Lys Asn Glu
835 840 845
Ile Phe Glu Phe His Glu Lys Gln Arg Leu Asp Asp Ala Arg Leu Glu
850 855 860
Asp Asn Ile Asn Lys Thr Arg Ala Asn Leu Val Ala Asn Met Val Gly
865 870 875 880
Val Ile Ser Phe Leu His Lys Glu Phe Ser Gly Phe Val Val Leu Glu
885 890 895
Asn Leu Lys Gln Ser Glu Ile Glu Gly Asn His Arg Leu Lys Phe Glu
900 905 910
Gly Asp Ile Thr Arg Pro Leu Glu Leu Ala Leu Tyr Arg Lys Phe Gln
915 920 925
Ser Lys Cys Leu Thr Pro Pro Ile Ser Glu Leu Ile Lys Leu Arg Glu
930 935 940
Gly Glu Lys Asn Glu Asn Val Glu Ser Asp Leu Ile Leu Gln Phe Gly
945 950 955 960
Ile Ile Lys Phe Val Asp Lys Asp Lys Thr Ser Arg Leu Cys Pro Ala
965 970 975
Cys Gly Lys Asp Ala Tyr Glu Asn Asn Asn Ser Lys Tyr Lys Thr Asp
980 985 990
Lys Lys Asp Gly Val Phe Glu Cys Ala Gly Cys Gly Phe Asn Asn Lys
995 1000 1005
Asn Asn Ala Gly Asp Phe Ala Ala Leu Asp Thr Asn Asp Lys Ile
1010 1015 1020
Ala Thr Phe Asn Ile Ala Lys Arg Gly Leu Ser Arg Ala Asp Pro
1025 1030 1035
Lys Lys Lys Arg Lys Val
1040
<210> 89
<211> 1179
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.10-NLS融合蛋白的氨基酸序列
<400> 89
Met Glu Thr Tyr Lys Ile Thr Lys Thr Ile Arg Phe Lys Leu Glu Ala
1 5 10 15
Asp Glu Glu Asn Ser Ile His Ile Lys Glu Asp Ile Ile Asn Ile Glu
20 25 30
Thr Asn Asp Asn Glu Phe Thr Met Val Asp Phe Val Ser Asn Leu Gly
35 40 45
Asn Tyr Ile Lys Asp Leu Lys Asn Tyr Leu Phe Tyr Glu Lys Lys Asp
50 55 60
Gly Ser Leu Ser Phe Lys Asp Lys Ile Ile Ile Lys Asn Glu Trp Leu
65 70 75 80
Arg Gln Tyr Ala Lys Gln Asp Phe Val Glu Leu Lys Ser Lys Lys Arg
85 90 95
Ile Asn Leu Arg Asn Asn Arg Met Glu Gln Ile Lys Ile Gly Asp Ile
100 105 110
Pro Arg Leu Ser Ser Lys Ile Glu Glu Ala Leu Asp Ile Ala Lys Glu
115 120 125
Ile Tyr Ser Lys Leu Ser Asp Asp Ala Thr Leu Glu Gln His Glu Arg
130 135 140
Thr Lys Lys Ala Gln Ile Gly Leu Leu Leu Lys Arg Leu Glu Ala Lys
145 150 155 160
Asn Val Leu Pro Leu Leu Met Asp Leu Val Lys Glu Thr Leu Asp Lys
165 170 175
Asp Glu Thr Asp Asp Leu Ser Ile Arg Leu Lys Arg Gln Ser Gln Lys
180 185 190
Ile Asn Ser Gln Leu Lys Ile Ala Ile Arg Ser Phe Leu Pro Glu Gln
195 200 205
Ser Asn Gly Leu Gln Ile Ala Lys Ala Ser Phe Asn Tyr Tyr Thr Ile
210 215 220
Asn Lys Lys Pro Ile Asp Phe Glu Lys Lys Ile Glu Asp Leu Lys Lys
225 230 235 240
Asn Leu Asn Val Lys Asp Leu Glu Lys Leu Asn Val Tyr Phe Asp Lys
245 250 255
Lys Glu Lys Lys Gln Lys Asn Tyr Leu Gly Lys Lys Ile Phe Ser Leu
260 265 270
Phe Glu Thr Asp Ile Gln Lys Ala Leu Ser Lys Asn Gln Pro Leu Tyr
275 280 285
Leu Gly Asp Ala Pro Met Ile Asp Ser Ala Tyr Val Ser Leu Arg Gln
290 295 300
Ile Phe Lys Lys Ile Lys Ser Glu Gln Lys Lys Gln Phe Ser Glu Leu
305 310 315 320
Met Gln Asn Lys Cys Ser Tyr Asp Glu Leu Lys Asn Ser Asn Leu Tyr
325 330 335
Leu Leu Asn Asp Ile Gly Leu Glu Gln Phe Asn Thr Tyr Arg Glu Lys
340 345 350
Thr Lys Glu Leu Glu Glu Leu Ala Thr Lys Leu Ser Asn Gln Asn Leu
355 360 365
Leu Glu Asn Ala Lys Glu Arg Leu Arg Ser Gln Lys Glu Lys Ile Ala
370 375 380
Lys Glu Arg Gly Asn Ile Met Lys Asp Arg Phe Gln Thr Trp Lys Ser
385 390 395 400
Phe Ala Asn Phe Tyr Arg Thr Val Ser Gln Lys His Gly Lys Ile Leu
405 410 415
Ala Gln Leu Lys Gly Ile Glu Lys Glu Gln Ala Glu Ser Gln Leu Leu
420 425 430
Lys Tyr Trp Ala Leu Ile Cys Glu Lys Glu Asn Gln His Gln Leu Trp
435 440 445
Leu Ile Pro Arg Glu Lys Ala Trp Glu Cys Lys Arg Trp Leu Glu Thr
450 455 460
Val Asn Asp Thr Ser Ile Asp Asn Glu Asn Ser Ile Lys Leu Tyr Trp
465 470 475 480
Phe Glu Ser Leu Thr Tyr Arg Ser Leu Gln Lys Leu Cys Phe Gly Phe
485 490 495
Leu Glu Asn Gly Asn Asn Glu Phe Asn Gln Asn Ile Lys Asp Leu Leu
500 505 510
Pro Lys Asp Arg Ile Gly Asn Thr Ile Asn Gly Glu Phe Ala Phe Glu
515 520 525
Gly Asp Glu Glu Arg Lys Ile Glu Phe Tyr Lys Thr Val Leu Asn Ser
530 535 540
Lys Tyr Ala Lys Gln Val Leu Asn Ile Pro Phe Lys Gln Val Glu Glu
545 550 555 560
Glu Ile Ile Ser Gln Ser Phe Glu Asn Leu Ser Asp Phe Gln Ile Ala
565 570 575
Leu Glu Lys Ile Cys Tyr Arg Arg Phe Ala Ile Tyr Ser Asn Tyr Ile
580 585 590
Ile Ser Phe Asp Ala Gln Ile Phe Asp Ile Thr Ser Leu Asp Leu Lys
595 600 605
Asn Asn Glu Lys Asn Asn Leu Asn Thr His Thr His Ile Trp Arg Asp
610 615 620
Phe Trp Lys Asp Glu Asn Glu Lys Asn Asn Phe Asp Ile Arg Leu Asn
625 630 635 640
Pro Glu Ile Thr Ile Ser Tyr Arg Thr Pro Lys Gln Ser Arg Ile Glu
645 650 655
Lys Tyr Gly Glu Lys Thr Lys Glu Tyr Asp Pro Asn Lys Asn Asn Arg
660 665 670
Tyr Leu His Pro Gln Phe Thr Leu Ile Thr Thr Ile Ser Glu Arg Ser
675 680 685
Asn Ser Gln Thr Lys Thr Leu Ser Phe Ile Glu Asp Glu Asp Phe Lys
690 695 700
Lys Ser Ile Asn Glu Phe Asn Lys Lys Leu Lys Lys Asp Asn Ile Lys
705 710 715 720
Phe Ala Phe Gly Ile Asp Asn Gly Glu Val Glu Leu Ser Thr Leu Gly
725 730 735
Val Tyr Leu Pro Thr Phe Glu Lys Glu Thr His Glu Glu Lys Ile Tyr
740 745 750
Glu Leu Lys Gln Ile Lys Lys Tyr Gly Phe Glu Val Leu Thr Ile Thr
755 760 765
Asp Leu Lys Tyr Lys Glu Thr Asp Tyr Asn Gly Asn Val Arg Lys Ile
770 775 780
Ile Gln Asn Pro Ser Tyr Phe Leu Lys Lys Glu Asn Tyr Ile Arg Thr
785 790 795 800
Phe Ser Lys Ser Glu Gln Glu Tyr Glu Glu Met Phe Ala Lys Leu Phe
805 810 815
Lys Lys Glu His Val Leu Ser Leu Asp Leu Thr Thr Ala Lys Met Ile
820 825 830
Cys Gly His Ile Val Thr Asn Gly Asp Val Pro Ala Leu Phe Asn Leu
835 840 845
Trp Leu Lys His Ala Gln Arg Asn Val Phe Glu Met Asn Asp His Thr
850 855 860
Val Lys Glu Thr Ala Lys Thr Ile Arg Leu Arg Asn Asn Glu Glu Leu
865 870 875 880
Thr Asp Asn Glu Lys Glu Lys Phe Ala Glu Phe Ile Ser Asp Gly Lys
885 890 895
Lys Phe Ala Lys Leu Thr Lys Glu Gly Lys Lys Ser Arg Tyr Leu Lys
900 905 910
Trp Ile Phe Glu Asp Arg Lys Glu Asn Ser Phe Thr Glu Asp Glu Asn
915 920 925
Lys Lys Phe Asn Asp Cys Gln Lys Lys Lys Gly Lys Tyr Asn Ser His
930 935 940
Ile Ile Phe Ala Ser Arg Phe Glu Gly Asp Glu Leu Lys Ser Val Thr
945 950 955 960
Pro Ile Phe Asp Cys Arg His Val Phe Lys Lys Arg Lys Glu Phe Glu
965 970 975
Thr Ile Arg Pro Ile Lys Glu Ile Glu Asn Glu Ile Ser Arg Phe Asn
980 985 990
Thr Asn Arg Thr Ser His Asn Ile Ser Asn Glu Glu Leu Asp Leu Lys
995 1000 1005
Ile Thr Asp Ala Lys Lys Ala Leu Val Ala Asn Ala Ile Gly Val
1010 1015 1020
Ile Asp Phe Leu Tyr Lys Gln Tyr Lys Gln Arg Phe Asn Asp Glu
1025 1030 1035
Gly Leu Ile Ile Lys Glu Gly Phe Asp Thr Gln Lys Val Glu Glu
1040 1045 1050
Asp Ile Glu Lys Phe Ser Gly Asn Ile Tyr Arg Ile Leu Glu Arg
1055 1060 1065
Lys Leu Tyr Gln Lys Phe Gln Asn Tyr Gly Leu Val Pro Pro Ile
1070 1075 1080
Lys Asn Leu Met Ala Val Arg Asn Glu Gly Ile Lys Asp Lys Asn
1085 1090 1095
Ala Ile Leu Arg Leu Gly Asn Ile Ala Phe Ile Asp Pro Ser Gly
1100 1105 1110
Thr Ser Gln Glu Cys Pro Val Cys Lys Glu Lys Ser Lys Glu Lys
1115 1120 1125
His Thr Asn Asn Phe Ile Cys Glu Cys Gly Phe Asn Ser Thr Asn
1130 1135 1140
Ile Met His Ser Asn Asp Gly Ile Ala Gly Phe Asn Ile Ala Lys
1145 1150 1155
Arg Gly Phe Glu Asn Phe Ile Asn Glu Lys Ser Arg Ala Asp Pro
1160 1165 1170
Lys Lys Lys Arg Lys Val
1175
<210> 90
<211> 1198
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.11-NLS融合蛋白的氨基酸序列
<400> 90
Met Glu Lys Tyr Lys Ile Thr Lys Thr Ile Arg Phe Arg Leu Asp Ala
1 5 10 15
Asp Asn Thr Ala Ile Ser Ala Ile Val Lys Asp Thr Glu Ala Leu Glu
20 25 30
Ala Arg Gly Gln Gly Phe Lys Ile Lys Lys Phe Val Asn Ala Leu Gly
35 40 45
Arg Phe Leu Ser Gly Asp Gly Val Gln Lys Tyr Leu Tyr Asp Met Ser
50 55 60
Asn Glu Glu Asn Cys Val Phe Lys Arg Asn Leu Val Ile Lys Asn Thr
65 70 75 80
Trp Leu Lys Asn Asn Ala Lys Gln Glu Ile Ala Gly Met Asp Leu Lys
85 90 95
Arg Gly Leu Ile Ile Lys Asp Ile Lys Gly Leu Gln Asp Lys Ile Glu
100 105 110
Glu Ile Tyr Asp Lys Leu Trp Glu Ile Tyr Glu Ile Leu Tyr Glu Ser
115 120 125
Ala Tyr Leu Pro Leu Gln Asp Leu Ala Arg Arg Glu Gly Ile Gly Leu
130 135 140
Leu Leu Lys Lys Leu Ser Val Lys Asn Ala Leu Pro Phe Ile Ile Ser
145 150 155 160
Phe Val Glu Glu Ser Asn Asp Lys Asn Glu Ala Asp Asp Leu Ser Leu
165 170 175
Arg Leu Lys Lys Gln Gly Lys Glu Ile Leu Thr Gln Leu Glu Ile Gly
180 185 190
Ile Asn Glu Tyr Leu Pro Ala Gln Ser Ser Gly Leu Pro Val Ala Lys
195 200 205
Ala Ser Phe Asn Tyr Tyr Thr Ile Asn Lys Thr Pro Val Asp Phe Gly
210 215 220
Glu Lys Ile Gln Glu Leu Glu Lys Arg Leu Ser Val Asp Ile Lys Lys
225 230 235 240
Glu Ile Ser Ser Phe Thr Gly Gly Ile Lys Thr Ala Ile Lys Asn Lys
245 250 255
Ile Ala Gly Lys Lys Ile Leu Leu Gly Asp Thr Pro Met Phe Glu Ser
260 265 270
Glu Asn Ser Val Ser Leu Arg Gln Ile Leu Lys Asn Ile Lys Ser Glu
275 280 285
Gln Lys Ala Gln Phe Asn Lys Phe Met Thr Thr Gln Asn Asn Pro Gln
290 295 300
Leu Glu Glu Met Lys Thr Met Gly Trp Tyr Leu Phe Gly Asp Ile Thr
305 310 315 320
Glu Gly Glu Phe Asn Asp Tyr Lys Glu Gln Thr Lys Glu Ile Glu Arg
325 330 335
Val Gly Ala Lys Ile Asn Gln Cys Gly Asn Ile Lys Glu Lys Lys Glu
340 345 350
Leu Arg Ser Gln Leu Gln Lys Leu Lys Lys Lys Arg Gly Glu Leu Ile
355 360 365
Ser Glu Ala His Lys Lys Gly Gly Asn Asp Lys Asn Phe Lys Thr Tyr
370 375 380
Lys Glu Phe Ala Lys Phe Tyr Arg Lys Ile Ala Gln Arg His Gly Lys
385 390 395 400
Ile Leu Ala Gln Ile Lys Gly Ile Glu Lys Glu Lys Ile Asp Ser Ala
405 410 415
Met Leu Asn Tyr Trp Ala Ala Val Ile Glu Leu Ser Gly Arg His Lys
420 425 430
Leu Val Leu Ile Pro Lys Lys Asp Glu Asn Ala Lys Lys Cys Ile Glu
435 440 445
Trp Leu Glu Asp Glu Ser Lys His Lys Asn Gly Ser Cys Lys Ile Phe
450 455 460
Trp Phe Glu Ser Phe Thr Phe Arg Ser Leu Gln Lys Leu Cys Phe Gly
465 470 475 480
Asn Leu Asp Ser Gly Thr Asn Thr Phe Asn Gln Lys Ile Gln Asn Leu
485 490 495
Leu Pro Cys Asp Glu Arg Gly Asn Leu Met Asn Gly Glu Phe Ala Phe
500 505 510
Lys Gly Asp Glu Gln Glu Lys Ile Lys Phe Tyr Lys Lys Val Leu Gln
515 520 525
Ser Gln Lys Asp Ile Asn Leu Pro Gln Lys Glu Val Val Asp Asn Val
530 535 540
Val Gly Arg Lys Phe Glu Thr Met Asp Glu Phe Lys Ile Ala Leu Glu
545 550 555 560
Glu Ile Cys Tyr Ile Arg Arg Glu Arg Leu Ser Ala Asn Ala Glu Ser
565 570 575
Glu Leu Lys Ser Lys Phe Asn Ala Gln Ile Phe Asp Ile Thr Ser Leu
580 585 590
Asp Leu Arg Asn Pro Val Asn Cys Ala Gly Lys Pro Glu Val Tyr His
595 600 605
His Asn Asp Lys Arg His Thr Glu Ile Trp Lys Glu Phe Trp Ser Leu
610 615 620
Asp Asn Glu Arg Arg Asn Phe Asn Ile Arg Leu Asn Pro Glu Ile Thr
625 630 635 640
Ile Thr Tyr Arg Lys Pro Lys Glu Ser Arg Ile Leu Lys Tyr Gly Lys
645 650 655
Gly Thr Glu Lys Tyr Asn Ala Asp Met Lys Asn Arg Tyr Leu Tyr Pro
660 665 670
Gln Tyr Thr Leu Leu Thr Thr Ile Ser Glu His Cys Asn Thr Pro Thr
675 680 685
Lys Ile Leu Ser Phe Met Thr Asp Asn Glu Tyr Glu Glu Ser Ile Lys
690 695 700
Ala Phe Asn Ser Lys Leu Lys Lys Glu Asp Ile Lys Phe Ala Phe Gly
705 710 715 720
Ile Asp Ser Gly Glu Thr Glu Leu Ser Thr Leu Gly Val Tyr Leu Pro
725 730 735
Glu Phe Ser Ala Glu Ser Thr Glu Leu Lys Asp Ile Glu Lys Tyr Gly
740 745 750
Phe Asn Val Leu Thr Ile Lys Asp Leu Asn Tyr Thr Glu Thr Asp Tyr
755 760 765
Asn Gly Ser Asp Lys Lys Ile Val Lys Asn Pro Ser Tyr Phe Val Asp
770 775 780
Lys Ser Leu Tyr Met Arg Thr Phe Lys Lys Thr Glu Gln Glu Tyr Glu
785 790 795 800
Lys Met Phe Ala Glu Gln Phe Glu Ala Lys Lys Arg Leu Ser Leu Asp
805 810 815
Leu Ser Ala Ala Lys Val Ile Cys Gly His Ile Val Thr Asn Gly Gly
820 825 830
Val Ser Glu His Phe Gly Leu Trp Leu Lys His Ala Gln Arg Thr Ile
835 840 845
Phe Trp Met Asn Asp His Thr Glu Lys Lys Thr Ala Lys Asn Ile Lys
850 855 860
Leu Lys Asp Ser Ser Glu Leu Thr Tyr Asp Glu Arg Glu Lys Phe Ala
865 870 875 880
Glu His Ile Ser Ser Asp Glu Lys Phe Lys Lys Leu Asp Val Glu Glu
885 890 895
Lys Lys Arg Tyr Val Arg Trp Ile Phe Glu Asp Arg Glu Thr Leu Asn
900 905 910
Phe Thr Glu Ala Glu Asn Lys Lys Phe Gly Gly Tyr Gln Lys Lys Lys
915 920 925
Gly Asp Tyr Arg Leu Gly Ile Leu Phe Ala Ser Cys Phe Ile Gly Lys
930 935 940
Glu Leu Glu Ser Val Thr Gln Ile Leu Asp Cys Arg His Ile Phe Lys
945 950 955 960
Lys Arg Glu Glu Phe Tyr Ser Leu Lys Ser Lys Glu Asp Ile Glu Ala
965 970 975
Glu Ile Lys Arg Tyr Asn Thr Asp Tyr Thr Asn His Asn Ile Ser Thr
980 985 990
Glu Gln Leu Asp Leu Lys Phe Val Asn Val Lys Asn Ala Leu Val Ala
995 1000 1005
Asn Ala Val Gly Val Ile Asp Leu Leu Tyr Lys Gln Tyr Lys Glu
1010 1015 1020
Arg Leu Gly Gly Glu Gly Leu Ile Ala Lys Glu Gly Phe Asp Thr
1025 1030 1035
Lys Lys Val Glu Glu Asp Met Glu Lys Phe Ser Gly Asn Ile Tyr
1040 1045 1050
Arg Ile Leu Glu Arg Lys Leu Tyr Gln Lys Phe Gln Asn Tyr Gly
1055 1060 1065
Leu Val Pro Pro Ile Lys Asn Leu Met Ala Val Arg Ala Asp Lys
1070 1075 1080
Val Glu Ile Ser Glu Ala Glu Lys Ser Lys Ile Arg Glu Asn Cys
1085 1090 1095
Lys Ile Ser Lys Ile Asp Pro Glu Asn Glu Ile Ile Lys Arg Asn
1100 1105 1110
Lys Thr Leu Ile Leu Arg Leu Gly Ser Ile Ala Phe Val Asn Asp
1115 1120 1125
Ala Asp Thr Ser Gln Glu Cys Pro Ala Cys Gly Thr Lys Ser Lys
1130 1135 1140
Glu Lys His Val Asp Asn Phe Ile Cys Gly Cys Gly Phe Asn Ser
1145 1150 1155
Thr Gly Ile Ile His Ser Asn Asp Gly Val Ala Gly Phe Asn Ile
1160 1165 1170
Ala Lys Arg Gly Phe Val Asn Leu Met Glu His Glu Leu Arg Ser
1175 1180 1185
Arg Ala Asp Pro Lys Lys Lys Arg Lys Val
1190 1195
<210> 91
<211> 1206
<212> PRT
<213> 人工序列
<220>
<223> Cas12j.12-NLS融合蛋白的氨基酸序列
<400> 91
Met Glu Lys Tyr Lys Leu Thr Lys Thr Ile Arg Phe Lys Leu Lys Pro
1 5 10 15
Lys Asp Ile Ser Ala Ile Lys Arg Asp Val Glu Ala Leu Glu Gln Gln
20 25 30
Lys Phe Asp Leu Val Leu Phe Val Tyr Asn Leu His Asn Phe Ile Gly
35 40 45
Lys Leu Lys Glu Tyr Leu Phe Phe Gln Lys Glu Lys Asp Glu Phe Val
50 55 60
Ile Lys Asp Lys Leu Thr Ile Lys Lys Thr Trp Leu Lys Gln Tyr Ala
65 70 75 80
Lys Gln Glu Ile Ala Gly Leu Glu Leu Asn Arg Glu Gln Thr Leu Gly
85 90 95
Asn Ile Lys Gly Val Ser Ala Arg Ile Glu Arg Ala Val Asp Asp Val
100 105 110
Asn Lys Ile Tyr Val Glu Leu Ala Met Glu Ala Lys Leu Asn Glu Arg
115 120 125
Ala Lys Lys Ala Lys Thr Glu Gln Leu Ile Lys Arg Leu Asp Thr Arg
130 135 140
Asn Ala Leu Pro Leu Leu Val Ser Leu Ile Glu Gln Ser Ser Asp Lys
145 150 155 160
Tyr Glu Thr Gly Asn Leu Ser Ile Gln Leu Lys Arg Leu Gly Lys Arg
165 170 175
Leu Gln Thr Gln Leu Leu Ser Gly Ile Lys Lys Tyr Leu Ala Glu Gln
180 185 190
Ser Asn Gly Leu Pro Ile Ala Lys Ala Ser Phe Asn Tyr Tyr Ala Ile
195 200 205
Asn Lys Lys Pro Val Asp Tyr Ile Asp Lys Ile Lys Gln Leu Gln Lys
210 215 220
Asp Leu Glu Ile Lys Lys Asn Arg Arg Ser Glu Glu Arg Tyr Asp Lys
225 230 235 240
Lys Lys Arg Lys Asn Ile Lys Ile Phe Asn Asp Ser Lys Leu Trp Ile
245 250 255
Lys Ile Lys Lys Asp Ile Glu Lys Glu Arg Gly Asn Lys Thr Leu Ile
260 265 270
Leu Gly Tyr Ala Pro Met Ile Glu Pro Gly Asn Tyr Val Tyr Leu Arg
275 280 285
Gln Ile Leu Lys Asn Ile Lys Leu Glu Gln Lys Asn Lys Phe Ser Lys
290 295 300
Leu Met Gln Ser Lys Ser Leu Thr Phe His Asp Leu Asn Asn Asn Asn
305 310 315 320
Gln Leu Tyr Leu Phe Lys Asp Ile Leu Glu Gly Glu Phe Asn Lys Tyr
325 330 335
Lys Gln Lys Thr Asn Glu Ile Glu Thr Lys Ala Glu Lys Arg Asn Gln
340 345 350
Cys Asn Asn Asp Glu Leu Lys Arg Lys Leu Asn Ser Glu Leu Gln Gln
355 360 365
Leu Arg Lys Asp Arg Gly Ser Leu Ile Asn Ala Ala Asp Gly Arg Pro
370 375 380
Lys Gly Arg Phe Lys Thr Tyr Lys Tyr Phe Ala Asn Phe Tyr Arg Asn
385 390 395 400
Val Ala Gln Lys His Gly Arg Ile Leu Ser Thr Leu Lys Gly Ile Glu
405 410 415
Lys Glu Met Val Glu Ser Gln Leu Leu Lys Tyr Trp Thr Ile Ile Thr
420 425 430
Glu Glu Asn Asn Gln His Ser Leu Val Leu Ile Pro Lys Glu Arg Ala
435 440 445
Gly Glu Tyr Lys Lys Asp Leu Glu Asn Ser Ile Pro Ser Asp Pro Ser
450 455 460
Ser Lys Ile Lys Val Tyr Trp Phe Glu Ser Phe Thr Leu Arg Ser Leu
465 470 475 480
Arg Lys Leu Cys Phe Gly Tyr Val Asn Asn Asn Thr Gly Ser Asn Thr
485 490 495
Phe Tyr Pro Glu Leu Lys Lys Ser Asp Glu Leu Arg Lys Tyr His Asp
500 505 510
Glu Arg Gly Asn Phe Ile Lys Gly Glu Phe Tyr Phe Lys Gly Asp Glu
515 520 525
Gln Lys Ile Ile Gln Phe Tyr Lys Asp Val Leu Arg Ser Asn Tyr Ala
530 535 540
Gln Lys Val Leu Lys Phe Pro Lys Gln Gln Val Lys Asp Glu Leu Ile
545 550 555 560
Gly Arg Glu Phe Ser Ser Leu Asp Glu Phe Gln Ile Ala Leu Glu Lys
565 570 575
Ile Cys Tyr Gln Arg His Val Val Cys Ser Gln Lys Val Val Asp Ala
580 585 590
Leu Ser Arg Tyr Asn Ala Gln Ile Phe Leu Ile Thr Ser Leu Asp Leu
595 600 605
Gly Asn Pro Ala Asn Cys Val Asp Lys Pro Lys Gln Phe Ser His Phe
610 615 620
Asp Lys Lys His Thr Arg Ile Trp Lys Glu Phe Trp Ser Ser Lys Asn
625 630 635 640
Glu Thr Ala Asn Phe Asp Ile Arg Leu Asn Pro Glu Ile Val Ile Thr
645 650 655
Tyr Arg Gln Pro Lys Gln Ser Arg Ile Lys Lys Tyr Gly Pro Glu Ser
660 665 670
Thr Arg Tyr Asp Asp Arg Lys His Asn Arg Tyr Leu Tyr Pro Gln Phe
675 680 685
Thr Leu Ile Thr Thr Ile Ser Glu Tyr Ser Asn Ala Pro Thr Lys Ala
690 695 700
Leu Ser Phe Leu Thr Asp Glu Glu Phe Lys Gly Ala Val Asp Glu Phe
705 710 715 720
Asn Lys Lys Phe Lys Lys Glu Asn Ile Arg Phe Ser Leu Gly Ile Asp
725 730 735
Asn Gly Glu Thr Glu Leu Ser Thr Leu Gly Val Tyr Leu Pro Val Phe
740 745 750
Lys Lys Asp Ser Asn Glu Lys Val Val Ala Glu Leu Lys Lys Val Asn
755 760 765
Lys Tyr Gly Phe Asn Phe Leu Thr Ile Lys Asp Leu Ser His Val Glu
770 775 780
Lys Asp Lys Asn Gly Arg Val Arg Lys Ile Ile Gln Asn Pro Ser Tyr
785 790 795 800
Phe Leu Ser Lys Glu Gln Tyr Met Arg Thr Phe Gly Arg Thr Glu Gln
805 810 815
Glu Tyr Asn Asn Met Phe Ala Glu Gln Phe Glu Glu Lys Ala Phe Leu
820 825 830
Ser Leu Asp Leu Thr Thr Ala Lys Val Ile Asn Gly His Ile Val Thr
835 840 845
Asn Gly Asp Val Pro Thr Phe Leu Asn Leu Trp Met Arg His Ala Gln
850 855 860
Arg Asp Ile Trp Asp Met Asn Asp His Thr Lys Glu Lys Thr Ala Lys
865 870 875 880
Lys Ile Val Ile Lys Asn Asn Asp Glu Leu Thr Asp Ala Glu Lys Val
885 890 895
Lys Phe Val Glu Tyr Ile Ser Asp Glu Thr Asn Tyr Ala Lys Leu Asn
900 905 910
Phe Asn Glu Lys Lys Arg Tyr Val Leu Trp Ile Phe Glu Asn Arg Lys
915 920 925
Asn Ile Asn Phe Thr Asp Ala Glu Lys Lys Lys Phe Glu Pro Cys Gln
930 935 940
Lys Arg Lys Gly Asn Phe Ser Lys Asp Ile Leu Phe Ala Val Cys Tyr
945 950 955 960
Ile Gly Ser Glu Ile His Ser Val Thr Asn Ile Phe Asp Val Arg Asn
965 970 975
Ile Phe Lys Met Arg Lys Asp Phe Tyr Val Leu Lys Ser Glu Met Glu
980 985 990
Ile Lys Lys Glu Ile Glu Ser Tyr Asn Thr Thr Ala Gly Ile Gln Glu
995 1000 1005
Ile Ser Asn Glu Glu Leu Asp Leu Lys Ile Asn Arg Leu Lys Gln
1010 1015 1020
Ala Val Val Ala Asn Ala Val Gly Val Ile Asp Tyr Leu Tyr Ile
1025 1030 1035
Tyr Tyr Lys Lys Lys Thr Gly Gly Glu Gly Leu Ile Ile Lys Glu
1040 1045 1050
Gly Phe Asp Thr Lys Lys Val Ala Lys Ala Leu Glu Lys Phe Ser
1055 1060 1065
Gly Asn Ile Tyr Arg Ile Leu Glu Arg Lys Leu Tyr Gln Lys Phe
1070 1075 1080
Gln Asn Tyr Gly Leu Val Pro Pro Ile Lys Ser Leu Met Ala Val
1085 1090 1095
Arg Glu Glu Gly Ile Glu Asn Asn Lys Asp Ala Ile Leu Arg Leu
1100 1105 1110
Gly Asn Val Gly Phe Ile Asp Pro Thr Gly Thr Ser Gln Gln Cys
1115 1120 1125
Pro Val Cys Ser Lys Gly Lys Leu Asn His Thr Thr Lys Cys Ser
1130 1135 1140
Lys Asn Cys Gly Phe Asn Ser Lys Asn Ile Met His Ser Asn Asp
1145 1150 1155
Gly Ile Ala Gly Tyr Asn Ile Ala Lys Arg Gly Phe Glu Asn Phe
1160 1165 1170
Ile Ser Gln Lys Lys Gly Tyr Asp Val Ile Asn Asn Gly Thr Lys
1175 1180 1185
Tyr Asn Asn Leu Lys Ser Gln Ser Arg Ala Asp Pro Lys Lys Lys
1190 1195 1200
Arg Lys Val
1205
<210> 92
<211> 3069
<212> DNA
<213> 人工序列
<220>
<223> Cas12g.1***表达盒的核苷酸序列
<400> 92
tttacacttt atgcttccgg ctcgtatgtt aggaggtctt tatcatgtta tataccatga 60
acgtaaaaac tataaaacta aaggtagacg ctacaaaaga agtagaatca agactgacaa 120
aaatgttgtt agtacacaat aacattggtc gtgaaattat taattttttg attttatgta 180
gtggaaatga taatatcaga aaaaccaaat ttgatgagtt tggaaattca tacgatgaat 240
tttgtaattt aaaattagat caatttaatt tatacgatag attgacagaa attcacgatg 300
aagtaacact tgaggatttt caaaaaacat taaatgatat ttacgatctg gtattaaatt 360
caaagtcatt ttctaatgta tcttctacaa tattcaataa aaataagaaa gttaattttg 420
atgaaaccaa aaaaggagat ttgtcacgta aatgtttgat gaatgctcga gattggggtg 480
ttttaccatt aatttctgtt gatgatgata ttgttacatg tggaacttta aagggaatac 540
tatcggaatg tcaatctcgt atattatctt ggaatgaatg taatctttct acgaaagaaa 600
catatagtga aaagaaaagt gaatatcaat ctattttaga tgattctatg accaaagatg 660
ctgatgttac tactgctatg atacaattca tggatgatgt ttcaaatgtg tatggttcaa 720
ataatgaaaa tcaattgaaa tggtttaaca atcgatttct aacatatgtt agaaataaaa 780
ttagaccatt tttattgaca aactcgccaa ttgacaattt tgaacaatcg gatacttctt 840
ataattgttc tattgaaatc gttagaattt taagtaaata tgaaatttta tggaaagatg 900
aagtttctgt taatagatat aagaaaacat gtgacgatgg aataaacatt gaaaaatata 960
gatatttggt acatgcaaaa tctgactttt tgaggtataa ggaaacagca tcttttaaag 1020
aaattcatgc tgtaaaaagt ccaatatcat tgtgttttgg aaacaattat caaccatttt 1080
cattgagtga tgttggtgac agacataata ttaactttgg ttataaattt ggtaaattag 1140
gaaaacagcg aaaggaatgt tcatttaatt taaattatag aaggaaaaaa gtaaaatatg 1200
caaacactcc cgttcggtct gacgaaaaca agtgttattt agataatctt gaaatcgaag 1260
atgcaaagaa tggttcttac aaactttcat atatggttaa taaaaaatat aaacgtgagt 1320
cgtttatcaa agaacctaaa atgaaaatgt ataatggtaa attgtatatg tattttccaa 1380
tgtcaaatga atttgaagaa gacagagatt catttgcgtt actaacttat tttagcagat 1440
catccaactc aaaatctcaa attgatgaag ctagtaacat attgcaaaat agaaagattc 1500
gtgtgtgtgg tgtggatttg ggaataaacc caacgtttgc tttatctgta ttagagtata 1560
gtgataacaa gataaccgac actaacatcg gcatgaagca tgaagggtct tataataact 1620
tcagtgaaat cagaaaacaa ataaatgatg taactgacat gatttcctat ttaaagtcaa 1680
aatatgataa ttgtgaaaaa gactattctt ctaaaataga cgatcatatc aaaagtagat 1740
tgaatgaaga aatttcaaat ttttgtgatt tggtttctta taaaagaaat aaaaatacga 1800
ttatacgaaa agaaataaag aatgtagaaa aggaaattaa taagattaaa aattgtcgcc 1860
gtcacactct taaaaaagat ttaactgaaa attttggatg ggtttctgca ttaaatgagt 1920
ttatatcatt aaaacattca ttcaatgata tgggagaatc gtttgattca aaaacaaatc 1980
caagttatag ttattttgaa aagtggaaac gttatattga caatataaag gatgattcgc 2040
tgaaaacagt ttcacgggaa attttaaatt tctgtataga aaactcagtt gatttcattg 2100
cgttggaaga cttacaaact ttcgctccgt cggatgatag aacgaaaagt cacaataaac 2160
taactcagtt atggtgtttt ggaaaattaa aaaaatgttt agaagatatt gcttctatgt 2220
atggtattca tgtgtattca agcacagacc ctagaaacac atctgacacg cattttgaat 2280
ccaaaaattt tggatatcgt gatgagtcaa ataaacataa tttatgggtt aatgtagatg 2340
gtgaatatac tgtggttgat tccgatatta atgcttctaa aaatattgca aatcggtttt 2400
taacgcatca taaagattta aaacaattgc caatgattgg tgatgggact ttatttaaaa 2460
ttgatagttc ttcaaaacga aataaatctt ttgcggtaaa attaaatata cacaaaaacg 2520
tatacgagtt aatagatggt gaatttgtaa aatcaaacaa aaaaccaaac ggaacttcac 2580
gtaaacaaac tgcttatata catggtgata tgtttatcga ttcaatatca cataaaaata 2640
aaaaaatgtt cttgagagaa aatctaataa gaaacgggtt tatatcaaaa taaaaataaa 2700
acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc 2760
tctcctgagt aggacaaatt tgacagctag ctcagtccta ggtataatgc tagcgctgag 2820
tctaatatga tagtaaaata ttatatagtt tagacggtat aacaacttcg acgagctcta 2880
cagtctaata tgatagtaaa atattatata gtttagacgt gtcagaggtg agtgttaact 2940
atgtgaagtc taatatgata gtaaaatatt atatagttta gacaaataaa acgaaaggct 3000
cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt 3060
aggacaaat 3069
<210> 93
<211> 35
<212> DNA
<213> 人工序列
<220>
<223> PAM文库序列
<220>
<221> misc_feature
<222> (1)..(8)
<223> n = a或g或c或t
<400> 93
nnnnnnnngg tataacaact tcgacgagct ctaca 35
<210> 94
<211> 3093
<212> DNA
<213> 人工序列
<220>
<223> Cas12h.1***表达盒的核苷酸序列
<400> 94
tttacacttt atgcttccgg ctcgtatgtt aggaggtctt tatcatggcc ctgatccaga 60
gggccggcgt gctgaagacc aagtccgact tcccgaaggt gatcaaggac tggcacgact 120
ccctgctggc cgactacagg aagttcttcc cgatcatctt ctcctggtgc ccggagtacg 180
gctacaccac catccaggac aacaagccgg tgttcgtgtc cccggaggag aggatggagt 240
ccatcaggaa ggaggccaag gagcacctga acgaggtgct ggccttcggc aagatgatcg 300
gctccaaggg cgtgggcggc tcctcctcct acgccatctt ctacaagcac cacaagaaca 360
acgagaacgg cgcctacacc ccgtccaggg ccaagttcat gaaggagggc atccacaaca 420
ggagggtgga gctggtggac gtgctgatgc tgaacgccat cccggacgag gagtgggtga 480
agatcgccca ggaggtggtg ggctactccg aggagaggct gaagctgtac tggaacaagt 540
tcatcgccaa gagggtggtg tcccacgaca ggaagctggg caagatcgtg agggagaagt 600
acctggagcc gaagggcctg gtgtgcgccc agccggagaa ctccacctac tgcagggtgc 660
tgaccgagat catcaagagg cagctgcact cccagatcga gaagtccaag ttccacgagg 720
aggagctgaa gtccatcgag aagaccgtgt ccgagttcga ctccccgctg ctggacttca 780
tctgccagta cgccgaggag ctgaaccaga tcaactccgg cctgtccaag tacgtgatca 840
agaacgccgt gaaggaggtg atctccccgc cggagaagca gtccgagatc tacgtgcagt 900
cccaggtgct gtcccaggag aagtacaagc cgctggtgaa cgccaccatc aaggagatcc 960
tgtccggcta cgagcagtgg aaggtgaagt ccaggtacga gaacaggctg aagaacagga 1020
agtacgtgct gtacccgaag ctgtccgcca actacaagat cccgatcggc cagaactccc 1080
tgggcaagtt caagatcaac gtgtccgaga acggcgagat cgtgatcagg ctgaacgaca 1140
tggccgacgt ggtgtgcatg ccgtccaagt acttcttcaa cctgaagtcc tccccggtgg 1200
tggacaagaa gaagcagctg gtgggctacc agatctcctt caaccacaac tccaggagga 1260
aggagccgac cgagaagccg gacttcaacg gcatcgtgaa ggagatcggc ctgcagctga 1320
aggacgacgg caggttctac atcaccctgc cgtactgcat ggagtactcc aacgacaact 1380
tcgacctgat caggccgctg ctgacctcct ccccgaccga ggaccagatc aagaagatgc 1440
cgtccgagtt caacgtggtg ggcttcgacc tgaacctgtc catgccgctg ccgatcacca 1500
gggccatcgt gggcaagtcc gtgaagggcg agatcaacgt ggagtacctg ggccaggcca 1560
aggtgatcga gtccacccac ctgatctacg acaacaacag gtgcaaggtg ctgatcgcct 1620
acaagaggca gtgcgacctg atcaagaggg ccatcaggga gtggaagatc tgcaagggca 1680
agaacatcga catctccgag aagacctacg agtggctgga gtcccacacc aagaggtgga 1740
acccgtccag gcagccggag tccatgcagg acaggttctc cgtgtccaag atgaggatcc 1800
agatcctggt gaacaaggcc aagtccagga tcgccaagta caacgacaac tcctggaaga 1860
ccggccacgg caacgagtcc gagctgatca ggctgatcga cgccgacgac gcctacaact 1920
ccctggtgtc cacctacaac aggatccacc tgaagtccaa ccagttcatc tacgccctgc 1980
cgtccaagaa caactccagg tccaacaaga aggagtactg cctgaggagg atcgccgcca 2040
agatcgccag gtactgccac ctgcacaacg tgaacatctg catcggcgag aacctgtcct 2100
tccagcagga ctccgacaac atctccaagg acaactccct ggtgaggctg ttctcctcca 2160
agtccatcgc caactacatg aagctggcca tggagaagtt cggcatcgcc ttcatcgact 2220
ccgccgaccc gtccggcacc tccaagaccg acccggtgac cggcaacatc ggctacagga 2280
acaagttcga caagaggaag ctgcacgtga tcaggaacgg caactggggc tgggtggact 2340
ccgacatcgc cgcctccctg aacatcctga tcaggggcat caacaggtcc atcgtgccgt 2400
acaagttctt cgtgggcaag aagaagcagg agtccaagag gctgaaccac ttcctgaaca 2460
agatcttcgg caccaccaag gtgttcttct acgaggacca gttcggcttc gccaacccgt 2520
ccctgtccaa gaaggagggc gagaacctga tcgccaacca gtacctgtac tacagggagg 2580
gcaagttcgt gacccagaag atccacaggc agatcgagga cgacttcaag aagatcgact 2640
tctccaacac cccggaggtg aacctgatcc cgtccggcgt gaagctgaag aacttccagt 2700
tcgagtgaaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg 2760
tttgtcggtg aacgctctcc tgagtaggac aaatttgaca gctagctcag tcctaggtat 2820
aatgctagcg ctgaatgtgc tgcatggcaa cgctagatgc catgtgttcg caacggtata 2880
acaacttcga cgagctctac aatgtgctgc atggcaacgc tagatgccat gtgttcgcaa 2940
ctatttcagc ccaattgggt ttatatcatg tgctgcatgg caacgctaga tgccatgtgt 3000
tcgcaacaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt 3060
ttgtcggtga acgctctcct gagtaggaca aat 3093
<210> 95
<211> 2682
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 74所示融合蛋白的编码核酸序列
<400> 95
atgctgtaca ccatgaacgt gaagaccatc aagctgaagg tggacgccac caaggaggtg 60
gagtccaggc tgaccaagat gctgctggtg cacaacaaca tcggcaggga gatcatcaac 120
ttcctgatcc tgtgctccgg caacgacaac atcaggaaga ccaagttcga cgagttcggc 180
aactcctacg acgagttctg caacctgaag ctggaccagt tcaacctgta cgacaggctg 240
accgagatcc acgacgaggt gaccctggag gacttccaga agaccctgaa cgacatctac 300
gacctggtgc tgaactccaa gtccttctcc aacgtgtcct ccaccatctt caacaagaac 360
aagaaggtga acttcgacga gaccaagaag ggcgacctgt ccaggaagtg cctgatgaac 420
gccagggact ggggcgtgct gccgctgatc tccgtggacg acgacatcgt gacctgcggc 480
accctgaagg gcatcctgtc cgagtgccag tccaggatcc tgtcctggaa cgagtgcaac 540
ctgtccacca aggagaccta ctccgagaag aagtccgagt accagtccat cctggacgac 600
tccatgacca aggacgccga cgtgaccacc gccatgatcc agttcatgga cgacgtgtcc 660
aacgtgtacg gctccaacaa cgagaaccag ctgaagtggt tcaacaacag gttcctgacc 720
tacgtgagga acaagatcag gccgttcctg ctgaccaact ccccgatcga caacttcgag 780
cagtccgaca cctcctacaa ctgctccatc gagatcgtga ggatcctgtc caagtacgag 840
atcctgtgga aggacgaggt gtccgtgaac aggtacaaga agacctgcga cgacggcatc 900
aacatcgaga agtacaggta cctggtgcac gccaagtccg acttcctgag gtacaaggag 960
accgcctcct tcaaggagat ccacgccgtg aagtccccga tctccctgtg cttcggcaac 1020
aactaccagc cgttctccct gtccgacgtg ggcgacaggc acaacatcaa cttcggctac 1080
aagttcggca agctgggcaa gcagaggaag gagtgctcct tcaacctgaa ctacaggagg 1140
aagaaggtga agtacgccaa caccccggtg aggtccgacg agaacaagtg ctacctggac 1200
aacctggaga tcgaggacgc caagaacggc tcctacaagc tgtcctacat ggtgaacaag 1260
aagtacaaga gggagtcctt catcaaggag ccgaagatga agatgtacaa cggcaagctg 1320
tacatgtact tcccgatgtc caacgagttc gaggaggaca gggactcctt cgccctgctg 1380
acctacttct ccaggtcctc caactccaag tcccagatcg acgaggcctc caacatcctg 1440
cagaacagga agatcagggt gtgcggcgtg gacctgggca tcaacccgac cttcgccctg 1500
tccgtgctgg agtactccga caacaagatc accgacacca acatcggcat gaagcacgag 1560
ggctcctaca acaacttctc cgagatcagg aagcagatca acgacgtgac cgacatgatc 1620
tcctacctga agtccaagta cgacaactgc gagaaggact actcctccaa gatcgacgac 1680
cacatcaagt ccaggctgaa cgaggagatc tccaacttct gcgacctggt gtcctacaag 1740
aggaacaaga acaccatcat caggaaggag atcaagaacg tggagaagga gatcaacaag 1800
atcaagaact gcaggaggca caccctgaag aaggacctga ccgagaactt cggctgggtg 1860
tccgccctga acgagttcat ctccctgaag cactccttca acgacatggg cgagtccttc 1920
gactccaaga ccaacccgtc ctactcctac ttcgagaagt ggaagaggta catcgacaac 1980
atcaaggacg actccctgaa gaccgtgtcc agggagatcc tgaacttctg catcgagaac 2040
tccgtggact tcatcgccct ggaggacctg cagaccttcg ccccgtccga cgacaggacc 2100
aagtcccaca acaagctgac ccagctgtgg tgcttcggca agctgaagaa gtgcctggag 2160
gacatcgcct ccatgtacgg catccacgtg tactcctcca ccgacccgag gaacacctcc 2220
gacacccact tcgagtccaa gaacttcggc tacagggacg agtccaacaa gcacaacctg 2280
tgggtgaacg tggacggcga gtacaccgtg gtggactccg acatcaacgc ctccaagaac 2340
atcgccaaca ggttcctgac ccaccacaag gacctgaagc agctgccgat gatcggcgac 2400
ggcaccctgt tcaagatcga ctcctcctcc aagaggaaca agtccttcgc cgtgaagctg 2460
aacatccaca agaacgtgta cgagctgatc gacggcgagt tcgtgaagtc caacaagaag 2520
ccgaacggca cctccaggaa gcagaccgcc tacatccacg gcgacatgtt catcgactcc 2580
atctcccaca agaacaagaa gatgttcctg agggagaacc tgatcaggaa cggcttcatc 2640
tccaagtcca gggccgaccc gaagaagaag aggaaggtgt ga 2682
<210> 96
<211> 2841
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 75所示融合蛋白的编码核酸序列
<400> 96
atgaacaaga ccgacaccca gaacaacgag cagatcaaca agccgaccca gctgctgaac 60
aacaaggaca tcgagctgac cgtgaagacc gtgaagtccg ccaccgtgaa ggtggacaac 120
aactccaaga aggagctgtt cggcctgttc aactacttca cctccgtggc ctccggcatc 180
aaggacaagg tgtacaacct gcagtccgac gagaagaccg ccccgatctt caacgactac 240
gtgaagcagc cgcagagggg caggtccgcc gccaccaccc tgttcaccaa gctggacgcc 300
gagaagacct acacctccca gcactccttc ccgggcaagt ggagggactc cggcatcttc 360
ccgctgtaca acaaggagtc cgagaagtac gacctgtcca cccacggcta ccactactcc 420
gccaacgccg agatccacac ccagctggac tcccacgacg agtgcaacaa ggagtgcgag 480
aaggagtacg ccgccctgag ggacgaggtg aacaactaca agtacgagtt caccctgcag 540
ttcaaggccg agaacgccga gaagttctac aacttcgtgg agaagctgac cctgatgggc 600
tggaggtacg acgccacctt caggtccttc ttcgagctgc acatgcaccc gaagctgaag 660
accggcgaga ccacctacag ggccacctac aagctgccgt ccggcaagtc caagaggtac 720
tccttcttca gggacgacat cgccgacgag atcgccaaga acccggagtt ctggccgatg 780
ctggagtcct ccaacgccat ctcctggatc aactccaaca acctgctgtc caggaagaag 840
gacaaggcca actactcctc cacctccctg atcaagtccc agatcaggct gtacctgggc 900
aacaacggcg tgccgttcac cgccagggag cacgacggca ggatctactt ctccttcagg 960
ctgccggcca tcaacggcga gaagggcagg atggtggaga tcccgtgctc ctacaagaag 1020
gtgttcaacg gcaaggccag gaagtcctgc tacctgggcg gcctgaccat cgagaagacc 1080
gacgccggca agcacatctt caagtactcc gtgaacaaca agaagccgca ggtggccgag 1140
ctgaacgagt gcttcctgag gctggtggtg aggaacaggg agtacttcaa caacgtggtg 1200
gccggcaaga tcaccgacat caacaccgac cacttcgact tctacgtgga cctgccgctg 1260
aacgtgaagg aggacccgat ccacgacctg tcctccaccg aggtgttcgg caagaacggc 1320
ctgaggtcct actactcctc cgcctacccg gagatcaaga acctgggctc ccagatcgag 1380
accggcaaga acctgacctg cccgatcacc aagacccaca acatcatggg catcgacctg 1440
ggccagagga acccgttcgc ctactgcatc aaggacaaca ccggcaagct gatcgcccag 1500
ggccacatgg acggctccaa gaacgagacc tacaagaagt acatcaactt cggcaaggag 1560
tccacctccg tgtcccacct gatcaaggag accaggtcct acctgcacgg cgacccggag 1620
gccatctcca aggagctgta caacgaggtg gccggcttct gcaacaaccc ggtgtcctac 1680
gaggagtacc tgaagtacct ggactccaag aagttcctga tcaacaagga ggacctgtcc 1740
aagaacgcca tgcacctgct gaggcagaag gaccacaact ggatcggcag ggactggctg 1800
tggtacatct ccaagcagta caagaagcac aacgagaaca ggatgcagga cgccgactgg 1860
aggcagaccc tgtactggat cgactccctg tacaggtaca tcgacgtgat gaagtccttc 1920
cacaacttcg gctccttcta cgacaagaac ctgaagaaga aggtgaacgg caccgtggtg 1980
ggcttctgca agaccgtgca cgaccagatc aacaacaaca acgacgacat gttcaagaag 2040
ttcaccaacg agctgatgtc cgtgatcagg gagcacaagg tgtccgtggt ggccctggag 2100
aagatggact ccatgctggg cgacaagtcc aggcacacct tcgagaacag gaactacaac 2160
ctgtggccgg tgggccagct gaagaccttc atggagggca agctggagtc cttcaacgtg 2220
gccctgatcg agatcgacga gaggaacacc tcccaggtgt gcaaggagaa ctggtcctac 2280
agggaggccg acgacctgta ctacgtgacc gacggcgagt cccacaaggt gcacgccgac 2340
gagaacgccg ccaacaacat cgtggacagg tgcatctcca ggcacaccaa catgttctcc 2400
ctgcacatgg tgaacccgaa ggacgactac tacgtgccga cctgcatctg ggacaccacc 2460
gaggagtccg gcaagagggt gaggggcttc ctgaccaagc tgtacaagaa ctccgacgtg 2520
gtgttcacca agaagggcga caagctggtg aagtccaaga cctccgtgaa ggagctgaag 2580
aagctggtgg gcaagaccaa ggagaagagg ggccagtact ggtacaggtt cgagggcaag 2640
tcctggatca acgaggccga cagggacacc atcatcctga acgccaagaa gatctccagg 2700
gagagggaca acggcgagca gtccaccgac accaggtccc agaacgtgac cgtgtccgtg 2760
ctggacgtgt gcgagaccgc cgagaagaag aagctggtgc tggtgtccag ggccgacccg 2820
aagaagaaga ggaaggtgtg a 2841
<210> 97
<211> 2697
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 76所示融合蛋白的编码核酸序列
<400> 97
atggccctga tccagagggc cggcgtgctg aagaccaagt ccgacttccc gaaggtgatc 60
aaggactggc acgactccct gctggccgac tacaggaagt tcttcccgat catcttctcc 120
tggtgcccgg agtacggcta caccaccatc caggacaaca agccggtgtt cgtgtccccg 180
gaggagagga tggagtccat caggaaggag gccaaggagc acctgaacga ggtgctggcc 240
ttcggcaaga tgatcggctc caagggcgtg ggcggctcct cctcctacgc catcttctac 300
aagcaccaca agaacaacga gaacggcgcc tacaccccgt ccagggccaa gttcatgaag 360
gagggcatcc acaacaggag ggtggagctg gtggacgtgc tgatgctgaa cgccatcccg 420
gacgaggagt gggtgaagat cgcccaggag gtggtgggct actccgagga gaggctgaag 480
ctgtactgga acaagttcat cgccaagagg gtggtgtccc acgacaggaa gctgggcaag 540
atcgtgaggg agaagtacct ggagccgaag ggcctggtgt gcgcccagcc ggagaactcc 600
acctactgca gggtgctgac cgagatcatc aagaggcagc tgcactccca gatcgagaag 660
tccaagttcc acgaggagga gctgaagtcc atcgagaaga ccgtgtccga gttcgactcc 720
ccgctgctgg acttcatctg ccagtacgcc gaggagctga accagatcaa ctccggcctg 780
tccaagtacg tgatcaagaa cgccgtgaag gaggtgatct ccccgccgga gaagcagtcc 840
gagatctacg tgcagtccca ggtgctgtcc caggagaagt acaagccgct ggtgaacgcc 900
accatcaagg agatcctgtc cggctacgag cagtggaagg tgaagtccag gtacgagaac 960
aggctgaaga acaggaagta cgtgctgtac ccgaagctgt ccgccaacta caagatcccg 1020
atcggccaga actccctggg caagttcaag atcaacgtgt ccgagaacgg cgagatcgtg 1080
atcaggctga acgacatggc cgacgtggtg tgcatgccgt ccaagtactt cttcaacctg 1140
aagtcctccc cggtggtgga caagaagaag cagctggtgg gctaccagat ctccttcaac 1200
cacaactcca ggaggaagga gccgaccgag aagccggact tcaacggcat cgtgaaggag 1260
atcggcctgc agctgaagga cgacggcagg ttctacatca ccctgccgta ctgcatggag 1320
tactccaacg acaacttcga cctgatcagg ccgctgctga cctcctcccc gaccgaggac 1380
cagatcaaga agatgccgtc cgagttcaac gtggtgggct tcgacctgaa cctgtccatg 1440
ccgctgccga tcaccagggc catcgtgggc aagtccgtga agggcgagat caacgtggag 1500
tacctgggcc aggccaaggt gatcgagtcc acccacctga tctacgacaa caacaggtgc 1560
aaggtgctga tcgcctacaa gaggcagtgc gacctgatca agagggccat cagggagtgg 1620
aagatctgca agggcaagaa catcgacatc tccgagaaga cctacgagtg gctggagtcc 1680
cacaccaaga ggtggaaccc gtccaggcag ccggagtcca tgcaggacag gttctccgtg 1740
tccaagatga ggatccagat cctggtgaac aaggccaagt ccaggatcgc caagtacaac 1800
gacaactcct ggaagaccgg ccacggcaac gagtccgagc tgatcaggct gatcgacgcc 1860
gacgacgcct acaactccct ggtgtccacc tacaacagga tccacctgaa gtccaaccag 1920
ttcatctacg ccctgccgtc caagaacaac tccaggtcca acaagaagga gtactgcctg 1980
aggaggatcg ccgccaagat cgccaggtac tgccacctgc acaacgtgaa catctgcatc 2040
ggcgagaacc tgtccttcca gcaggactcc gacaacatct ccaaggacaa ctccctggtg 2100
aggctgttct cctccaagtc catcgccaac tacatgaagc tggccatgga gaagttcggc 2160
atcgccttca tcgactccgc cgacccgtcc ggcacctcca agaccgaccc ggtgaccggc 2220
aacatcggct acaggaacaa gttcgacaag aggaagctgc acgtgatcag gaacggcaac 2280
tggggctggg tggactccga catcgccgcc tccctgaaca tcctgatcag gggcatcaac 2340
aggtccatcg tgccgtacaa gttcttcgtg ggcaagaaga agcaggagtc caagaggctg 2400
aaccacttcc tgaacaagat cttcggcacc accaaggtgt tcttctacga ggaccagttc 2460
ggcttcgcca acccgtccct gtccaagaag gagggcgaga acctgatcgc caaccagtac 2520
ctgtactaca gggagggcaa gttcgtgacc cagaagatcc acaggcagat cgaggacgac 2580
ttcaagaaga tcgacttctc caacaccccg gaggtgaacc tgatcccgtc cggcgtgaag 2640
ctgaagaact tccagttcga gtccagggcc gacccgaaga agaagaggaa ggtgtga 2697
<210> 98
<211> 2799
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 77所示融合蛋白的编码核酸序列
<400> 98
atggccacca ggtccttcat caggaccggc aacctgaagg ccaagaacac cgccgaggag 60
gtgatgcagt ggtacgccga cctgcagtcc gactacaggt ccttcctgaa cctgttcttc 120
ggctggatgg ccatcggcta cggcaccaac gccgaggacg aggtgttcta cacctccaag 180
gaggagtccg agaggctgag gtccctgacc atcggcgacg ccaagaagga gcagctggcc 240
gtgtccttca tcgagctgct gctgaagggc ggcgagaacg cctcctcctg ctacaacgtg 300
ttctacagga actacaagtc cctgggcaag gccaagctga cccagaagaa gaacgacttc 360
ctgtccgccc tgccgctgct ggacgagaac aagatcaagg agtacttcaa gaccgacgag 420
cagctgtccc agatctgcat cgaggagtgg ctggagtacg gcgtgaagaa cctgccgctg 480
ccggagatct gggccgaggt gtccccgagg ctggcctcca tcgagaggtc cctgggcgtg 540
gacctgaggc tggccttcgg cctgtcctgc atcaggtcca gggactgcaa ctactgcagg 600
atcctgatcg agatggtggg cagggacctg aggtccatct tcgagaagta caacaaccac 660
ctgctggaga ccgagaagat caagctgtcc atgaacgaca agcagggccc ggtgtacgac 720
tccatctgct gcttcgccgc cgagctggag tccaagaact ccggcctgac caagtacgtg 780
ctgaccaagg gcatcgacca cgtgaagaag ggcaccggcg agaagaccga catcaggctg 840
gccgtgaagg agctgaagaa gaacaagtac aggatcctga tcgagtcctc ctactccgag 900
atcatgtccg cctactcctg ctggaggacc aagaagcagc tggagaagag gaagctgtac 960
ccgtgcttcg acccgaacag gaacgactac aaggtgccgg tgggccaggg ctccctgggc 1020
aacttcaccg tgtccgtgga ggactccggc gacgtgctga tcgagatcgt gggcgtgggc 1080
gtgatcaggt gcgccgcctc ctgctacttc tccggcatcg tgttcgacga gatcaggaac 1140
aagaacggca ggaccggcta ctccctgaac ttctgccaca agtccatctc caagggcaag 1200
aaggccgtga aggccgcctc ccacaccggc gacaagatct ccggcgtgct gaaggagatc 1260
ggcctgagga acaccgactc cggcttcttc gtgtccctgc cgtactccat ccaccacgac 1320
gagaagaact tcaagatcgc cgagttcttc atgtccgcct gcccgaagaa ggagaacgtg 1380
gagaacctgc cggacaagat cgtggtgggc gccatcgacc tgaacgtgtc caacccggtg 1440
gccgccgtga aggccgtggt gtacagggac gacaagtccg gccagctgaa cgccctggac 1500
tacggctccg gcaacctgat caagaagccg ttcatgctgg tggccaacgg cccgaggatc 1560
aagaacctga tcgagatcag ggacgacgcc aggagggtga tcggcgccat cagggagttc 1620
aaggtgtcca acgccgtgaa ggagcacgtg ggcgaggaca ccagggactt cctgatcctg 1680
tgcggcgaca ccaagtcctc ctccaccagg tacctgatcc agtcctgggt gaagaagatc 1740
aactccaggc tgaggaagat caagttcgag atgaggtccg gcggctacag ggactgcgcc 1800
gacaacatca ggctgatcga ggccatggac cagtgcgcct ccatggccga gtcctacaac 1860
aggatccacc tgaagtccgg cgagaagctg gtgaaggtgg ccaagttcga caagtccagg 1920
gccaacttca ggaacttcgt gctgaggcag ctggcctcca agatcgccaa cgagatgaag 1980
gactgcaacg tggtgttcgg cgaggacctg gacttcatct tcgactccga caagaacaac 2040
aacgccctgc tgaggctgtt ctccgccgcc accctgctga agtacatcat cgaggccctg 2100
gagaagatcg gcgtgggctt cgtgaaggtg gccaagaacg gcacctccca gtccgacccg 2160
gtgacctcca acccgggctg gagggacgac aagaacaagt ccaggctgta cgtggtgagg 2220
gacaagcagc tgggctggat cgactccgac ctggccgcca ccatgaacat cctgatccag 2280
ggcctgaacc actccgtgtg cccgtacaag ttctacgtga aggagtacga gaacaagccg 2340
aactccaccc aggactccat caacgccatc aagaagccgg aggaggccat cggcaagagg 2400
atcaagaggt tcttcaacct gaagtacggc tcctccgtgc cgaagttcgt gtccgacgac 2460
aggggcaggg tgaccttcgc caagaagatc gactccaccc agaccaggct gatcaaccag 2520
ttcgtgtacg cccactcctc ctgcatcgtg acctgcgagc tgcacaacga gatggtgaac 2580
aagatcaagc agctggccgt ggagaagccg aactgccagg agttcgacgt gacctgcgac 2640
ccggacggca ggtacaacaa cttcgccctg ccggaggtgc acgactcctc caaggacgtg 2700
ggcgccaagg ccctgaccac caaggacgtg gacttcaaga ccatcctgaa ggaccacacc 2760
gcctccaggg ccgacccgaa gaagaagagg aaggtgtga 2799
<210> 99
<211> 3927
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 78所示融合蛋白的编码核酸序列
<400> 99
atgggcaaga acgagaacaa gtaccagctg tccaagaccc tgaggttcgg cctgaccctg 60
aaggagaaga tctccaacaa cgagaagacc ccgtaccagt cccactccca gttcagggac 120
ctgatcatcc tgtccgagaa caggatcagg gagggcatct ccaccccgca gaacagggac 180
ctgccgtcct tcatccacag gatccagaac tgcaccgact tcatcaacga cttcatccac 240
gactggtgga tgatcctgat gcacaccggc cagatcgagc tggacaagga ctactacaag 300
tccctgacca agaaggtggg cttcgtgggc ttctggtaca aggagaacaa gaagaagggc 360
ggcaagacca agcagccgca ggccaggaac atcccgatgg gcgagctgag gcacctgtgc 420
ccgcagaaca ccaaggagtg cgccacctac atcaccgact actggaagga cctgctgatc 480
accgccacca acaagctgta cgagtcctcc gagcagcaga agaagttcat caaggccatg 540
gagcagaaca ggaccgacaa caagccgaac gagatcgacc tgaagaagtc cttcctgtcc 600
ctggtgtccg tgaccatgga gctgctgaac ccgatcctga acggccagat cctgttcaac 660
aagatggaca ggctggacat gtccaagaag tccgacaacg acttcatcga cttcgtgaac 720
gaccacgaga ccgtgaggga gctgaacaac gacatcgagg agatcatcgc cgacttcaag 780
gagaacggca acaacgtgaa ctactgcaag gccaccctga acccggacac cgccctgaag 840
cagcacaaca acaacatccc gaacgacatc gccaccgacc tggaggagct gatgatggac 900
tccatcgtgg gcaactacga cgacgtgaac tccttcatgg acaactacgt gtccaacctg 960
tccgccaagg acaagatcaa gaagatcaag gactccaaca tctccctgat ctacagggcc 1020
atcctgttca agtacaagat gatcccggcc aacgtgagga gggacatcgc ccagggcatg 1080
gccaagaagc tgaacaagga cgaggagaac atctactcct tcctgtgcga gttcggcacc 1140
ctgaggaccc cgcagaagga ctacgccgac ctgaaggaca aggactcctt caacctggac 1200
aactacccgc tgaaggtggc cttcgacttc gcctgggagg gcctggccaa ggcctggtac 1260
cacgaccagt ccgacttccc gatcgacccg tgcagggact tcctgcagga gaacttcgac 1320
gtgaacctgg aggaggacca ggaggacgag tacttcctgc tgtacgccga cctgatcgag 1380
ctgaacgccc tgctgtccac cctggacaag ggcaacccgg ccgacccgga ctccatcaag 1440
aacgaggccc tggagatggt ggagtacatc aactggaact ccctggacaa gaagaacggc 1500
aactactaca agaagatcat caagaacagg ctgaagtcct ccaagggcaa cgagacctac 1560
gagaggatca agaaggagat ctccatgtcc aggggcaggc tgaagaacaa gatcgagaag 1620
tacgacgacc tgacctccca gtacaagagg atcgccatgg acctgggcaa gaagttcgcc 1680
tccctgaggg acaagatcat cgccgccaac gaggacaaca aggtgaccca ctacgccatg 1740
atcctggagg actccaactg cgacaagtac ctgctgctgc agaaggtgtc caacaacatc 1800
taccactgca tgtcctacga ctcctccgac ccgaaggcct actacgtgga ctccatcacc 1860
tcctccgcca tcgccaagat gatcaggaag gagaccaacc cgtccaagat cagggagtac 1920
gccgagctgg aggagaagga gagggagagg aggaacgtgg acgactggtg caggttcatc 1980
tccaagaagg agtacgacag gaggtaccag ctgaacatca acaacggcct gtccttcgag 2040
gccctgaaga aggagatcga ctccaagtcc tacatcctgg tgaagaagaa catctccgtg 2100
gactccatca gggagctggt ggagaacgag ggctgcctgc tgttcccgat cgtgaacaag 2160
gacctgacca aggagaggaa gaccaccgag gacaaccagt tcaccaagga ctggaacatg 2220
atcttctccg gctccgagac caactggagg ctgaccccgg agttcagggt gacctacagg 2280
aacccggtgc cgggctaccc gaacgacaag ttcggctcca agaggtactc caggttccag 2340
atgaacgccc acttcgtgtg cgacttcatc ccgtcctcca actcctacac ctccaacagg 2400
gagcagatcg ccatcttcaa ggacgagggc gagcagaaga agagggtgga ggagttcaac 2460
aggaccctgt ccaacatcaa ccagaagttc tacgtgatcg gcatcgacag gggccagaag 2520
gagctggcca ccctgtgcgt ggtggaccag gacaagaaga tccacggcga cttcaagatc 2580
tacaccagga agttcaactc cgagaggaag cagtgggagc actactccct ggagggcgag 2640
aagggcacca ggaacatcct ggacctgtcc aacctgaggg tggagaccac catcatcatc 2700
gacggcaagc cggagaggag gcaggtgctg gtggacctgt ccgaggtgct ggtgaaggac 2760
aaggagggca actacaccaa gccgaacaag atgcagatca agatgcagca gatggcctac 2820
gtgaggaagc tgcagttcca gatgcaggcc aacccgaccg aggtgctgga gtggtacgag 2880
cagaacccga ccgaggagct gatcatcaag aacctggtgg acaaggagaa cggcgagaag 2940
ggcctgatct ccttctacgg caccgccctg gtggagctgg accagaccct gccggtgtcc 3000
aagatcaagg agatgctgga ggagttcaag atcctgaagc agagggagtc caagaaggag 3060
aacgtgcaga aggagctgaa caacctgacc cagctggagg ccgtggactc cctgaaggcc 3120
ggcatcgtgg ccaacatggt gggcgtgatc tcctacatcc tgaagaccct ggactacaac 3180
gcctacatct ccctggagga cctgtccacc gtgcagtcct ccaccgagtt cgcctccggc 3240
atctccggcg ccatcaccaa gatgtccagg gaggagggca ggaggatcga cgtggagaag 3300
tacgccggcc tgggcctgta caacttcttc gagatgcagc tgctgaggaa gctgcacagg 3360
atccagaccg acaacggcaa catcctgcac ctggtgccgg ccttcagggc ccagaagaac 3420
tacgaccaca tcatggtggg caaggagaag atcaagaacc agttcggcat cgtgttcttc 3480
gtggacgccg ccgccacctc catcaagtgc ccgaggtgcg gcgccgtgaa cgaggacaag 3540
ttcaacccgg acaagcagaa gtacccggac gccgagaagg gcccgaagct gaggaacagg 3600
aaggagcagt ccggcaagaa ggtgtgggtg accagggaca aggaggacga cgacaggatc 3660
aagtgctact gctgcggctt cgacaccaag gagaagaacg agggcaaccc gttcatgtac 3720
atcaagtccg gcgacgacaa cgccgcctac ctgatctccg acctgggcgt ggagtcctac 3780
aggaaggcct acgagctggc cgccaccgtg gtggaggaca ggaagaagac cctgaccaac 3840
aacctgaacc agtccaacta caagatcagg ttcctgtggc acaccatgta ctccagggcc 3900
gacccgaaga agaagaggaa ggtgtga 3927
<210> 100
<211> 3807
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 79所示融合蛋白的编码核酸序列
<400> 100
atggacgccg acaagaccac caaggccatc aacgagtacc agacccagaa gaccatcagg 60
ttcggcctga ccgccaccaa ccagaacctg tactccgagg agatcatgaa gctgctgaac 120
atctccgagg agaggatcat caaggagaag gtgaaggtga acaacgacac cgacaagacc 180
aaccagctga ggggctgcct ggtgcagatc aagaagtacc tgaagacctg ggagaacatc 240
tacgcccaga tcgacttcct ggccatcacc aaggactact acaaggtgat ctccaagaag 300
gccaggttcg acttcgacaa gggcaacggc tccgagatca agctgtcctc cctgcagtcc 360
acccacaaca agaagaagag gtaccagtac atcatcgact tctggaagga gaacctgagg 420
aagaccgaga acctgtacag gaagtccgac gacctgctga agatcttcga ggaggccaag 480
aaccagaaca gggacgacaa gaagctgaac aaggtggagc tgaggaagac cttcctgaac 540
ctgttcaccc tggtgaacga gtccctgaag ccgctgatcg agggcaacct gttcatcgtg 600
aacgacgaca agatcgacga gaagaactcc aagcacaact acgtgttcta cttcatctcc 660
aagaccgagg agaggaggct gctgtacgac aacatctgca ccctgcagga ctacttcaag 720
aacaacggcg gctacgtgcc gttcggcagg gtgaccctga acaagtggac cgccctgcag 780
aagttcaaca acagggacat cgagatcaac aggatcatca aggagctgaa gatcaacaac 840
atctccaccc agaagaccga ctacaagtac aacgacttca ccgagaactt caaggagaag 900
aaggacgaga acggcaaggt ggtgaagaac tccgccggca acatcatctg ggacctgaag 960
gccaacgcca agtccgtgat cgagatctgc cagttcttca agtacaagaa ggtgccgatc 1020
aacgccaggc tgaacctggc caagaggctg atcaaggaca acaagctgaa gaaggagcag 1080
gagaacacct tcctgtccga gttcggcgtg ctgaagaccc cggccttcga ctacgccagg 1140
gacaaggaga acttcaacct gaccaactac ccgctgaagg tggccttcga ctacgcctgg 1200
gagaactgcg ccaaggacaa gtacgagaag atcccgttcc cgaaggagca gtgcgagagg 1260
tacctgcaga ccgccttcga gatcgacgcc accaaggacg agaacaagaa gctgatcgac 1320
acccacctga acaagtacgc cgacctgctg cagttcaaga tcctgctgga gaggttcaag 1380
gccgagttcc acaagaccaa cgaggagacc aacaagaaca acatccagaa gctgaggaac 1440
gtgttctccg gcctggacta ccacggcgac aacaggctga acaagaacca gatccagaag 1500
gccatcgagg cctggttcga caacaaggag cagaacatcg gcaagaagaa ggagaacgag 1560
aagctgctga ccgagaacga gaagaacaac ttctccctgt ccatgcagat catcggccag 1620
gagaggggcg gcctgaagaa cggcatcccg aagtacaagg agctgaccga gatgttcaag 1680
gtgtgcgcct ccaagttcgg caagcagttc gccgacctga gggactactt caacgaggcc 1740
tacgaggtgg acaagatcaa gtacagggcc tggatcatcg aggacgacaa gaagaacagg 1800
ttcgtgctgt tcgtgaacaa ggagaaggcc ttcgacctga cctccgagga gggcgacctg 1860
tggttctacg aggtgaagtc cctgacctcc aagtccctgg tgaagttcat caagaacagg 1920
ggcgcctacc cggacttcca cgacgtgaag aactccttcc actactcctc catcaagaag 1980
gactggcaga actacaagaa cgacccggag ttcctggaca agctgaagga gtgcctgaag 2040
aactccaaga tcgccaagga ccagaagtgg gccaagttct gctgggactt caagcagtgc 2100
gacacctacg agaagctgga gaaggaggtg gacaggaagg gctacaagct ggagggctgc 2160
aagtccgagc cgaagaccat ctccctgacc cagctgaccg actgggtgga gaacaaggac 2220
tgcttcctgc tgccgatcgt gaaccaggac atcaacaagg gcgacaagag gaccaagaac 2280
cagaaccagt tcaccaagga ctggttcgac atcttcgaga acaagaagag gctgcacccg 2340
gagttcaaca tcttctacag gttcccgacc aaggactacc cgaacaccaa gttcaagaac 2400
ggcaccgaga agaccaagag gtactccagg ttccagatgc tggcctactt cggctgcgag 2460
gtgatcccgt ccggcaacca cctgtccaag aaggagcaga tcgccatctt caacaacgac 2520
aagaagcaga aggaggaggt ggagaagtac aacaagtcca tctcctccga ctgcgactac 2580
gtgatcggca tcgacagggg catcaagcag ctggccaccc tgtgcgtgct ggacaagaac 2640
ggcgtgatcc agggcgactt ccagatcttc accaggacct tcaacaagca gaccaagcag 2700
tgggagcaca aggagctgga gcagaggaac atcctggacc tgtccaacct gagggtggag 2760
accaccatca ccggcaagaa ggtgctggtg gacctgtcca agatcaagga cgacgagggc 2820
aactacacca acctgaagca gaccatcaag ctgaagcagc tggcctacat cagggagctg 2880
cagtacgcca tgcagaccag gccggacgac ctgctggact tcgtgaagtc catcaactcc 2940
gccaacgaca tcaccgccga gaacatcaag cacttcatct ccccgtacaa ggagggcaag 3000
aactacgacg acctgccgaa ggtggagatg ttcaacctgc tgaaggagtg gggcaacgcc 3060
gacgagaacg gcaagaggaa gatcgccgag ctggacccgg ccgacaacct gaagtccggc 3120
atcgtggcca acatggtggg cgtggtggcc ttcctgtgcg agaactacaa ctacaaggtg 3180
aggatcgccc tggaggacct gaccagggcc tacggcatcc agaaggacgc cctgaacggc 3240
accgccatct accagaacga cgaggacttc aaggagcagg agaacaggag gctggccggc 3300
gtgggcacca tgcagttctt cgaggtgcag ctgctgagga agctgttcaa gatccaggtg 3360
gacaagaacc tgcacctgat cccggccttc aggtccgtgg acaactacga gaagatcgtg 3420
aggagggaca agcagaactc cggcgacgag ttcgtgaact acccgttcgg catcgtgtgc 3480
ttcgtggacc cgaagtacac ctcccagcag tgcccgtact gcaacaacac ccacaagcac 3540
aagaagaacg acaccgagac cggcaagaag gccttctaca ggaacaaggg cgagaacaag 3600
aactccctgc tgtgcgagaa gtgcggcgtg tccaccatcg agggcgagga gaccctgtcc 3660
tccaagaacg acaacaagaa gcagttcaac atccactaca tcaccgacgg cgaccagaac 3720
ggcgcctacc acatcgccaa caaggtggtg atcaacttcc agaaggactc ctccagggcc 3780
gacccgaaga agaagaggaa ggtgtga 3807
<210> 101
<211> 3069
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 80所示融合蛋白的编码核酸序列
<400> 101
atggactacc agcagtacga gttcaccagg accatcaggt tcaacctgtc cggcgacgac 60
aagagggccc tgatgctgga cctgctggac gacacccagg agggcatgct ggccgccttc 120
caggagacct acaagaacct gctgttcgcc ttccaggagg ccatcctgag ggccgacggc 180
tccggcaacc tgagggtggg caggctggag atcaagaagt cctggctgag gcagtacgcc 240
agggagtact tctacgccct gtccgaggac gagaggaggt gcaagaacaa gttccaggcc 300
aagctgttcg acagggtgct gtccgactgg ctggagagga acaacgagct gctgcagagg 360
ctgaacaaca tcctgtccct gccgcaggag tccaagaccg gcgcctccga cctgtccctg 420
ctggtgaggc agctgaaggg cgccgagtac ttctacttca tcagggactt cacccagtcc 480
ggcatcatca acgacaagga ctccgacgag cacatcaaga acctggccgg catcgtggag 540
aagttcgaga ccctgctgga caaggtgctg ttcctgaccg ccccgaactc ctcccagggc 600
gtggagacca ccagggcctc cttcaactac tacaccgtga acaagatctc caagaacttc 660
gacgagaaca tcaagaaggc caacggcagg ctgtgctcct cctaccagaa ctccatgaac 720
gaggagctgc tgaggaaggt gggcttcctg aagtacctga aggacgagta cagggccgag 780
ctgcagaacg tgtccctgaa ggacctgtac gaggccctga agaagttcaa gtcccagcag 840
aagaccgcct tcatccaggc cgtgcagaag aacaagtccg agaaggagct gatgagggag 900
ttcccgctgt tcaacggcaa gcagccggac accctgcaga agttcatcct ggagaccgac 960
aagatcaaga ggggcgccta cttccagaag tggggcttcg acaactacat ctccttctgc 1020
aacaagatct tcaagccggt ggccatggag accggcacca ggaaggccaa gatcagggcc 1080
ctggagcagg agaagatcga ggccaggctg ctgcagtact gggcccacat cctggtgaag 1140
gacggcaagt acttcctgct gctgatcccg aaggagaaga tgggcgaggc caaggtgttc 1200
ttcgccaggc tgtccgacca ggagggcggc gagtacaccc tgtacgcctt caactccctg 1260
accctgaggg ccctgaagaa gctgatcagg aggaacctgg gcaaggagca ggtgaggctg 1320
tccgccggcg acgccgacgc catcgccctg tgccaggagg tgctgagggg caggtaccac 1380
cagctgaagg acctggacct gtccggcttc gagaaggaga tcgccgagat cgccaacacc 1440
cagtacgaga acgaggagga gttcaggatc gccctggagc aggtggccta ctacctgtcc 1500
gagaggaaga tgaacgagga gtccatcgag tacctgaaga agaacctggg cgccatcctg 1560
ctggagatct cctcctacga cctggagagg aacatcaccg gcgagtccaa ggagcacacc 1620
aggctgtggt ccgacttctg gaacccgaac aacaagaagg agtgcttctc caccaggctg 1680
aacccggagc tgaggatctt ctacaggccg ccgagggagc agaaggaccc gaagaagcag 1740
aagaacaggt tctccaagga ccacctggcc gtggccttca ccatcgccca gaacgccgcc 1800
aggaagagga tggagacctc cttcgccgag gagaaggacc tggtggagca ggtgaagaag 1860
ttcaacgagg aggtggtggg caagttcatc gacgagaagt ccgacaacct gtactactac 1920
ggcatcgaca ggggccagca ggagctggcc accctgtgcg tggtgaggtt ctccaaggag 1980
cactacgagg ccatgctgga ggacaacttc atcaagaagt tctccaagcc gatcccggcc 2040
cagatcaccg cctacaggat caaggacgag cacatgtcct acaggaagaa catcaccagg 2100
gacctgaagg gcaacgagac cgaggagatc ctgttcaaga acccgtccca cttcatcgac 2160
gaggtggaga acttcgagga ggtgtccacc ccgtgcatcg acctgaccac cgccaagctg 2220
atcaagggca agatcatcct gaacggcgac atccagacct acctggccct gaagaaggcc 2280
aacggcaaga ggcagctgtt cgagaagttc gccaagatcg acgactccgc caagatcgag 2340
ttcgacgact ccgagggcag gttccaggtg aagtccaagg ccaccgagag ggaggagtac 2400
cagttcctgc cgtactacgg cccggagcag gagaacatct ccccgaggga ggacatgagg 2460
agggagctgc aggcctacct ggacaagctg aggtcctccg agtccttcga ggaggacatc 2520
tccatcgaga agatcaacca cctgagggac gccatcacct ccaacatggt gggcatcatc 2580
gccttcctgt tcaccgagta cccgggcatc atcaacctgg agaacctgca ctccagggag 2640
aacatcgaga agaactggag gaagaacaac gaggacatct ccaggaggct ggagtggggc 2700
ctgtacaaga agttccagaa gatcggcctg gtgccgccga ggctgaggca gaccgtgctg 2760
ctgagggaga acgagaccga gaggcaggag aagctgaacc agttcggcat catccacttc 2820
atcccgaccg agaagacctc cgccaggtgc ccgtactgcg gcgagaacac cccgatgaag 2880
cagaggaacg aggacaagtt caagctgcac gcctacatct gcaggtccaa cgaggagaac 2940
tgcggcttcg acaccaggga gccgaagtcc ccgctggagt tcatcaagaa ctccgacgac 3000
gtggccgcct acaacatcgc caagaagagg ctgtccaggg ccgacccgaa gaagaagagg 3060
aaggtgtga 3069
<210> 102
<211> 3237
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 81所示融合蛋白的编码核酸序列
<400> 102
atgaagaacg gcatcaacct gttcaagacc aagaccacca agaccaaggg cgtggacatg 60
gagaagtacc agatcaccaa gaccatcagg ttcaagctgc tgccggacaa cgcccacgag 120
atcgtggaga aggtgaagtc cctgaagacc tccaacgtgg acgagctgat ggacgaggtg 180
aagaacgtgc acctgaaggg cctggagctg ctgttcgccc tgaagaagta cttctacttc 240
gacggcaacc agtgcaagtc cttcaagtcc accctggaga tcaaggccag gtggctgagg 300
ctgtacaccc cggaccagta ctacctgaag aagtcctcca agaactccta ccagctgaag 360
tccctgtcct acttcaagga cgtgttcaac gactggctgt tcaactggga ggagtccgtg 420
tccgagctgg ccatcatcta cgagaagtac aagatctgcc agcaccagag ggactccagg 480
gccgacatcg ccctgctgat caagaagctg tccatgaagg agtacttccc gttcatctcc 540
gacctgatcg actgcgtgaa cgacaagaac tccaacaaga ccttcctgat gaagctgtcc 600
gaggagctgt ccgtgctgct ggagaagtgc aactccaggg ccctgccgta ccagtccaac 660
ggcatcgtgg tgggcaaggc ctccctgaac tactacaccg tgtccaagtc cgagaagatg 720
ctgcagaacg agtacgagga cgtgtgccag tccctggaca agaactacga catcaccgag 780
atgaaggtga tcctgtacaa ggagaagctg gacaacctga acttcaagga cgtgaccatc 840
gccaacgcct acaacctgct gaaggagaac aaggccctgc agaagaggct gttctccgag 900
tacgtgtccc agggcaaggt gctgtccctg atcaagaccg agctgccgct gttctccaac 960
atcaacgaca acgacttcga gaagtacaag gagtggtcca acgagatcaa gaagctggcc 1020
gacaagaaga acaccttctg caagaagacc cagcaggaca agatcaagga catccagaac 1080
aagatctccg agctgaagaa gaagaggggc gccctgttcc agtacaagtt cacctccttc 1140
cagaagcact gcgacaacta caagaaggtg gccgtgcagt acggcaagct gaaggccagg 1200
aagaaggcca tcgagaagga cgagatcgag gccaacctgc tgaggtactg gtccgtgatc 1260
ctggagcagg aggacaagca ctccctggtg ctgatcccga agaacaacgc caaggacgcc 1320
aagcagtaca tcgagaccat caacaccaag ggcggcaagt acatcatcca ccacctggac 1380
tccctgaccc tgagggccct gaacaagctg tgcttcaacg ccgtggacat cgagaagggc 1440
cagatggtga gggagaacac cttctaccag ggcatcaagg aggagttcga gaggaacaag 1500
atcaactgcg acaaccaggg cgtgctgaag atccagggcc tgtactcctt caagaccgag 1560
ggcggccaga tcaacgagaa ggaggccgtg gagttcttca aggaggtgct gaagtccaac 1620
tacgccaggg aggtgctgaa cctgccgtac gacctggagt ccaacatctt ccagaaggag 1680
tacaccaacc tggaccagtt caggcaggac ctggagaagt gctgctacgc cctgcactcc 1740
aagatcggca aggacgacct ggacgagttc accaggaggt tcgaggccca ggtgttcgac 1800
atcacctcca tcgacctgaa gtccaagaag gagaagacca agaccaccgg cgagatgaag 1860
aagcacaccc agctgtggct ggagttctgg aagggcgcca tcgagcagaa cttcgccacc 1920
agggtgaacc cggagctgtc catcttctgg agggccccga agtcctccag ggagaagaag 1980
tacggcaagg gctccgacct gtacgacccg aacaagaaca acaggtacct gtacgagcag 2040
tacaccctgg ccctgaccat caccgagaac gccggctccc acttcaagga catcgccttc 2100
aaggacacct ccaagatcaa ggaggccatc aaggagttca acatgtccct gtcccagtcc 2160
aagtactgct tcggcatcga caggggcaac gccgagctgg tgtccctgtg cctgatcaag 2220
aacgagaagg acttcccgtt cgagaagttc ccggtgtaca ggctgaggga cctgacctac 2280
cagggcgact tcaaggacaa gcacgaccag atgaggtacg gcgtggccat caagaacatc 2340
tcctacttca tcgaccagga ggacctgttc gagaagaaca acctgtccgc catcgacatg 2400
accaccgcca agctgatcaa gaacaagatc gtgctgaacg gcgacgtgct gacctacctg 2460
aagctgaagg aggagaccgc caagcacaag ctgacccagt tcttccaggg ctcctccatc 2520
aacaagaact ccagggtgta cttcgacgag gacgagaacg tgttcaagat caccaccaac 2580
aggaaccaca acccggagga gatcatctac ttctacaggg gcgagtacgg cgccatcaag 2640
aacaagaacg acctggagga catcctgaac gagtacctgt gcaagatgga gaccggcgag 2700
tccgagatcg tgctgctgaa cagggtgaac cacctgaggg acgccatctc cgccaacatc 2760
gtgggcatcc tgtcctacct gatcgacctg ttcccggaga ccatcgtggc cctggagaac 2820
ctggccaagg gcaccatcga caggcacgtg tcccagtcct acgagaacat caccaggagg 2880
ttcgagtggg ccctgtacag gaagctgctg aacaagcagc tggccccgcc ggagctgaag 2940
gagaacatcc tgctgaggga gggcgacgac aagatcgacc agttcggcat catccacttc 3000
gtggaggaga agaacacctc caaggactgc ccgaactgca ggaagaccac ccagcagacc 3060
aacgacaaca agttcaagga gaagaagttc gtgtgcaagt cctgcggctt cgacacctcc 3120
aaggacagga agggcatgga ctccctgaac tccccggaca ccgtggccgc ctacaacgtg 3180
gccaggaaga agttcgagtc ctccagggcc gacccgaaga agaagaggaa ggtgtga 3237
<210> 103
<211> 3309
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 82所示融合蛋白的编码核酸序列
<400> 103
atggccggca ccccgtacac cggccacgtg gcctgcaagt actgcaagat cacctcctgg 60
gccacctacg acaggatcaa gatcaacaag atcaacatga accagtcctt catcaacggc 120
cagaacttct acgagctgag gaagaccatc aggttcgtgc tggacccgaa gaccctgaag 180
aggccgtaca ccccgtcctc cgacgaggtg aacctggagg agcagctgaa caacttcatc 240
gagaagtacc agcagggcat caacgacttc aagtacatcg tgtacttcgg cccgaagacc 300
gccgagacca aggagctgaa caagaagatc tccatcaagc actcctggct gaggaactac 360
accaagtccg agttctactc catcaaggac aagctgatcc agctggacta caacggcaac 420
aaggcctcca tcggcaactc caacctgaag ttcctgaacg agtacttcga gaactggatc 480
tccgagaacc aggagtgcgc cgacgccctg aagaactgca tcaacgcccc ggccgagaag 540
cagaagagga agtccgaggc cgcccactgg gtgaggaagc tgaccaagag gtccaacttc 600
gagtgcatct tcgagctgtt caacggcaac atcgaccaca agaactccaa cgacgacatc 660
gagaagatca agcactgcct gaacgagtgc aagaccctgc tgacctccct ggagaagatg 720
ctgctgccgt cccagtccct gggcatggag atcgagaggg cctccctgaa ctactacacc 780
atcaacaaga agccgaagaa ctacgacgag gacatcgccc agaaggcctc cgccctgaac 840
gaggcctacc agttcaaggc cgacgacaag gccttcctga acagggtggg cttctccgac 900
gacggcgtgc cgatcaacga gctgaaggag gccatgaaga agttcaaggc cgaccagaag 960
tccaagttct acgagttcgt gaaccagaag aagtcctact ccgacctgaa gaagaacgac 1020
gacctgaagc tgctgaacga catctccgag gaggacttca acaagttcaa ggagacccag 1080
gacaagatga ccaggggcaa gcacttccag ttctccttcc cgaactacaa gaagtccgag 1140
aagaacttct gcgacctgta caagaacgtg gccgtggcct tcggcaagat cagggccgac 1200
atcaaggccc tggagaagga gaggatggac gccgagaagc tgcagtgctg ggccgtgatc 1260
ctggagaagg acaaccagag gtacgtggtg accatcccga gggacgccaa caacaacctg 1320
accaacacca agcagtacat cgacaacctg cagaacgagg agaacgacca gtggatcctg 1380
tacgccttcg agtccctgac cctgaggtcc ctggacaagc tgtgcttcgg cctggacaag 1440
aacaccttca tcccggccat caccggcgag ctgtaccaga agaacaactc cttcttcgag 1500
aagggcctgc tgaagaggaa ggaccagttc tcccagaacg gcaccgacct ggccgccttc 1560
tacaagaccg tgctggagct ggactccacc aagaagatgc tgggcatcaa caagtacgcc 1620
gacttcaagg ccttcatctc caaggagtac accgccctgg aggacttcga gaagaccctg 1680
aaggagacct gctacttcaa gaagagggtg ttcatctccg aggacaccaa gaacaagctg 1740
atcaacgact accagggcaa cctgtacaag atcacctcct acgacctgga gaaggacgac 1800
tccgaggccc tgggcaccct gatcaacaag aagcagttca acagggcctc cccggagatc 1860
cacaccaaga cctggctgga cttctggacc gccgacaacg agaccgacaa gtacccgatc 1920
aggctgaacc cggagttcaa gatctccttc gtggagaagc aggacaagga cctgaacatg 1980
aggaacctgg gcctgctgaa caagaacagg aggctgaagt cccagttcct gctgtccacc 2040
accatcaccc tgctggccca cgagaagaac gccgacctgc acttcaagaa gaccgacgag 2100
atccagacct tcatcaactc ctacaaccag gagttcaaca agaagatcaa gccgttcgac 2160
atctactact acggcctgga caggggccag aaggagctgc tgaccctggg cctgttcaag 2220
ttctccgaga acgagaaggt gtccttcacc aagcaggacg gcaccgtggg cgagtactcc 2280
aagccgaagt tcatcccgct ggacgtgtac cagatcaggg agggccagta cctgaccaag 2340
aacaagaagg gcaggctggc ctacaagtcc atcgaccagt tcatcgacga cgagaaggtg 2400
atcgagaagc tgccggtgaa ctcctgcctg gacctgtcct gcgccaagct ggtgaagggc 2460
aagatcatcc agaacggcga cgtggccacc tacctggagc tgaagagggt gtccgccctg 2520
aggaagatct acgagaacac caccaggggc cagttcaaga ccgacaggat cggcttcaac 2580
aaggacaagg gctgcctgtt cctggacatc gagaacaggg gcaagctgga gaacaacaac 2640
ctgtacttct acgacaacag gttcgccgag atcctgtccc tggactccat catcaaggag 2700
ctgcaggact actacaacga ggtgaagaac aagcagaaca tcgagttcat ctccatcgac 2760
aagatcaacc acctgaggga cgccctgtgc gccaacgccg tgggcatcct ggcccacctg 2820
cagaagaccc acttcggcgt gatcgtgttc gagggcctgg acgccaggca caagaacaag 2880
gagaccaccg agttcgccgg caacctggcc tccaggatcg agaggaagat cctgcagaag 2940
ctggagaccc tgtccctgat cccgccgcag cacaggcaga tcatcgacct gcagaactcc 3000
aagcagatca agcagaccgg cgccgtgctg tacatcgagg agaagggcac ctccgccaac 3060
tgcccgcact gcgagaccgc caacccggac aagtccgaga agtggctggc ccacaactac 3120
aagtgcaaga actccaactg caacttcgac gcctccgaga tctccaagag gaaggacctg 3180
atcggcctgg acaactccga ctccgtggcc acctacaaca tcgccaagag gggcctgctg 3240
gagatgaacc agaagatcga gcagtccaag gtgtccaggg ccgacccgaa gaagaagagg 3300
aaggtgtga 3309
<210> 104
<211> 3207
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 83所示融合蛋白的编码核酸序列
<400> 104
atggagatcc aggagctgaa gaacctgtac gaggtgaaga agaccgtgag gttcgagctg 60
aagccgtcca agaagaagat cttcgagggc ggcgacgtga tcaagctgca gaaggacttc 120
gagaaggtgc agaagttctt cctggacatc ttcgtgtaca agaacgagca caccaagctg 180
gagttcaaga agaagaggga gatcaagtac acctggctga ggaccaacac caagaacgag 240
ttctacaact ggaggggcaa gtccgacacc ggcaagaact acgccctgaa caagatcggc 300
ttcctggccg aggagatcct gaggtggctg aacgagtggc aggagctgac caagtccctg 360
aaggacctga cccagaggga ggagcacaag caggagagga agtccgacat cgccttcgtg 420
ctgaggaact tcctgaagag gcagaacctg ccgttcatca aggacttctt caacgccgtg 480
atcgacatcc agggcaagca gggcaaggag tccgacgaca agatcaggaa gttcagggag 540
gagatcaagg agatcgagaa gaacctgaac gcctgctcca gggagtacct gccgacccag 600
tccaacggcg tgctgctgta caaggcctcc ttctcctact acaccctgaa caagaccccg 660
aaggagtacg aggacctgaa gaaggagaag gagtccgagc tgtcctccgt gctgctgaag 720
gagatctaca ggaggaagag gttcaacagg accaccaacc agaaggacac cctgttcgag 780
tgcacctccg actggctggt gaagatcaag ctgggcaagg acatctacga gtggaccctg 840
gacgaggcct accagaagat gaagatctgg aaggccaacc agaagtccaa cttcatcgag 900
gccgtggccg gcgacaagct gacccaccag aacttcagga agcagttccc gctgttcgac 960
gcctccgacg aggacttcga gaccttctac aggctgacca aggccctgga caagaacccg 1020
gagaacgcca agaagatcgc ccagaagagg ggcaagttct tcaacgcccc gaacgagacc 1080
gtgcagacca agaactacca cgagctgtgc gagctgtaca agaggatcgc cgtgaagagg 1140
ggcaagatca tcgccgagat caagggcatc gagaacgagg aggtgcagtc ccagctgctg 1200
acccactggg ccgtgatcgc cgaggagagg gacaagaagt tcatcgtgct gatcccgagg 1260
aagaacggcg gcaagctgga gaaccacaag aacgcccacg ccttcctgca ggagaaggac 1320
aggaaggagc cgaacgacat caaggtgtac cacttcaagt ccctgaccct gaggtccctg 1380
gagaagctgt gcttcaagga ggccaagaac accttcgccc cggagatcaa gaaggagacc 1440
aacccgaaga tctggttccc gacctacaag caggagtgga actccacccc ggagaggctg 1500
atcaagttct acaagcaggt gctgcagtcc aactacgccc agacctacct ggacctggtg 1560
gacttcggca acctgaacac cttcctggag acccacttca ccaccctgga ggagttcgag 1620
tccgacctgg agaagacctg ctacaccaag gtgccggtgt acttcgccaa gaaggagctg 1680
gagaccttcg ccgacgagtt cgaggccgag gtgttcgaga tcaccaccag gtccatctcc 1740
accgagtcca agaggaagga gaacgcccac gccgagatct ggagggactt ctggtccagg 1800
gagaacgagg aggagaacca catcaccagg ctgaacccgg aggtgtccgt gctgtacagg 1860
gacgagatca aggagaagtc caacacctcc aggaagaaca ggaagtccaa cgccaacaac 1920
aggttctccg acccgaggtt caccctggcc accaccatca ccctgaacgc cgacaagaag 1980
aagtccaacc tggccttcaa gaccgtggag gacatcaaca tccacatcga caacttcaac 2040
aagaagttct ccaagaactt ctccggcgag tgggtgtacg gcatcgacag gggcctgaag 2100
gagctggcca ccctgaacgt ggtgaagttc tccgacgtga agaacgtgtt cggcgtgtcc 2160
cagccgaagg agttcgccaa gatcccgatc tacaagctga gggacgagaa ggccatcctg 2220
aaggacgaga acggcctgtc cctgaagaac gccaagggcg aggccaggaa ggtgatcgac 2280
aacatctccg acgtgctgga ggagggcaag gagccggact ccaccctgtt cgagaagagg 2340
gaggtgtcct ccatcgacct gaccagggcc aagctgatca agggccacat catctccaac 2400
ggcgaccaga agacctacct gaagctgaag gagacctccg ccaagaggag gatcttcgag 2460
ctgttctcca ccgccaagat cgacaagtcc tcccagttcc acgtgaggaa gaccatcgag 2520
ctgtccggca ccaagatcta ctggctgtgc gagtggcaga ggcaggactc ctggaggacc 2580
gagaaggtgt ccctgaggaa caccctgaag ggctacctgc agaacctgga cctgaagaac 2640
aggttcgaga acatcgagac catcgagaag atcaaccacc tgagggacgc catcaccgcc 2700
aacatggtgg gcatcctgtc ccacctgcag aacaagctgg agatgcaggg cgtgatcgcc 2760
ctggagaacc tggacaccgt gagggagcag tccaacaaga agatgatcga cgagcacttc 2820
gagcagtcca acgagcacgt gtccaggagg ctggagtggg ccctgtactg caagttcgcc 2880
aacaccggcg aggtgccgcc gcagatcaag gagtccatct tcctgaggga cgagttcaag 2940
gtgtgccaga tcggcatcct gaacttcatc gacgtgaagg gcacctcctc caactgcccg 3000
aactgcgacc aggagtccag gaagaccggc tcccacttca tctgcaactt ccagaacaac 3060
tgcatcttct cctccaagga gaacaggaac ctgctggagc agaacctgca caactccgac 3120
gacgtggccg ccttcaacat cgccaagagg ggcctggaga tcgtgaaggt gtccagggcc 3180
gacccgaaga agaagaggaa ggtgtga 3207
<210> 105
<211> 3510
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 84所示融合蛋白的编码核酸序列
<400> 105
atggagaact tcaagaacct gtacgaggtg aggaagaccg tgaggttcga gctgaagccg 60
tccaggaaga agaccttcgc cggcggcgac atcttcgagc tgcagaagga cttcgaggag 120
gtgcagaagt tcttcctgga catcttcgtg ttcgccatcg agcaggagaa gctgtaccag 180
gaggaggagg aggagggcaa gctgtccagg tacaccaaga tcgagttcaa gaagaagagg 240
gagatcaagt acacctggct gaggatctac accaagaacg agttctacga ctggaacggc 300
aagaacgaca aggagaagaa ctacgccctg tccaagatcg acttcctgga gaaggagatc 360
ctgaggtggt tcaacgagtg gcaggagctg accgtgaacc tgaagaacct gacccagacc 420
aaggagcacg agaaggagag gaagtccgac atcgccttcg tgctgaggaa cttcctgaag 480
aggcagaact tcccgttcat caaggacttc ttcaacgccg tgatcgacat ccaggagaag 540
cagggcaacg agtccgacga gaagatcagg aagttcaggg aggagctgag ggagatgaag 600
aagaacctga acacctgcgc caaggagtac ctgtcctccc agtccaaggg cgtgctgctg 660
cacaaggcct ccttcaacta ctacaccctg aacaagaccc cgaaggagta cgagaacctg 720
aagctgcaga aggagctgga gatcgacaac atcctgccga agaagatctg caagagggtg 780
aggtggaaca aggagaagaa gcaggaggac atcctgttcg agtgcaactc cgactggctg 840
gtggagatca agctgggcta cgacatccag aagtggaccc tggacgaggc ctaccagaag 900
atgaagacct ggaaggccga ccagaagtcc gacttcaacg agaagatcgg caacttcatc 960
gaccagtacc tgaagaaggg cttcatcgag gacctgatga acgagaacga gaagaagaac 1020
gccgaggcca tcctgaggga gttctccgtg ttcaagccga tcgagaactt ctacttctac 1080
gacttcctgg agaggaccaa ggagatcaag atcctgtcca accagaagaa caacatcctg 1140
cagaagtaca acaagaacgc caagtacttc gagaagatca tcacctacaa gatcaaggac 1200
aaggaggacc tgaccgagga cgagaaggag taccaggagc tggagaagtc catcgagaag 1260
aaggccaagg agaggggcaa gttcttcaac gccccgaagg agaaggtgca gacccagcac 1320
tacttcgagc tgtgcgagct gtacaagagg atcgccatga agaggggcaa gatcatcgcc 1380
gagatcaagg gcatcgagaa cgaggaggtg cagtcccagc tgctgaccca ctgggccctg 1440
atcgccgagg agggcgagaa gaagtccgtg gtgttcatcc cgaggaagaa cggcgaggag 1500
ctggagaacc acaagaaggc ccacgagttc ctgcagaagc aggagaagaa ggagttcggc 1560
gacatcaagt cctaccactt caagtccctg accctgaggg ccctggagaa gctgtgcttc 1620
aaggagaccg agaacacctt caccccggag atcaagaagg agaccaaccc gaaggtgtgg 1680
ttcccgaagt acaagcagga gtggaacgac gagccgcaga agctgatcaa cttctacaag 1740
caggtgctgc agtccaagta ctcccagaag tacctggacc tggtggcctt cggcgacctg 1800
aagtccttcc tggagacctc cttcgacgac ctgcagatct tcgagtccgg cctggagaag 1860
acctgctaca tcaaggtgcc gatctacttc tccaaggagg gcttcgagac cttcaccaac 1920
aggttcgacg ccgaggtgtt cgagatcacc accaggtcca tctcctccga gtccaagagg 1980
aaggagaacg cccacgccga gatctggaag gacttctggt ccaaggagaa cgaggagaag 2040
aaccacatca ccaggctgaa cccggaggtg tccgtgttct acagggacga gatcgagaag 2100
aagtccaacg ccctgagggg caacaacaag tccaacatca acaacaggtt ctccgcctcc 2160
aggttcaccc tggtgaccac catcaccatc agggccaccc acaagaagtc caacctggcc 2220
ttcaagaccg aggaggacat caagtcccac atcgacaagt tcaacgaggc cttccagaac 2280
ttctccggcg agtgggtgta cggcatcgac aggggcctga aggagctggc caccctgaac 2340
gtggtgaagt tctccgacga gaagaacgag ttcggcgtga tcaagccgaa ggagttcgcc 2400
aagatcccgg tgtacaagct gaaggacgag aaggccatcc tgaaggacga gaacggcaag 2460
gacctgaaga acgccaaggg cgaggccagg aaggtgatcg acaacatctc cgaggtgctg 2520
gaggagaaga aggagccgga ctccaacctg ttcgagaagc agggcgtgct gtcccagggc 2580
atctcctgca tcgacctgac ccaggccaag ctgatcaagg gccacatcat cctgaacggc 2640
gaccagaaga cctacctgaa gctgaaggag atctccgcca agaggaggat cttcgagctg 2700
ttctccacct ccaagatcga caagaactcc gagctgaggg tggagaagac caccatctcc 2760
atcaactccg aggacggcaa gagggacttc tactggctga ccaagaacca gatcgtgaac 2820
tccgagacca agaaggagat ccagaaggag cagcaggaga agctggacaa cctgaaggtg 2880
atcttcatcg actacctgga gggcctgtgc gtgaagaaca agttcgagga catcgagacc 2940
atcgagaaga tcaaccacct gagggacgcc atcaccgcca acatggtggg catcctgttc 3000
cacctgcaga aggagttcaa gggcatcatc gccctggaga acctggacac cgtgagggag 3060
cagtccaaca agaagatgat cgacgagcac ttcgagcagt ccaacgagga catctccagg 3120
aggctggagt gggccctgta caggaagttc gccaacatgg gcgaggtgcc gtcccagatc 3180
aaggagtcca tcttcctgag ggacgagttc aaggtgtacc agatgggcct gctgaagttc 3240
gtggaggtgt ccggcacctc ctccaactgc ccgaactgcg acaaggaggt gggcaagacc 3300
aactcccact tcgtgtgcaa gggcgagaac aactgcggct tctcctccaa ggagaacagg 3360
aacctgctgg agcagaacct gaacaactcc gacgaggtgg ccgcctacaa catcgccaag 3420
aggggcctga agctgatcaa ccagaagtgg aacaacacct ccaagtccca gaactccagg 3480
gccgacccga agaagaagag gaaggtgtga 3510
<210> 106
<211> 3228
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 85所示融合蛋白的编码核酸序列
<400> 106
atggagaagt acaagatcac caagaccatc aggttcaagc tgctgccgga caagatccag 60
gacatctcca ggcaggtggc cgtgctgcag aactccacca acgccgagaa gaagaacaac 120
ctgctgaggc tggtgcagag gggccaggag ctgccgaagc tgctgaacga gtacatcagg 180
tactccgaca accacaagct gaagtccaac gtgaccgtgc acttcaggtg gctgaggctg 240
ttcaccaagg acctgttcta caactggaag aaggacaaca ccgagaagaa gatcaagatc 300
tccgacgtgg tgtacctgtc ccacgtgttc gaggccttcc tgaaggagtg ggagtccacc 360
atcgagaggg tgaacgccga ctgcaacaag ccggaggagt ccaagaccag ggacgccgag 420
atcgccctgt ccatcaggaa gctgggcatc aagcaccagc tgccgttcat caagggcttc 480
gtggacaact ccaacgacaa gaactccgag gacaccaagt ccaagctgac cgccctgctg 540
tccgagttcg aggccgtgct gaagatctgc gagcagaact acctgccgtc ccagtcctcc 600
ggcatcgcca tcgccaaggc ctccttcaac tactacacca tcaacaagaa gcagaaggac 660
ttcgaggccg agatcgtggc cctgaagaag cagctgcacg ccaggtacgg caacaagaag 720
tacgaccagc tgctgaggga gctgaacctg atcccgctga aggagctgcc gctgaaggag 780
ctgccgctga tcgagttcta ctccgagatc aagaagagga agtccaccaa gaagtccgag 840
ttcctggagg ccgtgtccaa cggcctggtg ttcgacgacc tgaagtccaa gttcccgctg 900
ttccagaccg agtccaacaa gtacgacgag tacctgaagc tgtccaacaa gatcacccag 960
aagtccaccg ccaagtccct gctgtccaag gactccccgg aggcccagaa gctgcagacc 1020
gagatcacca agctgaagaa gaacaggggc gagtacttca agaaggcctt cggcaagtac 1080
gtgcagctgt gcgagctgta caaggagatc gccggcaaga ggggcaagct gaagggccag 1140
atcaagggca tcgagaacga gaggatcgac tcccagaggc tgcagtactg ggccctggtg 1200
ctggaggaca acctgaagca ctccctgatc ctgatcccga aggagaagac caacgagctg 1260
tacaggaagg tgtggggcgc caaggacgac ggcgcctcct cctcctcctc ctccaccctg 1320
tactacttcg agtccatgac ctacagggcc ctgaggaagc tgtgcttcgg catcaacggc 1380
aacaccttcc tgccggagat ccagaaggag ctgccgcagt acaaccagaa ggagttcggc 1440
gagttctgct tccacaagtc caacgacgac aaggagatcg acgagccgaa gctgatctcc 1500
ttctaccagt ccgtgctgaa gaccgacttc gtgaagaaca ccctggccct gccgcagtcc 1560
gtgttcaacg aggtggccat ccagtccttc gagaccaggc aggacttcca gatcgccctg 1620
gagaagtgct gctacgccaa gaagcagatc atctccgagt ccctgaagaa ggagatcctg 1680
gagaactaca acacccagat cttcaagatc acctccctgg acctgcagag gtccgagcag 1740
aagaacctga agggccacac caggatctgg aacaggttct ggaccaagca gaacgaggag 1800
atcaactaca acctgaggct gaacccggag atcgccatcg tgtggaggaa ggccaagaag 1860
accaggatcg agaagtacgg cgagaggtcc gtgctgtacg agccggagaa gaggaacagg 1920
tacctgcacg agcagtacac cctgtgcacc accgtgaccg acaacgccct gaacaacgag 1980
atcaccttcg ccttcgagga caccaagaag aagggcaccg agatcgtgaa gtacaacgag 2040
aagatcaacc agaccctgaa gaaggagttc aacaagaacc agctgtggtt ctacggcatc 2100
gacgccggcg agatcgagct ggccaccctg gccctgatga acaaggacaa ggagccgcag 2160
ctgttcaccg tgtacgagct gaagaagctg gacttcttca agcacggcta catctacaac 2220
aaggagaggg agctggtgat cagggagaag ccgtacaagg ccatccagaa cctgtcctac 2280
ttcctgaacg aggagctgta cgagaagacc ttcagggacg gcaagttcaa cgagacctac 2340
aacgagctgt tcaaggagaa gcacgtgtcc gccatcgacc tgaccaccgc caaggtgatc 2400
aacggcaaga tcatcctgaa cggcgacatg atcaccttcc tgaacctgag gatcctgcac 2460
gcccagagga agatctacga ggagctgatc gagaacccgc acgccgagct gaaggagaag 2520
gactacaagc tgtacttcga gatcgagggc aaggacaagg acatctacat ctccaggctg 2580
gacttcgagt acatcaagcc gtaccaggag atctccaact acctgttcgc ctacttcgcc 2640
tcccagcaga tcaacgaggc cagggaggag gagcagatca accagaccaa gagggccctg 2700
gccggcaaca tgatcggcgt gatctactac ctgtaccaga agtacagggg catcatctcc 2760
atcgaggacc tgaagcagac caaggtggag tccgacagga acaagttcga gggcaacatc 2820
gagaggccgc tggagtgggc cctgtacagg aagttccagc aggagggcta cgtgccgccg 2880
atctccgagc tgatcaagct gagggagctg gagaagttcc cgctgaagga cgtgaagcag 2940
ccgaagtacg agaacatcca gcagttcggc atcatcaagt tcgtgtcccc ggaggagacc 3000
tccaccacct gcccgaagtg cctgaggagg ttcaaggact acgacaagaa caagcaggag 3060
ggcttctgca agtgccagtg cggcttcgac accaggaacg acctgaaggg cttcgagggc 3120
ctgaacgacc cggacaaggt ggccgccttc aacatcgcca agaggggctt cgaggacctg 3180
cagaagtaca agtccagggc cgacccgaag aagaagagga aggtgtga 3228
<210> 107
<211> 3354
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 86所示融合蛋白的编码核酸序列
<400> 107
atgctgatcc agttcaagaa ccactactcc tacaacaagt ccatcaggtt caagctggag 60
cacaagaacg gcaagctgcc gaagctggag tccgacaacg tggacctgaa caagctggtg 120
gacatcggca actccctgaa ggacatcttc gaggagctgg tgtacaccaa gaacaactac 180
aacaagctga actccctggt gtccatcaag aagcagtggc tgaagatcta cttcaagaac 240
gagttctact ccaacggcaa gatccagaac tactccctgt ccaacttctc ctacctgccg 300
aacaagctga tcgagtggct gaacaactgg cagaacaacc tgaaggccct gatcgagctg 360
accaagcagc aggacttcaa caagaccaag aagtccgaga tcgcctacat cctgtccctg 420
ttcaacggca agtactcctt ctccttcgtg aaggacttct ccacctgcat caaccacaag 480
aactcccagg agcagatcct gaagctgcag ggcgtggtgg agaacttcga gaaggtgctg 540
aacctgtgca tccaggagta cctgccgtcc aagtccgccg gcgtggtgat cgcccagggc 600
tccatgaact actacgccat caacaaggag ccgaagaggt acgacaacat cctggccgac 660
ctgaaccaga agttcgagga gctggacaag gagtacatcg ccatgaagca gtacaagtcc 720
tcccagaagt ccaggctgtt cgagttcatc aggaagggct tctccaagga ccagatcctg 780
tccgagttca agaagaagga gaacaacgag gtgtccttcg tgtacaacaa ccagatcatc 840
atcaggatct acacccagga gctgttcaag gactcctact gcctgggcga ggtgatcaag 900
ctgaccaaga agatcgagga gctgaacgag tccaaggact ccaacaacaa cctgccggag 960
gagaccaaga aggagatcac caagctgaag aaggagatcg gcttctactt catcaggagg 1020
accaggggca agtcccacaa caactacttc aagtcctact acggcttctg caacgacaag 1080
ttcaagaaga aggcccagga gaggggcagg ctgctgacca agatcaaggc catcaggaag 1140
gagaagatcg agtcccagaa cctgaggtac tggtccctga tcctggacga cggcaaggac 1200
aagttcctgt ggctggtgcc gaaggagaac atgcaggagt tcaggaggga gctgtccaag 1260
atccacccgt ccggcgagtc ctccctgttc ctgttccact ccctgaccat gagggccctg 1320
cacaagctgt gcttcgccca ggagtccgac ttcgtgaagg agatgccgaa ggtgctgaag 1380
gaggagcagc tgaactgcga gaaggcctcc aacgacaccg agaccaacaa gaggatcaag 1440
aggaacttcg gcctgaacta catcaagacc aaggacgagc tgaccctgtc cttcctgaag 1500
aagctgatca tctccgagta cgcccacgag aggctggacc tgaaccactt cgacctgtcc 1560
aagctgcagg tggccaccac cctgaacgag ttcgaggagt acctggagga cgcctgctac 1620
tacctggaga agatctccat ctcctcctcc atgatcaagg agctgctgga ggagtacaac 1680
atcctgaact tcaggatcac ctcctacgac ctggagaaga ggaacaagaa cacctaccag 1740
accccggagt ccgacatcaa gaggcacacc aaggagatct ggaacaagtt ctgggagggc 1800
gacaggttca tcaggctgaa cccggagatc aagatcaggt acaggcagaa gaaccagaac 1860
atcgaggact acctgaagga gaagggcttc gacctgacca agatcaagaa caggttcctg 1920
caggagcagt actccgtgtc cttcaccttc gccctgaacg ccggcaagaa gtacccgaag 1980
ctggccttcg tgaagaccga ggagatcctg gagaagatcg aggagttcaa cgacgagttc 2040
aacaagcagt acttcgacaa ctcctacaag tacggcatcg acaggggcaa catcgagctg 2100
gccaccctgt gcatcaccaa gttcaacaag aacgacacct acgagtacaa gggcaagaag 2160
tacctgaagc cgaacttccc gacctcccag gaggacatca agacctacga gctgaagaac 2220
gagtggtaca agaggaccgc catctccaac atcgagacca agccgaagaa caagaagacc 2280
ccgaagagga tcatcgccaa catctcctac ttcatcgaca acgtggagaa cgaggagtgg 2340
ttcaacaaga agacctgcac ctccatcgac ctgaccaccg ccaaggtgat caagggcaag 2400
ctgatcctga acggcgacgt gctgaccttc ctgaagctga agaaggaggc cgccaagagg 2460
atcctgttcg agctggtggc ccagaacaag ctgaccgcca agaacaagga gctgaagtgg 2520
aagtccgacg acggcaacaa ctccgactcc gtgaggctga tctgcgacgt gctggacaac 2580
gagaccaact ccatctactt ctacgaggac tccaagtacg gcaggggctt cgagggcctg 2640
ctgaccaccg acaagaccgc ctactccaag gagggcatca ggatcaacct gcagaactac 2700
ctgaaccacc tgatctccga gaaggagaac aagtccaaca aggcctactc ccacgtgccg 2760
tccatcgaga agatcaacca cctgagggac gccctggtgg ccaacatggt gggcgtgatc 2820
tcctacctgc aggcctacta cccgggcatc gtggtgctgg aggacctgaa ccacaagctg 2880
ctgatcaagc acttcgagga cctgaacatc aacatctcca acaggttcga gcacgccctg 2940
atcgagaagt tccagaccct gggcatggtg ccgccgcaca tcaaggacta cctggagatc 3000
aggtcctcct tcaggatgtc caggaacgac tcctcccagt tcggcgccct gatcttcgtg 3060
tccaaggagg gcacctccaa ggagtgcccg tactgcgaga agaagtggaa ctggggcaag 3120
gagaaggaga tcgagctgaa gttctccaag aagcagtaca tctgcggcaa ggagaactcc 3180
tgcggcttcg acaccaagca catccagaac accttcgagt tcctgtccga gatcaacgac 3240
ccggacaaga tcgccgccta caacatcgcc aagaggggct tcaagtcctt catcaacaag 3300
tcctccatca agaagcagtc cagggccgac ccgaagaaga agaggaaggt gtga 3354
<210> 108
<211> 3291
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 87所示融合蛋白的编码核酸序列
<400> 108
atggagaagt acaagatcac caagaccatc aggttcaagc tgctgccgga caagatccag 60
gacatctcca ggcaggtggc cgtgctgcag aactccacca acgccgagaa gaagaacaac 120
ctgctgaggc tgatccagag gggccaggag ctgccgaagc tgctgaacga gtacatcagg 180
tactccgaca accacaagct gaagtccaac gtgaccgtgc acttcaggtg gctgaggctg 240
ttcaccaagg acctgttcta caactggaag aaggacaaca ccgagaagaa gatcaagatc 300
tccgacgtgg actacctgtc cagggtgttc gaggacttct tcaacgagtg ggagaccgtg 360
atcgagagga tcaacaccga ctgcaacagg ccggaggagt ccaagaccag ggacgccgag 420
atcgccttct ccatcaagaa gatcgccacc aagcagatgt tcccgttcat caagtccttc 480
gtgtacaact ccaactacaa gaactccgag gagaccaagt ccaagctgac cgccctgctg 540
aacgagttcg agaccgtgct gaagatctgc gagcagaact acctgccgtc ccagtccgcc 600
ggcatcgtga tcgccaaggc ctccttcaac tactacacca tcaacaagaa gcagaaggac 660
tacaagggct acaccgacga catcgagaag atcgagaagg gcatgaactc caagttccac 720
tacgagagga agtacgacca gctgctggag gagctgaacc tgatcgccct gaaggagctg 780
ccgctgatcg agttctactc caagatcaag tcctacaagt ccaccaggaa gatcgagttc 840
tccgaggccg tgtccaaggg cctggccttc gccgacctga agtccaagtt cccgctgttc 900
cagaccgagt ccaacaagta cgccgagttc ctggagctga ccggcaggat cacccagatc 960
tccaccgcca agtccctgct gtccaaggac aacccggagg cccagaagct gagggacgag 1020
atcaagaagc tgaggatcaa caggggcgag tacttcaaga acaacttcca caagtacatc 1080
tccctgtgca acctgtacaa gaagatcgcc gacaagaagg gcaggctgaa gggccaggtg 1140
aagggcatcg agaacgagag gatcgactcc cagaggatcc agcactgggc cctggtgctg 1200
gaggacaacc tgaagcactc cctgatcctg atcccgaagg agaaggtgac cgaggtgtac 1260
aggaaggtga gggcctccaa ggccgactcc acctcctcct cctcctccct gtactacttc 1320
gagtccatga cctacagggc cctgcacaag ctgtgcttcg gcgtgaacgg caacaccttc 1380
ctgccggaga tccagaagga gctgccggag tacaacccga acaagcagtc cgacttcggc 1440
gagttctgct tccacaagtc caacaccgac aaggagatcg acgagccgaa gctgatctcc 1500
ttctaccagt ccgtgctgaa gaccaactac gtgaaggaca acctgaacct gccgcagtcc 1560
gtgttcgacg aggccaccgt gcagaccttc gagaccaggc aggacttcca gatcgccctg 1620
gagaagtgct gctacgccaa gaagaccatc atctccgaga ccctgaagaa ggagatcctg 1680
gaggacaaca acgtgcagat cttccagatc acctccctgg acctgcagag gtccgagcag 1740
aagaacctga aggcccacac caagatctgg aacaggttct ggaccaagca gaacgagacc 1800
gccaactacg acctgaggct gaacccggag accgccatcg tgtggaggaa gccgaagaag 1860
accaggatcg acaagtacgg cgccggcacc tccctgtacg acccgaagaa gaggaacagg 1920
tacctgcacg agcagtacac cctgtgcacc accgtgaccg acaacgccct gaacaacgag 1980
atcaccttcg ccttcgagga caccaagaag aagggcaccg agatcgtgaa gtacaacgag 2040
aagatcaacc agaccctgaa gaaggagttc aacaagaacc agctgtggtt ctacggcatc 2100
gacgccggcg agatcgagct ggccaccctg gccctgatga acaaggacaa ggagccgcag 2160
ctgttcaccg tgtacgagct gaagaagtcc gacttcttca agcacggcta catctacaac 2220
aaggagaggg agctggtgat cagggagaag ccgtacaagg ccatccagaa cctgtcctac 2280
ttcctgaacg aggagctgta cgagaagacc ttcagggacg gcaagttcca ggagaccttc 2340
aacgagctgt tcaaggagaa gcacgtgtcc gccatcgacc tgaccaccgc caaggtgatc 2400
aacggcaaga tcatcctgaa cggcgacatg atcaccttcc tgaacctgag gatcctgcac 2460
gccaagagga agatctacga ggagctgatc atcaacccgc aggccgagct gaaggagaac 2520
gagaaggagt actacctgta cttcgacaag gagggcaccg agaaggtgga gaagatctac 2580
aggtccaggc tggacttcga gcacatcaag ccgtaccagg agatcaggaa cgacctgaac 2640
gcctacttca agaacgtgca gaagaacgag gccaaggtgg aggaccagat caaccagacc 2700
aggagggccc tggtgggcaa catgatcggc gtgatctact acctgtacca gaagtacagg 2760
ggcatcatct ccatcgagga cctgaagcag accaaggtgg agtccgacag gaacaagttc 2820
gagggcaaca tcgagaggcc gctggagtgg gccctgtaca ggaagttcca gcaggagggc 2880
tacgtgccgc cgatctccga gctgatcaag ctgagggagc tggagaagtt cccgctgaag 2940
gacgtgaagc agccgaagta cgagaacatc cagcagttcg gcatcatcaa gttcgtgtcc 3000
ccggaggaga cctccaccac ctgcccgtcc tgcgagaaga aggcctacga gctgcagaag 3060
gagaagaagg gcgaggagaa gccggccgag aacaagaggt acgaggccga caagaaggcc 3120
ggcgtgttct gctgcccgaa gtgcggcttc cacaacagga ccaacccgat gggctacgag 3180
tccctggact ccaacgacaa ggtggccgcc ttcaacatcg ccaagagggg cttcgaggac 3240
ctgcagaagc acaagtccag ggccgacccg aagaagaaga ggaaggtgtg a 3291
<210> 109
<211> 3135
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 88所示融合蛋白的编码核酸序列
<400> 109
atggagaact ccaacctgta ccaggtggtg aagaccatca ggttcaagct ggagccggtg 60
ggcaagatgg acaccccgaa gttcggcgac aagaacgccg agtccaaggc caacctgacc 120
ccgttcatcg agctggtgaa gaagaccatg accaacgtga aggccctggt gttctccaag 180
caggacggcg aggacggcga gaagtggagg aagatcctgg aggtgaacta caggttcctg 240
aggtcctacc tgaagaactc cttctacgag aacaggggcg actcccagga gaagtccaag 300
aagcacaaga tctccgacct ggagtacctg cagaaggccc tggagaacct gttcgccgag 360
ttcgacgaga tcctggacgg cctggaggac ttcgagaaga ggaacaccaa gaaccagtac 420
gagaagcaga ggcacgccca ggccggcctg ctgctgaaca ggctgtgcaa gaggtccaac 480
ttcggcttcc tgaaggcctt cgtgggcgcc ctggcccaga ccaacaagcc gttcttcgac 540
gacaagaccg acaagctgaa gaagcagatc gacaagttcg agaccgagct ggagaagcag 600
aaggagttct tcctgccgta ccagtccaac ggcgtgctgt tcgccggcgg ctccttcaac 660
aggtacgcca tcaacaagac cccgaagatg ctggacaagg agctgaggga ggagcagacc 720
aacctgaaga agtccctgtg cgagcacaag atcaagatcg acaccctgaa caccctgggc 780
ctgaagaacg actgcccgtg cacctccctg gacaactcct acaccttcat caaggactac 840
aaggccaagc agaagtccaa gttcatcgag ctggtgcaga agggcgagtt cgacgaggcc 900
aagaaggtga acctgttcga gtgctccgag accgacttcg agaccttcaa gaccaggacc 960
aagcagatcc agaacgagaa ggacaaggac gagaggacca agctgaagca gaagaggggc 1020
gagttcttca agtcccagaa gaggggcaag ttcttcaagt cccagaccca gaactacgag 1080
aacctgtgcg acctgtacaa gaagatcgcc cagaagaggg gccagatcgt ggccaagatc 1140
tgcgccatca agaaggagaa ggagatgtgc gagcaggtga agtactggtg cgtggccctg 1200
gagaagggcg gcgagttcta cctgtacatg ttcctgaggg acgagaacga caacatcaag 1260
aacgcctacg acttcgtgtc caagctgcag acccagaagt ccggcgagac caagctgcac 1320
tacttcgact ccctgaccct gaaggccgtg aggaagctgt gcttcaagga gaccgacggc 1380
tccttcaaga aggccctgaa gaacgtgaag ttcccggagt gcgagcagaa cctggacgag 1440
aaggtgaaga tctccttcta ccagaacgtg ctgaagaacg ccaagaccct gaacctgtcc 1500
aagttcgaga acctgcagtc cgtgaccgag ggcaagttcg agtccctgtc cgagttcgag 1560
gtggccctga acatggtgtg ctacaccaag accgtgtgcg tgtccgagtc cgtggagaag 1620
gagctgaaga agttcaagcc gctggtgttc cacatcacct cccaggacct ggccgccaag 1680
agggagaaga aggcccacac ccagatctgg cacgagttct ggagggagtc caacgagaag 1740
tccaagttcc cgctgaggct gaacccggag ctgaaggtga tgtggaggga ggccaggccg 1800
tccagggtgg agaagtacgc cgagcagtcc gacaagttcg acccgaacaa gaagaacagg 1860
tacctgcacc cgcagttcac cctggccctg aacttcaccc agaacgccca caacgaggcc 1920
atcaacctgg ccttcaagga cgtgcagaac aagggcgagg ccgtgaagaa gttcaacgag 1980
aacttcaagt cctccgagta cgccttcggc atcgacgtgg gcaccaagga cctggccctg 2040
ctgtgcctga tcgacaagaa caagaagccg gtgaacttcg acgtgtacga gatctgcaac 2100
gagaacgaga tctgcaacga gaagctgggc ttcgagaagt tcggcttcta caaggacggc 2160
accaggaggg acgagccgta caagctgatc aagaacccgt cctacttcct gaacgagtcc 2220
ctgtacaaga agaccttcaa cgccaccaag gaggagttcg agaggtcctt ctccgagctg 2280
ttcaagagga agtccgtgtg cgccctggac ctgaccaccg ccaaggtgat ctgcggcaag 2340
atcatcctga acggcgactt ctccacccac ctgaacctga agatcctgaa cgccaagagg 2400
aagatctccg ccaagctgaa gaaggacccg accctgaaga tcgagtacga caacgacgac 2460
aacatcctgt tcggctccaa cgtgatcttc tactacaaca acaagtacga gatcgtgagg 2520
ccgtacgacg agatcaagaa cgagatcttc gagttccacg agaagcagag gctggacgac 2580
gccaggctgg aggacaacat caacaagacc agggccaacc tggtggccaa catggtgggc 2640
gtgatctcct tcctgcacaa ggagttctcc ggcttcgtgg tgctggagaa cctgaagcag 2700
tccgagatcg agggcaacca caggctgaag ttcgagggcg acatcaccag gccgctggag 2760
ctggccctgt acaggaagtt ccagtccaag tgcctgaccc cgccgatctc cgagctgatc 2820
aagctgaggg agggcgagaa gaacgagaac gtggagtccg acctgatcct gcagttcggc 2880
atcatcaagt tcgtggacaa ggacaagacc tccaggctgt gcccggcctg cggcaaggac 2940
gcctacgaga acaacaactc caagtacaag accgacaaga aggacggcgt gttcgagtgc 3000
gccggctgcg gcttcaacaa caagaacaac gccggcgact tcgccgccct ggacaccaac 3060
gacaagatcg ccaccttcaa catcgccaag aggggcctgt ccagggccga cccgaagaag 3120
aagaggaagg tgtga 3135
<210> 110
<211> 3540
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 89所示融合蛋白的编码核酸序列
<400> 110
atggagacct acaagatcac caagaccatc aggttcaagc tggaggccga cgaggagaac 60
tccatccaca tcaaggagga catcatcaac atcgagacca acgacaacga gttcaccatg 120
gtggacttcg tgtccaacct gggcaactac atcaaggacc tgaagaacta cctgttctac 180
gagaagaagg acggctccct gtccttcaag gacaagatca tcatcaagaa cgagtggctg 240
aggcagtacg ccaagcagga cttcgtggag ctgaagtcca agaagaggat caacctgagg 300
aacaacagga tggagcagat caagatcggc gacatcccga ggctgtcctc caagatcgag 360
gaggccctgg acatcgccaa ggagatctac tccaagctgt ccgacgacgc caccctggag 420
cagcacgaga ggaccaagaa ggcccagatc ggcctgctgc tgaagaggct ggaggccaag 480
aacgtgctgc cgctgctgat ggacctggtg aaggagaccc tggacaagga cgagaccgac 540
gacctgtcca tcaggctgaa gaggcagtcc cagaagatca actcccagct gaagatcgcc 600
atcaggtcct tcctgccgga gcagtccaac ggcctgcaga tcgccaaggc ctccttcaac 660
tactacacca tcaacaagaa gccgatcgac ttcgagaaga agatcgagga cctgaagaag 720
aacctgaacg tgaaggacct ggagaagctg aacgtgtact tcgacaagaa ggagaagaag 780
cagaagaact acctgggcaa gaagatcttc tccctgttcg agaccgacat ccagaaggcc 840
ctgtccaaga accagccgct gtacctgggc gacgccccga tgatcgactc cgcctacgtg 900
tccctgaggc agatcttcaa gaagatcaag tccgagcaga agaagcagtt ctccgagctg 960
atgcagaaca agtgctccta cgacgagctg aagaactcca acctgtacct gctgaacgac 1020
atcggcctgg agcagttcaa cacctacagg gagaagacca aggagctgga ggagctggcc 1080
accaagctgt ccaaccagaa cctgctggag aacgccaagg agaggctgag gtcccagaag 1140
gagaagatcg ccaaggagag gggcaacatc atgaaggaca ggttccagac ctggaagtcc 1200
ttcgccaact tctacaggac cgtgtcccag aagcacggca agatcctggc ccagctgaag 1260
ggcatcgaga aggagcaggc cgagtcccag ctgctgaagt actgggccct gatctgcgag 1320
aaggagaacc agcaccagct gtggctgatc ccgagggaga aggcctggga gtgcaagagg 1380
tggctggaga ccgtgaacga cacctccatc gacaacgaga actccatcaa gctgtactgg 1440
ttcgagtccc tgacctacag gtccctgcag aagctgtgct tcggcttcct ggagaacggc 1500
aacaacgagt tcaaccagaa catcaaggac ctgctgccga aggacaggat cggcaacacc 1560
atcaacggcg agttcgcctt cgagggcgac gaggagagga agatcgagtt ctacaagacc 1620
gtgctgaact ccaagtacgc caagcaggtg ctgaacatcc cgttcaagca ggtggaggag 1680
gagatcatct cccagtcctt cgagaacctg tccgacttcc agatcgccct ggagaagatc 1740
tgctacagga ggttcgccat ctactccaac tacatcatct ccttcgacgc ccagatcttc 1800
gacatcacct ccctggacct gaagaacaac gagaagaaca acctgaacac ccacacccac 1860
atctggaggg acttctggaa ggacgagaac gagaagaaca acttcgacat caggctgaac 1920
ccggagatca ccatctccta caggaccccg aagcagtcca ggatcgagaa gtacggcgag 1980
aagaccaagg agtacgaccc gaacaagaac aacaggtacc tgcacccgca gttcaccctg 2040
atcaccacca tctccgagag gtccaactcc cagaccaaga ccctgtcctt catcgaggac 2100
gaggacttca agaagtccat caacgagttc aacaagaagc tgaagaagga caacatcaag 2160
ttcgccttcg gcatcgacaa cggcgaggtg gagctgtcca ccctgggcgt gtacctgccg 2220
accttcgaga aggagaccca cgaggagaag atctacgagc tgaagcagat caagaagtac 2280
ggcttcgagg tgctgaccat caccgacctg aagtacaagg agaccgacta caacggcaac 2340
gtgaggaaga tcatccagaa cccgtcctac ttcctgaaga aggagaacta catcaggacc 2400
ttctccaagt ccgagcagga gtacgaggag atgttcgcca agctgttcaa gaaggagcac 2460
gtgctgtccc tggacctgac caccgccaag atgatctgcg gccacatcgt gaccaacggc 2520
gacgtgccgg ccctgttcaa cctgtggctg aagcacgccc agaggaacgt gttcgagatg 2580
aacgaccaca ccgtgaagga gaccgccaag accatcaggc tgaggaacaa cgaggagctg 2640
accgacaacg agaaggagaa gttcgccgag ttcatctccg acggcaagaa gttcgccaag 2700
ctgaccaagg agggcaagaa gtccaggtac ctgaagtgga tcttcgagga caggaaggag 2760
aactccttca ccgaggacga gaacaagaag ttcaacgact gccagaagaa gaagggcaag 2820
tacaactccc acatcatctt cgcctccagg ttcgagggcg acgagctgaa gtccgtgacc 2880
ccgatcttcg actgcaggca cgtgttcaag aagaggaagg agttcgagac catcaggccg 2940
atcaaggaga tcgagaacga gatctccagg ttcaacacca acaggacctc ccacaacatc 3000
tccaacgagg agctggacct gaagatcacc gacgccaaga aggccctggt ggccaacgcc 3060
atcggcgtga tcgacttcct gtacaagcag tacaagcaga ggttcaacga cgagggcctg 3120
atcatcaagg agggcttcga cacccagaag gtggaggagg acatcgagaa gttctccggc 3180
aacatctaca ggatcctgga gaggaagctg taccagaagt tccagaacta cggcctggtg 3240
ccgccgatca agaacctgat ggccgtgagg aacgagggca tcaaggacaa gaacgccatc 3300
ctgaggctgg gcaacatcgc cttcatcgac ccgtccggca cctcccagga gtgcccggtg 3360
tgcaaggaga agtccaagga gaagcacacc aacaacttca tctgcgagtg cggcttcaac 3420
tccaccaaca tcatgcactc caacgacggc atcgccggct tcaacatcgc caagaggggc 3480
ttcgagaact tcatcaacga gaagtccagg gccgacccga agaagaagag gaaggtgtga 3540
<210> 111
<211> 3597
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 90所示融合蛋白的编码核酸序列
<400> 111
atggagaagt acaagatcac caagaccatc aggttcaggc tggacgccga caacaccgcc 60
atctccgcca tcgtgaagga caccgaggcc ctggaggcca ggggccaggg cttcaagatc 120
aagaagttcg tgaacgccct gggcaggttc ctgtccggcg acggcgtgca gaagtacctg 180
tacgacatgt ccaacgagga gaactgcgtg ttcaagagga acctggtgat caagaacacc 240
tggctgaaga acaacgccaa gcaggagatc gccggcatgg acctgaagag gggcctgatc 300
atcaaggaca tcaagggcct gcaggacaag atcgaggaga tctacgacaa gctgtgggag 360
atctacgaga tcctgtacga gtccgcctac ctgccgctgc aggacctggc caggagggag 420
ggcatcggcc tgctgctgaa gaagctgtcc gtgaagaacg ccctgccgtt catcatctcc 480
ttcgtggagg agtccaacga caagaacgag gccgacgacc tgtccctgag gctgaagaag 540
cagggcaagg agatcctgac ccagctggag atcggcatca acgagtacct gccggcccag 600
tcctccggcc tgccggtggc caaggcctcc ttcaactact acaccatcaa caagaccccg 660
gtggacttcg gcgagaagat ccaggagctg gagaagaggc tgtccgtgga catcaagaag 720
gagatctcct ccttcaccgg cggcatcaag accgccatca agaacaagat cgccggcaag 780
aagatcctgc tgggcgacac cccgatgttc gagtccgaga actccgtgtc cctgaggcag 840
atcctgaaga acatcaagtc cgagcagaag gcccagttca acaagttcat gaccacccag 900
aacaacccgc agctggagga gatgaagacc atgggctggt acctgttcgg cgacatcacc 960
gagggcgagt tcaacgacta caaggagcag accaaggaga tcgagagggt gggcgccaag 1020
atcaaccagt gcggcaacat caaggagaag aaggagctga ggtcccagct gcagaagctg 1080
aagaagaaga ggggcgagct gatctccgag gcccacaaga agggcggcaa cgacaagaac 1140
ttcaagacct acaaggagtt cgccaagttc tacaggaaga tcgcccagag gcacggcaag 1200
atcctggccc agatcaaggg catcgagaag gagaagatcg actccgccat gctgaactac 1260
tgggccgccg tgatcgagct gtccggcagg cacaagctgg tgctgatccc gaagaaggac 1320
gagaacgcca agaagtgcat cgagtggctg gaggacgagt ccaagcacaa gaacggctcc 1380
tgcaagatct tctggttcga gtccttcacc ttcaggtccc tgcagaagct gtgcttcggc 1440
aacctggact ccggcaccaa caccttcaac cagaagatcc agaacctgct gccgtgcgac 1500
gagaggggca acctgatgaa cggcgagttc gccttcaagg gcgacgagca ggagaagatc 1560
aagttctaca agaaggtgct gcagtcccag aaggacatca acctgccgca gaaggaggtg 1620
gtggacaacg tggtgggcag gaagttcgag accatggacg agttcaagat cgccctggag 1680
gagatctgct acatcaggag ggagaggctg tccgccaacg ccgagtccga gctgaagtcc 1740
aagttcaacg cccagatctt cgacatcacc tccctggacc tgaggaaccc ggtgaactgc 1800
gccggcaagc cggaggtgta ccaccacaac gacaagaggc acaccgagat ctggaaggag 1860
ttctggtccc tggacaacga gaggaggaac ttcaacatca ggctgaaccc ggagatcacc 1920
atcacctaca ggaagccgaa ggagtccagg atcctgaagt acggcaaggg caccgagaag 1980
tacaacgccg acatgaagaa caggtacctg tacccgcagt acaccctgct gaccaccatc 2040
tccgagcact gcaacacccc gaccaagatc ctgtccttca tgaccgacaa cgagtacgag 2100
gagtccatca aggccttcaa ctccaagctg aagaaggagg acatcaagtt cgccttcggc 2160
atcgactccg gcgagaccga gctgtccacc ctgggcgtgt acctgccgga gttctccgcc 2220
gagtccaccg agctgaagga catcgagaag tacggcttca acgtgctgac catcaaggac 2280
ctgaactaca ccgagaccga ctacaacggc tccgacaaga agatcgtgaa gaacccgtcc 2340
tacttcgtgg acaagtccct gtacatgagg accttcaaga agaccgagca ggagtacgag 2400
aagatgttcg ccgagcagtt cgaggccaag aagaggctgt ccctggacct gtccgccgcc 2460
aaggtgatct gcggccacat cgtgaccaac ggcggcgtgt ccgagcactt cggcctgtgg 2520
ctgaagcacg cccagaggac catcttctgg atgaacgacc acaccgagaa gaagaccgcc 2580
aagaacatca agctgaagga ctcctccgag ctgacctacg acgagaggga gaagttcgcc 2640
gagcacatct cctccgacga gaagttcaag aagctggacg tggaggagaa gaagaggtac 2700
gtgaggtgga tcttcgagga cagggagacc ctgaacttca ccgaggccga gaacaagaag 2760
ttcggcggct accagaagaa gaagggcgac tacaggctgg gcatcctgtt cgcctcctgc 2820
ttcatcggca aggagctgga gtccgtgacc cagatcctgg actgcaggca catcttcaag 2880
aagagggagg agttctactc cctgaagtcc aaggaggaca tcgaggccga gatcaagagg 2940
tacaacaccg actacaccaa ccacaacatc tccaccgagc agctggacct gaagttcgtg 3000
aacgtgaaga acgccctggt ggccaacgcc gtgggcgtga tcgacctgct gtacaagcag 3060
tacaaggaga ggctgggcgg cgagggcctg atcgccaagg agggcttcga caccaagaag 3120
gtggaggagg acatggagaa gttctccggc aacatctaca ggatcctgga gaggaagctg 3180
taccagaagt tccagaacta cggcctggtg ccgccgatca agaacctgat ggccgtgagg 3240
gccgacaagg tggagatctc cgaggccgag aagtccaaga tcagggagaa ctgcaagatc 3300
tccaagatcg acccggagaa cgagatcatc aagaggaaca agaccctgat cctgaggctg 3360
ggctccatcg ccttcgtgaa cgacgccgac acctcccagg agtgcccggc ctgcggcacc 3420
aagtccaagg agaagcacgt ggacaacttc atctgcggct gcggcttcaa ctccaccggc 3480
atcatccact ccaacgacgg cgtggccggc ttcaacatcg ccaagagggg cttcgtgaac 3540
ctgatggagc acgagctgag gtccagggcc gacccgaaga agaagaggaa ggtgtga 3597
<210> 112
<211> 3621
<212> DNA
<213> 人工序列
<220>
<223> SEQ ID NO: 91所示融合蛋白的编码核酸序列
<400> 112
atggagaagt acaagctgac caagaccatc aggttcaagc tgaagccgaa ggacatctcc 60
gccatcaaga gggacgtgga ggccctggag cagcagaagt tcgacctggt gctgttcgtg 120
tacaacctgc acaacttcat cggcaagctg aaggagtacc tgttcttcca gaaggagaag 180
gacgagttcg tgatcaagga caagctgacc atcaagaaga cctggctgaa gcagtacgcc 240
aagcaggaga tcgccggcct ggagctgaac agggagcaga ccctgggcaa catcaagggc 300
gtgtccgcca ggatcgagag ggccgtggac gacgtgaaca agatctacgt ggagctggcc 360
atggaggcca agctgaacga gagggccaag aaggccaaga ccgagcagct gatcaagagg 420
ctggacacca ggaacgccct gccgctgctg gtgtccctga tcgagcagtc ctccgacaag 480
tacgagaccg gcaacctgtc catccagctg aagaggctgg gcaagaggct gcagacccag 540
ctgctgtccg gcatcaagaa gtacctggcc gagcagtcca acggcctgcc gatcgccaag 600
gcctccttca actactacgc catcaacaag aagccggtgg actacatcga caagatcaag 660
cagctgcaga aggacctgga gatcaagaag aacaggaggt ccgaggagag gtacgacaag 720
aagaagagga agaacatcaa gatcttcaac gactccaagc tgtggatcaa gatcaagaag 780
gacatcgaga aggagagggg caacaagacc ctgatcctgg gctacgcccc gatgatcgag 840
ccgggcaact acgtgtacct gaggcagatc ctgaagaaca tcaagctgga gcagaagaac 900
aagttctcca agctgatgca gtccaagtcc ctgaccttcc acgacctgaa caacaacaac 960
cagctgtacc tgttcaagga catcctggag ggcgagttca acaagtacaa gcagaagacc 1020
aacgagatcg agaccaaggc cgagaagagg aaccagtgca acaacgacga gctgaagagg 1080
aagctgaact ccgagctgca gcagctgagg aaggacaggg gctccctgat caacgccgcc 1140
gacggcaggc cgaagggcag gttcaagacc tacaagtact tcgccaactt ctacaggaac 1200
gtggcccaga agcacggcag gatcctgtcc accctgaagg gcatcgagaa ggagatggtg 1260
gagtcccagc tgctgaagta ctggaccatc atcaccgagg agaacaacca gcactccctg 1320
gtgctgatcc cgaaggagag ggccggcgag tacaagaagg acctggagaa ctccatcccg 1380
tccgacccgt cctccaagat caaggtgtac tggttcgagt ccttcaccct gaggtccctg 1440
aggaagctgt gcttcggcta cgtgaacaac aacaccggct ccaacacctt ctacccggag 1500
ctgaagaagt ccgacgagct gaggaagtac cacgacgaga ggggcaactt catcaagggc 1560
gagttctact tcaagggcga cgagcagaag atcatccagt tctacaagga cgtgctgagg 1620
tccaactacg cccagaaggt gctgaagttc ccgaagcagc aggtgaagga cgagctgatc 1680
ggcagggagt tctcctccct ggacgagttc cagatcgccc tggagaagat ctgctaccag 1740
aggcacgtgg tgtgctccca gaaggtggtg gacgccctgt ccaggtacaa cgcccagatc 1800
ttcctgatca cctccctgga cctgggcaac ccggccaact gcgtggacaa gccgaagcag 1860
ttctcccact tcgacaagaa gcacaccagg atctggaagg agttctggtc ctccaagaac 1920
gagaccgcca acttcgacat caggctgaac ccggagatcg tgatcaccta caggcagccg 1980
aagcagtcca ggatcaagaa gtacggcccg gagtccacca ggtacgacga caggaagcac 2040
aacaggtacc tgtacccgca gttcaccctg atcaccacca tctccgagta ctccaacgcc 2100
ccgaccaagg ccctgtcctt cctgaccgac gaggagttca agggcgccgt ggacgagttc 2160
aacaagaagt tcaagaagga gaacatcagg ttctccctgg gcatcgacaa cggcgagacc 2220
gagctgtcca ccctgggcgt gtacctgccg gtgttcaaga aggactccaa cgagaaggtg 2280
gtggccgagc tgaagaaggt gaacaagtac ggcttcaact tcctgaccat caaggacctg 2340
tcccacgtgg agaaggacaa gaacggcagg gtgaggaaga tcatccagaa cccgtcctac 2400
ttcctgtcca aggagcagta catgaggacc ttcggcagga ccgagcagga gtacaacaac 2460
atgttcgccg agcagttcga ggagaaggcc ttcctgtccc tggacctgac caccgccaag 2520
gtgatcaacg gccacatcgt gaccaacggc gacgtgccga ccttcctgaa cctgtggatg 2580
aggcacgccc agagggacat ctgggacatg aacgaccaca ccaaggagaa gaccgccaag 2640
aagatcgtga tcaagaacaa cgacgagctg accgacgccg agaaggtgaa gttcgtggag 2700
tacatctccg acgagaccaa ctacgccaag ctgaacttca acgagaagaa gaggtacgtg 2760
ctgtggatct tcgagaacag gaagaacatc aacttcaccg acgccgagaa gaagaagttc 2820
gagccgtgcc agaagaggaa gggcaacttc tccaaggaca tcctgttcgc cgtgtgctac 2880
atcggctccg agatccactc cgtgaccaac atcttcgacg tgaggaacat cttcaagatg 2940
aggaaggact tctacgtgct gaagtccgag atggagatca agaaggagat cgagtcctac 3000
aacaccaccg ccggcatcca ggagatctcc aacgaggagc tggacctgaa gatcaacagg 3060
ctgaagcagg ccgtggtggc caacgccgtg ggcgtgatcg actacctgta catctactac 3120
aagaagaaga ccggcggcga gggcctgatc atcaaggagg gcttcgacac caagaaggtg 3180
gccaaggccc tggagaagtt ctccggcaac atctacagga tcctggagag gaagctgtac 3240
cagaagttcc agaactacgg cctggtgccg ccgatcaagt ccctgatggc cgtgagggag 3300
gagggcatcg agaacaacaa ggacgccatc ctgaggctgg gcaacgtggg cttcatcgac 3360
ccgaccggca cctcccagca gtgcccggtg tgctccaagg gcaagctgaa ccacaccacc 3420
aagtgctcca agaactgcgg cttcaactcc aagaacatca tgcactccaa cgacggcatc 3480
gccggctaca acatcgccaa gaggggcttc gagaacttca tctcccagaa gaagggctac 3540
gacgtgatca acaacggcac caagtacaac aacctgaagt cccagtccag ggccgacccg 3600
aagaagaaga ggaaggtgtg a 3621

Claims (91)

1.一种蛋白，其氨基酸序列如SEQ ID NO:3所示。
2.权利要求1所述的蛋白，其中，所述蛋白具有Cas效应蛋白活性。
3.权利要求1或2所述的蛋白，其中，所述蛋白是CRISPR/Cas***中的效应蛋白。
4.一种缀合物，其包含权利要求1-3任一项所述的蛋白以及修饰部分。
5.权利要求4所述的缀合物，其中，所述修饰部分选自另外的蛋白或多肽、可检测的标记，及其任意组合。
6.权利要求4所述的缀合物，其中，所述修饰部分通过接头连接至所述蛋白的N端或C端。
7.权利要求5所述的缀合物，其中，所述另外的蛋白或多肽选自表位标签、报告基因序列、核定位信号(NLS)序列、靶向部分、转录激活结构域、转录抑制结构域、核酸酶结构域，具有选自下列的活性的结构域：甲基化酶活性，去甲基化酶活性，转录激活活性，转录抑制活性，转录释放因子活性，组蛋白修饰活性，核酸酶活性和核酸结合活性；以及其任意组合。
8.权利要求7所述的缀合物，其中，所述核酸酶活性选自单链RNA切割活性，双链RNA切割活性，单链DNA切割活性，双链DNA切割活性。
9.权利要求7所述的缀合物，其中，所述转录激活结构域为VP64，所述转录抑制结构域为KRAB结构域或SID结构域，和/或，所述核酸酶结构域为Fok1。
10.权利要求4所述的缀合物，其中，所述缀合物包含表位标签。
11.权利要求4所述的缀合物，其中，所述缀合物包含NLS序列。
12.权利要求11所述的缀合物，其中，所述NLS序列如SEQ ID NO:73所示。
13.权利要求11所述的缀合物，其中，所述NLS序列位于所述蛋白的N端或C端。
14.一种融合蛋白，其包含权利要求1-3任一项所述的蛋白以及另外的蛋白或多肽。
15.权利要求14所述的融合蛋白，其中，所述另外的蛋白或多肽通过接头连接至所述蛋白的N端或C端。
16.权利要求14所述的融合蛋白，其中，所述另外的蛋白或多肽选自表位标签、报告基因序列、核定位信号(NLS)序列、靶向部分、转录激活结构域、转录抑制结构域、核酸酶结构域，具有选自下列的活性的结构域：甲基化酶活性，去甲基化酶活性，转录激活活性，转录抑制活性，转录释放因子活性，组蛋白修饰活性，核酸酶活性和核酸结合活性；以及其任意组合。
17.权利要求16所述的融合蛋白，其中，所述核酸酶活性选自单链RNA切割活性，双链RNA切割活性，单链DNA切割活性，双链DNA切割活性。
18.权利要求16所述的融合蛋白，其中，所述转录激活结构域为VP64，所述转录抑制结构域为KRAB结构域或SID结构域，和/或，所述核酸酶结构域为Fok1。
19.权利要求14所述的融合蛋白，其中，所述融合蛋白包含表位标签。
20.权利要求14所述的融合蛋白，其中，所述融合蛋白包含NLS序列。
21.权利要求20所述的融合蛋白，其中，所述NLS序列如SEQ ID NO:73所示。
22.权利要求20所述的融合蛋白，其中，所述NLS序列位于所述蛋白的N端或C端。
23.权利要求14所述的融合蛋白，其中，所述融合蛋白具有如SEQ ID NO:76所示的氨基酸序列。
24.一种分离的核酸分子，其由选自下列的序列组成：
(a)SEQ ID NO：39所示的核苷酸序列；
或
(b)SEQ ID NO：39所示的核苷酸序列的互补序列。
25.权利要求24所述的分离的核酸分子，其中，所述分离的核酸分子是RNA。
26.权利要求24所述的分离的核酸分子，其中，所述分离的核酸分子是CRISPR/Cas***中的同向重复序列。
27.一种复合物，其包含：
(i)蛋白组分，其选自：权利要求1-3任一项所述的蛋白、权利要求4-13任一项所述的缀合物、权利要求14-23任一项所述的融合蛋白，及其任意组合；和
(ii)核酸组分，其从5’至3’方向包含权利要求24-26任一项所述的分离的核酸分子和能够与靶序列杂交的导向序列，
其中，所述蛋白组分与核酸组分相互结合形成复合物。
28.权利要求27所述的复合物，其中，所述导向序列连接于所述核酸分子的3’端。
29.权利要求27所述的复合物，其中，所述导向序列包含所述靶序列的互补序列。
30.权利要求27所述的复合物，其中，所述核酸组分是CRISPR/Cas***中的导向RNA。
31.权利要求27所述的复合物，其中，所述核酸分子是RNA。
32.权利要求27所述的复合物，其中，所述复合物不包含反式作用crRNA(tracrRNA)。
33.一种分离的核酸分子，其包含：
(i)编码权利要求1-3任一项所述的蛋白、或权利要求14-23任一项所述的融合蛋白的核苷酸序列；
(ii)编码权利要求24-26任一项所述的分离的核酸分子的核苷酸序列；和/或，
(iii)包含(i)和(ii)的核苷酸序列。
34.权利要求33所述的分离的核酸分子，其中，(i)-(iii)任一项中所述的核苷酸序列经密码子优化用于在原核细胞或真核细胞中进行表达。
35.一种载体，其包含权利要求33或34所述的分离的核酸分子。
36.一种宿主细胞，其包含权利要求33或34所述的分离的核酸分子或权利要求33所述的载体。
37.一种组合物，其包含：
(i)第一组分，其选自：权利要求1-3任一项所述的蛋白、权利要求4-13任一项所述的缀合物、权利要求14-23任一项所述的融合蛋白、编码所述蛋白或融合蛋白的核苷酸序列，以及其任意组合；和
(ii)第二组分，其为包含导向RNA的核苷酸序列，或者编码所述包含导向RNA的核苷酸序列的核苷酸序列；
其中，所述导向RNA从5’至3’方向包含同向重复序列和导向序列，所述导向序列能够与靶序列杂交；
所述导向RNA能够与(i)中所述的蛋白、缀合物或融合蛋白形成复合物。
38.权利要求37所述的组合物，其中，所述同向重复序列是权利要求24-26任一项中所定义的分离的核酸分子。
39.权利要求37所述的组合物，其中，所述导向序列连接至所述同向重复序列的3’端。
40.权利要求37所述的组合物，其中，所述导向序列包含所述靶序列的互补序列。
41.权利要求37所述的组合物，其中，所述组合物不包含反式作用crRNA(tracrRNA)。
42.权利要求37所述的组合物，其中，所述组合物中的至少一个组分是非天然存在的或经修饰的。
43.一种组合物，其包含一种或多种载体，所述一种或多种载体包含：
(i)第一核酸，其为编码权利要求1-3任一项所述的蛋白或权利要求14-23任一项所述的融合蛋白的核苷酸序列；任选地所述第一核酸可操作地连接至第一调节元件；以及
(ii)第二核酸，其编码包含导向RNA的核苷酸序列；任选地所述第二核酸可操作地连接至第二调节元件；
其中：
所述第一核酸与第二核酸存在于相同或不同的载体上；
所述导向RNA从5’至3’方向包含同向重复序列和导向序列，所述导向序列能够与靶序列杂交；
所述导向RNA能够与(i)中所述的蛋白或融合蛋白形成复合物。
44.权利要求43所述的组合物，其中，所述同向重复序列是权利要求24-26任一项中所定义的分离的核酸分子。
45.权利要求43所述的组合物，其中，所述导向序列连接至所述同向重复序列的3’端。
46.权利要求43所述的组合物，其中，所述导向序列包含所述靶序列的互补序列。
47.权利要求43所述的组合物，其中，所述组合物不包含反式作用crRNA(tracrRNA)。
48.权利要求43所述的组合物，其中，所述组合物中的至少一个组分是非天然存在的或经修饰的。
49.权利要求43所述的组合物，其中，所述第一调节元件和/或所述第二调节元件是启动子。
50.权利要求49所述的组合物，其中，所述启动子是诱导型启动子。
51.权利要求37或43所述的组合物，其中，当所述靶序列为DNA时，所述靶序列位于原间隔序列临近基序(PAM)的3’端，并且所述PAM具有5’-TG所示的序列；当所述靶序列为RNA时，所述靶序列不具有PAM结构域限制。
52.权利要求37或43所述的组合物，其中，所述靶序列是来自原核细胞或真核细胞的DNA或RNA序列；或者，所述靶序列是非天然存在的DNA或RNA序列。
53.权利要求37或43所述的组合物，其中，所述靶序列存在于细胞内；或者，所述靶序列存在于体外的核酸分子中。
54.权利要求53所述的组合物，其中，所述靶序列存在于细胞核内或细胞质内；或者，所述靶序列存在于质粒中。
55.权利要求53所述的组合物，其中，所述细胞是真核细胞或原核细胞。
56.权利要求37所述的组合物，其中，所述蛋白连接有一个或多个NLS序列，或者，所述缀合物或融合蛋白包含一个或多个NLS序列。
57.权利要求56所述的组合物，其中，所述NLS序列连接至所述蛋白的N端或C端。
58.一种试剂盒，其包括一种或多种选自下列的组分：权利要求1-3任一项所述的蛋白、权利要求4-13任一项所述的缀合物、权利要求14-23任一项所述的融合蛋白、权利要求24-26任一项所述的分离的核酸分子、权利要求27-32任一项所述的复合物、权利要求33或34所述的分离的核酸分子、权利要求35所述的载体、权利要求37-57任一项所述的组合物。
59.权利要求58所述的试剂盒，其中，所述试剂盒包含权利要求37-42、51-57任一项所述的组合物，以及使用所述组合物的说明书。
60.权利要求58所述的试剂盒，其中，所述试剂盒包含权利要求43-57任一项所述的组合物，以及使用所述组合物的说明书。
61.一种递送组合物，其包含递送载体，以及选自下列的一种或多种：权利要求1-3任一项所述的蛋白、权利要求4-13任一项所述的缀合物、权利要求14-23任一项所述的融合蛋白、权利要求24-26任一项所述的分离的核酸分子、权利要求27-32任一项所述的复合物、权利要求33或34所述的分离的核酸分子、权利要求35所述的载体、权利要求37-57任一项所述的组合物。
62.权利要求61所述的递送组合物，其中，所述递送载体是粒子。
63.权利要求61所述的递送组合物，其中，所述递送载体选自脂质颗粒、糖颗粒、金属颗粒、蛋白颗粒、脂质体、外泌体、微泡或病毒载体。
64.权利要求63所述的递送组合物，其中，所述病毒载体为复制缺陷型逆转录病毒、慢病毒、腺病毒或腺相关病毒。
65.一种修饰靶基因的方法，其包括：将权利要求27-32任一项所述的复合物或权利要求37-57任一项所述的组合物与所述靶基因接触，或者递送至包含所述靶基因的细胞中；所述靶序列存在于所述靶基因中；并且，所述方法用于非治疗目的。
66.权利要求65所述的方法，其中，所述靶基因存在于细胞内，或者，所述靶基因存在于体外的核酸分子中。
67.权利要求66所述的方法，其中，所述靶基因存在于质粒中。
68.权利要求66所述的方法，其中，所述细胞是原核细胞或真核细胞。
69.权利要求66所述的方法，其中，所述细胞选自哺乳动物细胞、植物细胞。
70.权利要求66所述的方法，其中，所述细胞为人类细胞。
71.权利要求65所述的方法，其中，所述修饰是指所述靶序列的断裂。
72.权利要求71所述的方法，其中，所述靶序列的断裂为DNA的双链断裂或RNA的单链断裂。
73.权利要求71所述的方法，其中，所述修饰还包括将外源核酸***所述断裂中。
74.一种改变基因产物的表达的方法，其包括：将权利要求27-32任一项所述的复合物或权利要求37-57任一项所述的组合物与编码所述基因产物的核酸分子接触，或者递送至包含所述核酸分子的细胞中，所述靶序列存在于所述核酸分子中；并且，所述方法用于非治疗目的。
75.权利要求74所述的方法，其中，所述核酸分子存在于细胞内，或者所述核酸分子存在于体外的核酸分子中。
76.权利要求75所述的方法，其中，所述核酸分子存在于质粒中。
77.权利要求75所述的方法，其中，所述细胞是原核细胞或真核细胞。
78.权利要求75所述的方法，其中，所述细胞选自哺乳动物细胞、植物细胞。
79.权利要求75所述的方法，其中，所述细胞为人类细胞。
80.权利要求74所述的方法，其中，所述基因产物的表达增强或降低。
81.权利要求74所述的方法，其中，所述基因产物是蛋白。
82.权利要求74所述的方法，其中所述的复合物或组合物包含于递送载体中。
83.权利要求82所述的方法，其中，所述递送载体选自脂质颗粒、糖颗粒、金属颗粒、蛋白颗粒、脂质体、外泌体、病毒载体。
84.权利要求83所述的方法，其中，所述病毒载体为复制缺陷型逆转录病毒、慢病毒、腺病毒或腺相关病毒。
85.权利要求74-84任一项所述的方法，其用于改变靶基因或编码靶基因产物的核酸分子中的一个或多个靶序列来修饰细胞、细胞系或生物体。
86.一种体外的、离体的细胞或细胞系或它们的子代，所述细胞或细胞系或它们的子代包含：重组的权利要求1-3任一项所述的蛋白、权利要求4-13任一项所述的缀合物、权利要求14-23任一项所述的融合蛋白、权利要求24-26任一项所述的分离的核酸分子、权利要求27-32任一项所述的复合物、权利要求33或34所述的分离的核酸分子、权利要求35所述的载体、权利要求37-57任一项所述的组合物。
87.权利要求86所述的细胞或细胞系或它们的子代，其中，所述细胞是原核细胞或真核细胞。
88.权利要求1-3任一项所述的蛋白、权利要求4-13任一项所述的缀合物、权利要求14-23任一项所述的融合蛋白、权利要求24-26任一项所述的分离的核酸分子、权利要求27-32任一项所述的复合物、权利要求33或34所述的分离的核酸分子、权利要求35所述的载体、权利要求37-57任一项所述的组合物或权利要求58-60任一项所述的试剂盒，用于核酸编辑的用途，或者在制备制剂中的用途，所述制剂用于核酸编辑；并且，所述用途为非治疗用途。
89.权利要求88所述的用途，其中，所述核酸编辑包括基因或基因组编辑。
90.权利要求89所述的用途，其中，所述基因或基因组编辑包括修饰基因、敲除基因、改变基因产物的表达、修复突变、和/或***多核苷酸。
91.权利要求1-3任一项所述的蛋白、权利要求4-13任一项所述的缀合物、权利要求14-23任一项所述的融合蛋白、权利要求24-26任一项所述的分离的核酸分子、权利要求27-32任一项所述的复合物、权利要求33或34所述的分离的核酸分子、权利要求35所述的载体、权利要求37-57任一项所述的组合物或权利要求58-60任一项所述的试剂盒，在制备制剂中的用途，所述制剂用于：
(i)体外或离体单链DNA的检测；
(ii)编辑靶基因座中的靶序列来修饰非人类生物。
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