CN112703250A - CRISPRi在高通量代谢工程中的应用 - Google Patents

CRISPRi在高通量代谢工程中的应用 Download PDF

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CN112703250A
CN112703250A CN201980060677.5A CN201980060677A CN112703250A CN 112703250 A CN112703250 A CN 112703250A CN 201980060677 A CN201980060677 A CN 201980060677A CN 112703250 A CN112703250 A CN 112703250A
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B·柴金德
H·M·范罗苏姆
A·米勒
P·珀科维奇
S·希捷卡
K·帕特尔
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Abstract

本公开描述了用于体外高通量DNA组装反应的方法、组合物和试剂盒。本公开进一步描述了模块化CRISPR DNA构建体，所述模块化CRISPR DNA构建体包括模块化插入DNA部分，所述模块化插入DNA部分侧接有包括预先验证的CRISPR原间隔子/原间隔子邻近基序序列组合的克隆标签区段。还公开了CRISPRi和CRISPRa的高通量方法。

Description

CRISPRi在高通量代谢工程中的应用
相关申请的交叉引用
本申请要求于2018年8月15日提交的美国临时申请第62/764,672号的优先权的权益，所述美国临时申请出于所有目的通过全文引用的方式并入在此，包含所有描述、参考文献、附图和权利要求。
政府资助
本发明是根据DARPA授予的协议号HR0011-15-9-0014在政府支持下进行的。政府拥有本发明的某些权利。
关于序列表的声明
与本申请相关的序列表以文本格式提供以代替纸质副本并且在此通过引用的方式并入本说明书。含有序列表的文本文件的名称是ZYMR_030_01WO_SeqList_ST25.txt。文本文件为799kb、创建于2019年8月14日并且通过EFS-Web以电子方式提交。
技术领域
本公开涉及用于在体外进行引导的基因序列编辑的系统、方法和组合物。本公开尤其描述了使用引导的序列编辑复合物用于改进的DNA克隆、寡核苷酸的组装和微生物的改进的方法。本公开还描述了通过突变CRISPR酶调节宿主细胞基因的表达的高通量方法。
背景技术
生物学中的主要感兴趣领域是体外和体内靶向的基因序列修饰。事实上，学术和商业基因研究的最重要瓶颈之一一直是在测试之前可以生成或稍后修饰新基因构建体的速度。
对于稍后的修饰，依赖于限制性位点识别或DNA杂交和扩增的当前可用克隆技术已被证明是缓慢、不可靠且难处理的。成簇规律间隔短回文重复序列(CRISPR)基因编辑系统的发现已经为研究人员提供了用于基因修饰的另外的途径。然而，即使是这些新方法对于高通量模块化克隆应用来说仍然是不切实际的。
例如，催化活性或灭活CRISPR酶的使用允许研究人员实现靶向的基因表达阻遏(CRISPRi)或激活(CRISPRa)。但是，这些技术仍然受到CRISPR应用的技术限制并且因此未针对高通量应用进行优化。
例如，CRISPR编辑定位通常受到原间隔子邻近基序(PAM)的定位的限制。从头CRISPR引导RNA设计和基因靶向可以是耗时且昂贵的并且也易受低效率影响并且可能发生脱靶突变。
因此，需要用于基因序列的靶向的改变和基因表达的调节的改进的组合物和方法。
发明内容
在一些实施例中，本公开教导了用于利用模块化CRISPR DNA构建体的高通量体内和体外DNA组装反应的方法、组合物和试剂盒。
因此，在一些实施例中，本公开教导了模块化CRISPR DNA构建体，所述模块化CRISPR DNA构建体包括模块化插入DNA部分，所述模块化插入DNA部分侧接有包括预先验证的CRISPR原间隔子/原间隔子邻近基序(PAM)序列组合的克隆标签区段。在一些实施例中，本公开教导了用CRISPR核酸内切酶消化DNA。在一些实施例中，本公开教导了用II型-2类CRISPR核酸内切酶(例如，Cas9)来消化DNA。在一些实施例中，本公开教导了用V型-2类CRISPR核酸内切酶消化DNA。在一些实施例中，本公开教导了用Cpf1核酸内切酶消化DNA。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，所述重组模块化CRISPR DNA构建体包括CRISPR多克隆位点，所述多克隆位点包括：a)至少两个不同的克隆标签(cTAG)，其中每个cTAG包括：i)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序(PAM)可操作地连接的原间隔子序列；其中所述经过验证的CRISPR着陆位点中的至少一个在所述模块化CRISPR DNA构建体中是独特的；以及b)一或多个DNA插入序列；i)其中所述cTAG中的每个cTAG围绕所述一或多个DNA插入序列中的每个DNA插入序列分布在侧翼位置；并且ii)其中所述DNA插入序列中的至少一个包括选择标志物。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体是环状的。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体是线性的。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体整合到生物体的基因组中。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述不同的cTAG中的至少一个包括至少两个经过验证的CRISPR着陆位点。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述CRISPR着陆位点中的至少一个是用于Cas9核酸内切酶。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述CRISPR着陆位点中的至少一个是用于RNA或DNA引导的核酸酶，例如Cpf1核酸内切酶。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述不同的cTAG中的至少一个包括罕见的(≥8个碱基长)限制性核酸内切酶位点。
在一些方面，本公开将重组模块化CRISPR DNA构建体称为“大模块化(MegaModular)”构建体。
在一些实施例中，本公开教导了一种用于制备重组核酸分子的方法，所述方法包括：a)形成混合物，所述混合物包括：i)多个DNA插入部分，其中每个DNA插入部分侧接有两个克隆标签(cTAG)，每个cTAG包括：1)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序(PAM)可操作地连接的原间隔子序列；ii)一或多种CRISPR复合物，所述一或多种CRISPR复合物靶向所述多个DNA插入部分中的至少两个中存在的所述cTAG中的至少一个，每种CRISPR复合物包括：1)CRISPR核酸内切酶，以及2)能够将所述CRISPR核酸内切酶募集到所述靶向的cTAG之一的一或多个引导RNA；其中将所述混合物在允许消化所述多个DNA插入部分中的至少两个中的所述一或多个靶向的cTAG的条件下温育以产生突出端；以及b)在允许相容突出端的杂交和杂交的末端的共价连接的条件下温育(a)中生成的消化产物；其中所得重组核酸分子包括所述方法中连接的原始插入部分的完整cTAG序列。
在一些实施例中，本公开教导了一种用于制备重组核酸分子的方法，其中所述CRISPR核酸内切酶是Cpf1。
在一些实施例中，本公开教导了一种用于制备重组核酸分子的方法，其中所述CRISPR核酸内切酶是DNA或RNA引导的核酸内切酶，例如Cas9。
在一些实施例中，本公开教导了一种用于制备重组核酸分子的方法，其中所述方法包括以下步骤：i)在连接前将消化的cTAG序列与所述CRISPR复合物分离，或ii)在连接前灭活所述CRISPR复合物。
在一些实施例中，本公开教导了一种用于制备重组核酸分子的方法，其中分离步骤包括DNA纯化步骤。
在一些实施例中，本公开教导了一种用于制备重组核酸分子的方法，其中灭活步骤包括所述CRISPR复合物的热或化学灭活。
在一些实施例中，本公开教导了一种用于制备重组核酸分子的方法，其中所述多个DNA插入部分的每个DNA插入部分的所述两个cTAG形成cTAG对，并且其中所述cTAG对相对于所述方法中连接的所述DNA插入部分的所有其它cTAG是独特的。
在一些实施例中，本公开教导了一种用于制备重组核酸分子的方法，其中每个cTAG对中的所述cTAG中的至少一个与不同的cTAG对中的至少另一个cTAG相同。
在一些实施例中，本公开教导了一种用于DNA序列编辑的方法，所述方法包括：a)将以下引入反应中：i)本公开的模块化CRISPR DNA构建体：ii)置换DNA插入部分，其中所述置换DNA插入部分侧接有第一插入cTAG和第二插入cTAG；1)其中所述第一插入cTAG包括所述模块化CRISPR DNA构建体的所述不同cTAG中的一个的一或多个经过验证的CRISPR着陆位点，并且所述第二插入cTAG包括所述模块化CRISPR DNA构建体的另一个不同cTAG的一或多个经过验证的CRISPR着陆位点；以及iii)第一CRISPR复合物和第二CRISPR复合物，所述第一CRISPR复合物和所述第二CRISPR复合物分别靶向所述第一插入cTAG和所述第二插入cTAG，每种CRISPR复合物包括：1)CRISPR核酸内切酶，以及2)能够将所述CRISPR核酸内切酶募集到所述靶向的插入cTAG之一的引导RNA；其中所述第一CRISPR复合物和所述第二CRISPR复合物切割所述第一插入cTAG和所述第二插入cTAG以及它们对应的不同cTAG以生成突出端；以及b)温育所述置换DNA插入部分和具有生成的消化的cTAG的模块化CRISPRDNA构建体：(a)在允许相容突出端的杂交和杂交的末端的共价连接的条件下；其中所得经过编辑的模块化CRISPR DNA构建体包括通过所述方法连接的原始插入部分的完整cTAG序列。
在一些实施例中，本公开教导了一种用于DNA序列编辑的方法，其中步骤(b)的所述反应包括功能连接酶。
在一些实施例中，本公开教导了一种用于DNA序列编辑的方法，所述方法包括：a)将以下引入反应中：i)本公开的模块化CRISPR DNA构建体；ii)至少两种CRISPR复合物，所述至少两种CRISPR复合物靶向所述模块化CRISPR DNA构建体中的两个不同cTAG，每种CRISPR复合物包括：1)CRISPR核酸内切酶，以及2)能够将所述CRISPR核酸内切酶募集到所述靶向的不同cTAG之一的引导RNA；其中第一CRISPR复合物和第二CRISPR复合物切割所述模块化CRISPR DNA构建体中的所述两个不同cTAG，其中所得的不同cTAG包括突出端；以及b)将以下引入第二反应中：i)具有在(a)中生成的消化的cTAG的所述模块化CRISPR DNA构建体；以及ii)置换DNA插入部分，其中所述置换DNA插入部分侧接有第一插入cTAG和第二插入cTAG；1)其中所述第一插入cTAG包括在(a)中切割的未消化的不同cTAG之一的多核苷酸序列，并且所述第二插入cTAG包括在(a)中切割的另一个未消化的不同cTAG的多核苷酸序列；并且2)其中所述第一插入cTAG和所述第二插入cTAG包括与来自(a)的所述不同cTAG的所述突出端相容的突出端；所述第二反应是在允许相容的所述突出端的杂交和杂交的末端的共价连接的条件下；其中所得经过编辑的模块化CRISPR DNA构建体包括在(a)中靶向的原始未消化的不同cTAG的完整序列。
在一些实施例中，本公开教导了一种用于DNA序列编辑的方法，其中步骤(b)的所述反应包括功能连接酶。
在一些实施例中，本公开教导了一种用于DNA序列编辑的方法，其中所述CRISPR核酸内切酶是Cpf1。
在一些实施例中，本公开教导了一种用于DNA序列编辑方法，其中步骤(a)进一步包括用单链核酸外切酶消化两个切割的不同cTAG，由此产生具有突出端的所述不同cTAG。在一些方面，在核酸外切酶步骤后可以添加连接酶和聚合酶以用聚合酶和连接酶修复接合。在一些方面，此反应也可以用Cas9消化的平端切割来完成。
在一些实施例中，本公开教导了一种用于DNA序列编辑的方法，其中所述CRISPR核酸内切酶是Cas9。
在一些实施例中，本公开提供了一种宿主细胞基因组，所述宿主细胞基因组包括重组模块化CRISPR DNA构建体，所述重组模块化CRISPR DNA构建体包括CRISPR多克隆位点，所述多克隆位点包括：a)至少两个不同的克隆标签(cTAG)，其中每个cTAG包括：i)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序(PAM)可操作地连接的原间隔子序列；其中所述经过验证的CRISPR着陆位点中的至少一个在所述模块化CRISPR DNA构建体中是独特的；以及b)一或多个DNA插入部分；i)其中所述不同的cTAG中的每个cTAG围绕所述一或多个DNA插入部分中的每个DNA插入部分分布在侧翼位置。
在一些实施例中，本公开提供了一种用于制备重组核酸分子的方法，所述方法包括：a)温育混合物，所述混合物包括：i)多个DNA插入部分，所述多个DNA插入部分侧接有两个克隆标签(cTAG)，每个cTAG包括：1)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序(PAM)可操作地连接的原间隔子序列；以及2)罕见的(≥8个碱基)限制性酶识别位点；其中至少两个插入部分的所述cTAG中的至少一个包括相同的限制性酶位点；ii)一或多种限制性酶，所述一或多种限制性酶靶向所述多个DNA插入部分中的至少两个中的所述罕见的限制性酶位点；所述温育是在允许通过所述多个DNA插入部分中的至少两个中的所述一或多种限制性酶来消化靶向的cTAG以生成具有消化的DNA末端的插入部分的条件下；以及b)在允许消化的DNA末端共价连接的条件下温育具有步骤(a)中生成的所述消化的DNA末端的一或多个DNA插入部分；其中所得重组核酸分子包括所述方法中共价连接的原始插入部分的完整cTAG序列。
在一些实施例中，本公开提供了一种用于DNA序列编辑的方法，所述方法包括：a)提供以下：i)根据权利要求1所述的模块化CRISPR DNA构建体，其中所述不同的cTAG中的至少两个包括罕见的(≥8个碱基长)限制性酶识别位点；ii)置换DNA插入部分，其中所述置换DNA插入部分侧接有第一插入cTAG和第二插入cTAG；1)其中所述第一插入cTAG包括所述模块化CRISPR DNA构建体的所述不同cTAG中的一个的所述罕见的限制性酶识别位点，并且所述第二插入cTAG包括所述模块化CRISPR DNA构建体的另一个不同cTAG的所述罕见的限制性酶识别位点；以及iii)一或多种限制性酶，所述一或多种限制性酶靶向所述第一插入cTAG和所述第二插入cTAG中的所述罕见的限制性酶位点；其中部分(i)和(ii)各自在单个或单独的反应中与部分(iii)温育；其中所述一或多种限制性酶切割所述第一插入cTAG和所述第二插入cTAG以及它们对应的不同cTAG的所述罕见的限制性酶识别位点以生成消化的DNA末端；以及b)在允许所述消化的DNA末端共价连接的条件下温育所述置换DNA插入部分和具有步骤(a)中生成的消化的DNA末端的所述模块化CRISPR DNA构建体；其中所得经过编辑的模块化CRISPR DNA构建体包括通过所述方法共价连接的原始插入部分的完整cTAG序列。
在一些实施例中，本公开提供了一种用于制备重组核酸分子的方法，所述方法包括：a)温育混合物，所述混合物包括：i)多个DNA插入部分，其中每个DNA插入部分侧接有两个克隆标签(cTAG)，每个cTAG包括：1)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序(PAM)可操作地连接的原间隔子序列；其中所述DNA插入部分中的至少两个共享相同的cTAG；ii)单链DNA(ssDNA)核酸外切酶；所述温育是在允许所述至少两个DNA插入部分中的共享cTAG的消化的条件下，由此在所述至少两个DNA插入部分中生成相容的DNA突出端；以及b)在允许所述至少两个DNA插入部分的所述相容的DNA突出端的杂交和共价连接的条件下温育具有步骤(a)中生成的消化的cTAG的所述DNA插入部分；其中所得重组核酸分子包括消化之前的共享cTAG的完整cTAG序列。此反应也可以用用于固定接合的聚合酶和或连接酶来进行。进一步地，这可以用预先消化的载体来进行。
在一些实施例中，本公开教导了一种用于调节宿主细胞基因的表达或工程化宿主细胞的基因组的重组模块化CRISPR DNA构建体，所述构建体包括CRISPR多克隆位点，所述多克隆位点包括：a)至少两个不同的克隆标签(cTAG)，其中每个cTAG包括：i)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序(PAM)可操作地连接的原间隔子序列；其中所述经过验证的CRISPR着陆位点中的至少一个在所述模块化CRISPR DNA构建体中是独特的；以及b)一或多个DNA插入部分；i)其中所述不同的cTAG中的每个cTAG围绕所述一或多个DNA插入部分中的每个DNA插入部分分布在侧翼位置；并且其中所述一或多个DNA插入部分包括用于CRISPR功能调节剂的DNA。
在一些实施例中，本公开的重组模块化CRISPR DNA构建体包括针对CRISPR功能调节剂进行编码的DNA进一步包括可选择标志物。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述CRISPR功能调节剂选自由以下组成的组：复制起点、可选择标志物、可逆选择标志物、抗CRISPR蛋白、启动子、终止子、dCas蛋白、dCpf1蛋白、条形码、Cas9蛋白、Cpf1蛋白、DNA供体和促进多重化的蛋白质。
在一些实施例中，本公开教导了一种宿主细胞，所述宿主细胞包括如本说明书中所述的重组模块化CRISPR DNA构建体。
在一些实施例中，本公开教导了一种宿主细胞，其中所述宿主细胞包括对催化活性CRISPR酶进行编码的核酸分子和能够将催化激活CRISPR酶募集到DNA靶位点的引导RNA。在一些实施例中，本公开教导了一种宿主细胞，其中所述宿主细胞包括对催化灭活CRISPR酶进行编码的核酸分子和能够将所述催化灭活CRISPR酶募集到DNA靶位点的引导RNA。
在一些实施例中，本公开教导了一种宿主细胞，其中所述催化灭活CRISPR酶与转录激活蛋白融合。
在一些实施例中，本公开教导了一种宿主细胞，其中所述宿主细胞进一步包括对转录激活蛋白进行编码的核酸分子，所述转录激活蛋白在表达时能够将自身与所述催化灭活CRISPR酶连接。
在一些实施例中，本公开教导了一种宿主细胞，其中所述转录激活蛋白经由连接适配子或通过蛋白质-蛋白质相互作用将自身与所述催化灭活CRISPR酶连接。
在一些实施例中，本公开教导了一种宿主细胞，其中所述引导RNA与能够将自身与转录激活蛋白连接的适配子可操作地连接。
在一些实施例中，本公开教导了一种宿主细胞，其中所述转录激活蛋白选自由以下组成的组：VP16、VP64和VP160。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体是环状的。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体是线性的。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体整合到生物体的基因组中。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述不同的cTAG中的至少一个包括至少两个经过验证的CRISPR着陆位点。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述CRISPR着陆位点中的至少一个是用于Cas9核酸内切酶。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述CRISPR着陆位点中的至少一个是用于Cpf1核酸内切酶。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述不同的cTAG中的至少一个包括罕见的(≥8个碱基长)限制性核酸内切酶位点。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述催化灭活CRISPR酶是突变的Cas9核酸内切酶。
在一些实施例中，本公开教导了一种重组模块化CRISPR DNA构建体，其中所述催化灭活CRISPR酶是突变的Cpf1核酸内切酶。
在一些实施例中，本公开教导了一种宿主细胞，其中所述宿主细胞包括多于一个核酸引导RNA。
在一些实施例中，本公开教导了一种宿主细胞，其中所述引导RNA中的至少一个包括与另一个引导RNA不同的序列。
在一些实施例中，本公开教导了一种宿主细胞，其中所述引导RNA中的至少一个靶向与另一个引导RNA不同的DNA靶位点序列。
在一些实施例中，本公开教导了一种宿主细胞，其中所述宿主细胞包括多于一种催化灭活CRISPR酶。
在一些实施例中，本公开教导了一种宿主细胞，其中所述催化灭活CRISPR酶中的至少一种包括与所述构建体中编码的另一种催化灭活CRISPR酶不同的序列。
在一些实施例中，本公开教导了插入部分，其中所述cTAG中的一或多个选自由SEQID NO:65-74、78-81和其组合组成的组。
在一些实施例中，本公开教导了一种调节一或多个宿主细胞基因的表达的高通量方法，所述方法包括将本公开的重组模块化CRISPR DNA构建体引入宿主细胞中的步骤；其中引导RNA的DNA靶位点位于宿主细胞基因组内。
在一些实施例中，本公开教导了一种调节一或多个宿主细胞基因的表达的高通量方法，其中所述重组模块化CRISPR DNA构建体的至少一个插入部分整合到宿主细胞的基因组中。
在一些实施例中，本公开教导了一种调节一或多个宿主细胞基因的表达的高通量方法，其中所述插入部分调节CRISPR蛋白的功能。
在一些实施例中，本公开教导了一种调节一或多个宿主细胞基因的表达的高通量方法，其中所述插入部分调节gRNA的功能。
在一些实施例中，本公开教导了一种调节一或多个宿主细胞基因的表达的高通量方法，其中所述重组模块化CRISPR DNA构建体作为染色体外DNA保留在宿主细胞中。
在一些实施例中，本公开教导了一种用于筛选CRISPR酶变体的重组模块化CRISPRDNA构建体，所述构建体包括CRISPR多克隆位点，所述多克隆位点包括：a)至少两个不同的克隆标签(cTAG)，其中每个cTAG包括：i)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序(PAM)可操作地连接的原间隔子序列；其中所述经过验证的CRISPR着陆位点中的至少一个在所述模块化CRISPR DNA构建体中是独特的；以及b)一或多个DNA插入部分；i)其中所述不同的cTAG中的每个cTAG围绕所述一或多个DNA插入部分中的每个DNA插入部分分布在侧翼位置；其中所述构建体进一步包括：c)第一核酸，所述第一核酸对CRISPR酶或疑似具有CRISPR功能的酶(“推定的CRISPR酶”)进行编码；以及d)第二核酸，所述第二核酸对能够与DNA靶位点结合的引导RNA进行编码。
在一些实施例中，本公开教导了一种在宿主细胞中筛选CRISPR活性的高通量方法，所述方法包括以下步骤：a)将本公开的重组模块化CRISPR DNA构建体引入所述宿主细胞中；其中引导RNA的所述DNA靶位点位于宿主细胞基因组内；以及b)测量在所述DNA靶位点处发生的DNA切割的程度。
在一些实施例中，本公开教导了一种在宿主细胞中筛选CRISPRi和/或CRISPRa活性的高通量方法，所述方法包括以下步骤：a)将本公开的重组模块化CRISPR DNA构建体引入所述宿主细胞中；其中引导RNA的所述DNA靶位点位于宿主细胞基因组内；以及b)测量在所述DNA靶位点处发生的转录调节的程度。
附图说明
图1A-C展示了本公开的CRISPR/Cas9和CRISPR/Cpf1系统的比较。图1A—Cas9核酸内切酶通过tracrRNA和crRNA复合物募集到靶dsDNA。图1B—Cas9核酸内切酶也可以通过被称为单引导RNA(sgRNA)的人工融合的tracrRNA和crRNA序列募集到靶dsDNA。Cas9核酸内切酶产生平端。图1C—Cpf1核酸内切酶仅需要crRNA引导多核糖核苷酸。Cpf1核酸内切酶切割产生具有5'突出端的双链断裂。
图2A-C展示了利用本公开的模块化CRISPR构建体的本发明克隆方法的实施例。图2A—图解了根据本公开的可以用Cas9或Cpf1核酸酶容易地改变的模块化CRISPR质粒。如上所述，本公开的模块化CRISPR构建体可以称为“大模块化”构建体。由数字表示的可互换部分侧接有由字母表示的不变的cTAG序列。各部分可以预先组装或者可以基于cTAG序列同一性在体外组装。示例插入部分示出在图2A的右侧。图2B—可以使用如cTAG处的Cas9、Cpf1或限制性核酸内切酶切割等几种策略来置换单独部分而不必重新组装整个质粒。cTAG序列可以包括一或多个克隆位点，所述一或多个克隆位点包含但不限于Cas9、Cpf1、限制性和/或重组位点。图2C—一旦整合到生物体的基因组中，cTAG就可以继续用作预先验证的Cas9或Cpf1着陆位点，从而使能够用预先验证的和正交的gRNA序列置换、插入或去除基因组整合的DNA。
图3A-D展示了利用本公开的模块化CRISPR构建体的本发明克隆方法的实施例。图3A—图解了根据本公开的可以用Cas9或Cpf1核酸酶容易地改变的模块化CRISPR质粒。由数字表示的可互换部分侧接有由字母表示的不变的cTAG序列。各部分可以预先组装或者可以基于cTAG序列同一性在体内或在体外组装。示例插入部分示出在图3A的右侧。图3B—可以使用如cTAG处的Cas9、Cpf1或限制性核酸内切酶切割等几种策略来置换单独部分而不必重新组装整个质粒。cTAG序列可以包括一或多个克隆位点，所述一或多个克隆位点包含但不限于Cas9、Cpf1、限制性和/或重组位点。图3C—展示了本公开的用于从现有的模块化质粒中去除插入部分或添加填充物序列的方法。图3D—本公开的模块化质粒的插入部分可以用作用于将模块化CRISPR载体的一部分或全部基因组整合到宿主细胞的基因组中的序列。
图4展示了实例1的一锅式(one-pot)体外模块化CRISPR克隆。具体地，示出了在一锅式反应中通过插入物从一种质粒转移到另一种质粒来生成质粒13001009086。实例1中阐述了此反应的细节。
图5展示了实例2的体外模块化CRISPR克隆方法的实施例。每个图提供了实例2中描述的实验设计的图示。将氯霉素(chloramphenicol)抗性基因克隆到抗卡那霉素(kanamycin)主链质粒中以产生双抗性质粒。然后将双抗性质粒转化为细菌，随后将所述细菌在用卡那霉素和氯霉素抗生素增强的培养基中培养。抗性菌落指示成功的Cpf1克隆组装。
图6展示了实例2的体外模块化CRISPR克隆方法的结果。y轴表示在用卡那霉素和氯霉素增强的培养基中生长的所回收菌落的数量。抗性菌落指示成功的Cpf1克隆组装。结果示出了双抗性质粒的连接酶依赖性组装。
图7描绘了pJDI427的载体图。Cpf1组装中使用的CRISPR着陆位点标记为cTAG M和cTAG N。相关序列信息可以在SEQ ID NO:102中找到。
图8描绘了pJDI429的载体图。Cpf1组装中使用的CRISPR着陆位点标记为cTAG N和cTAG O。相关序列信息可以在SEQ ID NO:103中找到。
图9描绘了pJDI430的载体图。Cpf1组装中使用的CRISPR着陆位点标记为cTAG P和cTAG N。相关序列信息可以在SEQ ID NO:104中找到。
图10描绘了pJDI431的载体图。Cpf1组装中使用的CRISPR着陆位点标记为cTAG P和cTAG O。相关序列信息可以在SEQ ID NO:105中找到。
图11描绘了pJDI432的载体图。Cpf1组装中使用的CRISPR着陆位点标记为cTAG M和cTAG N。相关序列信息可以在SEQ ID NO:106中找到。
图12描绘了pJDI434的载体图。Cpf1C组装中使用的CRISPR着陆位点标记为cTAG N和cTAG O。相关序列信息可以在SEQ ID NO:107中找到。
图13描绘了pJDI435的载体图。Cpf1组装中使用的CRISPR着陆位点标记为cTAG P和cTAG N。相关序列信息可以在SEQ ID NO:108中找到。
图14描绘了pJDI436的载体图。Cpf1组装中使用的CRISPR着陆位点标记为cTAG P和cTAG O。相关序列信息可以在SEQ ID NO:109中找到。
图15展示了根据本公开的方法的对模块化CRISPR构建体的示例基因编辑。具体地，图15展示了使用实例3的大模块化设计通过限制性酶消化和连接的质粒组装。图15示出了模块化CRISPR质粒主链p1300283391和相容的含有GFP的插入DNA部分各自用ApaI和PvuI限制性酶来消化以产生相容的克隆标签末端。消化的主链和插入物在体外连接以产生新的模块化CRISPR构建体。
图16是展示了dCas9与DNA的gRNA定向结合的图。
图17A-B描绘了如实例5中所述在谷氨酸棒状杆菌(Corynebacteriumglutamicum)中的CRISPRi技术验证。图17A—是导致靶基因的转录灭活的dCas9表达载体的图。图17B—描绘了使用与在WT或辣椒(paprika)产生性菌株中表达的各种引导RNA同时表达的dCas9的基因转录阻遏。示出了针对7个生物复制物的活门控细胞的中值荧光。
图18A-B展示了CRISPRi/CRISPRa文库的潜力。图18A—描绘了靶向单个启动子的CRISPRi/CRISPRa文库。图18B—描绘了靶向整个基因组中的多个启动子的CRISPRi/CRISPRa文库。
具体实施方式
定义
虽然认为以下术语可以很好地为本领域的普通技术人员所理解，但是阐述以下定义是为了便于解释当前公开的主题。
术语“一个(a)”或“一种(an)”是指一或多个所述实体，即可以指复数指示物。因此，术语“一个”或“一种”、“一或多个”和“至少一个”在本文可互换地使用。另外，通过不定冠词“一个”或“一种”提及“要素”并不排除存在多于一个要素的可能性，除非上下文明确地要求存在要素中的一个且仅一个要素。
术语“原核生物”是本领域公认的并且是指不含有细胞核的生物体。通常将原核生物分类到细菌和古细菌两个领域之一中。古细菌领域和细菌领域的生物体之间的确切差异是基于16S核糖体RNA中的核苷酸碱基序列的基本差异。
“真核生物”是其细胞含有细胞核和封闭在膜内的其它细胞器的任何生物体。真核生物属于真核域(Eukarya)或真核生物域(Eukaryota)分类单元。将真核细胞与原核细胞(上述细菌和古细菌)区分开的定义性特征是真核细胞具有膜结合的细胞器，特别是细胞核，所述细胞核含有遗传物质并且由核膜封闭。
术语“古细菌”是指疵壁菌(Mendosicute)门生物体的分类、通常发现于不寻常的环境中并且通过若干个标准与其余原核生物区分开，所述若干个标准包含核糖体蛋白的数量和细胞壁中胞壁酸的缺乏。在ssrRNA分析的基础上，古细菌由以下两个系统发育上不同的群组组成：泉古菌(Crenarchaeota)和广古菌(Euryarchaeota)。在其生理学的基础上，古细菌可以组织成三种类型：产甲烷菌(methanogens)(产生甲烷的原核生物)；极端嗜盐菌(halophiles)(生活在很高浓度的盐(NaCl)中的原核生物)；以及极端(超)嗜热菌(thermophilus)(生活在非常高的温度下的原核生物)。除了将原核生物与细菌(即，细胞壁中没有胞壁质、酯连接的膜脂质等)区分开的统一古细菌特征外，这些原核生物表现出使其适应自己的特定栖息地的独特结构或生化属性。泉古菌主要由超嗜热硫依赖性原核生物组成，并且广古菌含有产甲烷菌和极端嗜盐菌。
“细菌”或“真细菌”是指原核生物体的领域。细菌包含如下至少11种不同的群组：(1)革兰氏阳性(gram+)细菌，其中主要有两个亚门：(1)高G+C菌群(放线菌属(Actinomycetes)、分枝杆菌(Mycobacteria)、微球菌属(Micrococcus)等)；(2)低G+C菌群(芽孢杆菌属(Bacillus)、梭状芽胞杆菌属(Clostridia)、乳酸菌属(Lactobacillus)、葡萄球菌属(Staphylococci)、链球菌属(Streptococci)、支原体属(Mycoplasmas))；(2)变形菌门(Proteobacteria)，例如紫色光合的+非光合的革兰氏阴性细菌(包含最“常见的”革兰氏阴性细菌)；(3)蓝细菌(Cyanobacteria)，例如含氧光能利用菌；(4)螺旋菌(Spirochetes)和相关种；(5)浮霉状菌属(Planctomyces)；(6)拟杆菌属(Bacteroides)、黄杆菌(Flavobacteria)；(7)衣原体属(Chlamydia)；(8)绿色硫细菌；(9)绿色非硫细菌(也为厌氧光能利用菌)；(10)抗放射性微球菌和相关物；(11)热袍菌属(Thermotoga)和栖热腔菌属(Thermosipho)嗜热菌。
术语“经过基因修饰的宿主细胞”、“重组宿主细胞”和“重组菌株”在本文中可互换地使用并且是指已经通过本公开的克隆和转化方法进行基因修饰的宿主细胞。因此，术语包含与其所来源于的天然存在的微生物相比已经进行基因改变、修饰或工程化使得其表现出改变的、经过修饰的或不同的基因型和/或表型(例如，当基因修饰影响微生物的编码核酸序列时)的宿主细胞(例如，细菌、酵母细胞、真菌细胞、CHO、人细胞等)。应当理解，术语不仅指所讨论的特定重组微生物，而且还指此类微生物的后代或潜在后代。
术语“基因工程化”可以指对宿主细胞的基因组的任何操纵(例如，通过核酸的插入或缺失)。
如本文所用，“可选择标志物”是通常在特定条件下允许选择用于含有其的分子(例如，复制子)或细胞的核酸区段。这些标志物可以对如但不限于RNA、肽或蛋白质的产生等活性进行编码或者可以为RNA、肽、蛋白质、无机和有机化合物或组合物等提供结合位点。可选择标志物的实例包含但不限于：(1)对提供针对以其它方式有毒的化合物的抗性的产物(例如，抗生素)进行编码的核酸区段；(2)对受体细胞中以其它方式缺乏的产物(例如，tRNA基因、营养缺陷型标志物)进行编码的核酸区段；(3)对抑制基因产物的活性的产物进行编码的核酸区段；(4)对可以容易地识别的产物(例如，表型标志物如β-半乳糖苷酶、绿色荧光蛋白(GFP)、黄色荧光蛋白(YFP)、青色荧光蛋白(CFP)和细胞表面蛋白)进行编码的核酸区段；(5)对结合以其它方式对细胞生存和/或功能有害的其它产物的产物进行编码的核酸区段；(6)对以其它方式抑制任何核酸区段的活性从而产生可见或可选择表型的核酸(例如，反义寡核苷酸)进行编码的核酸区段；(7)对结合修饰底物的其它产物的产物(例如，限制性核酸内切酶)进行编码的核酸区段；(8)可以用于分离或鉴定期望的分子的核酸区段(例如，特定蛋白质结合位点)；(9)对可以以其它方式无功能的特定核苷酸序列(例如，用于分子亚群的PCR扩增)进行编码的核酸区段；以及(10)当其不存在时直接或间接向特定化合物赋予抗性或敏感性的核酸区段。
如本文所用，“可逆选择标志物”或“逆选择标志物”是在选择时消除或抑制宿主生物体的生长的核酸区段。在一些实施例中，本公开的可逆选择标志物使细胞对一或多种化学品/生长条件/遗传背景敏感。在一些实施例中，本公开的可逆选择标志物是毒性基因。在一些实施例中，可逆选择标志物由诱导型启动子表达。
如本文所用，术语“核酸”是指任何长度的核苷酸的聚合形式核糖核苷酸或脱氧核糖核苷酸或其类似物。此术语指分子的一级结构并且因此包含双链和单链DNA以及双链和单链RNA。所述术语还包含经过修饰的核酸，如甲基化和/或加帽的核酸、含有经过修饰的碱基的核酸、主链修饰等。术语“核酸”和“核苷酸序列”可互换使用。
如本文所用，术语“基因”是指与生物功能相关的DNA的任何区段。因此，基因包含但不限于其表达所需的编码序列和/或调节序列。基因还可以包含例如形成其它蛋白质的识别序列的非表达的DNA区段。基因可以从多种来源获得(包含从感兴趣来源克隆或从已知或预测的序列信息合成)并且可以包含被设计成具有期望的参数的序列。
如本文所用，术语“同源”或“同源物”或“直系同源物”是本领域已知的并且是指共享共同祖先或家族成员并且基于序列同一性程度确定的有关序列。术语“同源性”、“同源”、“基本上类似”和“基本上对应”在本文可互换地使用。所述术语是指其中一或多个核苷酸碱基的改变不影响核酸片段介导基因表达或产生某种表型的能力的核酸片段。这些术语还指本公开的核酸片段的修饰，如相对于初始的未经过修饰的片段基本上不改变所得核酸片段的功能性质的一或多个核苷酸的缺失或插入。因此，如本领域技术人员将理解的，应当理解，本公开不仅仅涵盖具体的示范性序列。这些术语描述在一种物种、亚种、变种、栽培品种或菌株中发现的基因与另一物种、亚种、变种、栽培品种或菌株中的对应或等效基因之间的关系。出于本公开的目的，对同源序列进行比较。认为、相信或已知“同源序列”或“同源物”或“直系同源物”在功能上相关。可以通过多种方式中的任何一种方式指示功能关系，包含但不限于：(a)序列同一性程度和/或(b)相同或类似的生物功能。优选地，指示(a)和(b)两者。同源性可以使用本领域中容易获得的软件程序来确定，如在以下中讨论的那些：当代分子生物学实验指南(Current Protocols in Molecular Biology)(F.M.奥苏贝尔(F.M.Ausubel)等人编,1987)增刊30,第7.718章,表7.71。一些比对程序是MacVector(英国牛津的牛津分子有限公司(Oxford Molecular Ltd,Oxford,U.K.))、ALIGN Plus(宾夕法尼亚州的科学与教育软件公司(Scientific and Educational Software,Pennsylvania))和AlignX(载体NTI(Vector NTI)，加利福尼亚州卡尔斯巴德市的英杰生命技术有限公司(Invitrogen,Carlsbad,CA))。另一个比对程序是使用默认参数的Sequencher(密歇根州安阿伯市的基因编码公司(Gene Codes,Ann Arbor,Michigan))。
如本文所用，如本领域所熟知的，术语“核苷酸变化”是指例如核苷酸取代、缺失和/或插入。例如，突变含有产生沉默取代、添加或缺失但不改变编码蛋白质的性质或活性或蛋白质的制备方式的改变。
如本文所用，如本领域所熟知的，术语“蛋白质修饰”是指例如氨基酸取代、氨基酸修饰、缺失和/或插入。
如本文所用，术语核酸或多肽的“至少一部分”或“片段”意指具有此类序列的最小尺寸特性的部分或全长分子的至多并包含全长分子的任何较大片段。本公开的多核苷酸的片段可以对基因调节元件的生物活性部分进行编码。基因调节元件的生物活性部分可以通过分离本公开的多核苷酸之一的包括基因调节元件的部分并且如本文所描述的对活性进行评估来制备。类似地，多肽的一部分可以是4个氨基酸、5个氨基酸、6个氨基酸、7个氨基酸，依此类推，直至全长多肽。待使用的部分的长度将取决于特定的应用。核酸的可用作杂交探针的部分可以短至12个核苷酸；在一些实施例中，所述部分为20个核苷酸。多肽的可用作表位的部分可以短至4个氨基酸。多肽的执行全长多肽的功能的部分通常长于4个氨基酸。
对于本文中公开的多核苷酸的PCR扩增，可以将寡核苷酸引物设计成用于在PCR反应中使用以从提取自任何感兴趣生物体的cDNA或基因组DNA扩增对应的DNA序列。用于设计PCR引物和PCR克隆的方法在本领域中通常是已知的并且在以下中公开：萨姆布鲁克(Sambrook)等人(2001)分子克隆：实验室手册(Molecular Cloning:A LaboratoryManual)(第3版,纽约普莱恩维尤的冷泉港实验室出版社(Cold Spring HarborLaboratory Press,Plainview,New York))。还参见英尼斯(Innis)等人编.(1990)PCR方案：方法和应用指南(PCR Protocols:A Guide to Methods and Applications)(纽约学术出版社(Academic Press,New York))；英尼斯和盖尔芬德(Gelfand)编.(1995)PCR策略(PCR Strategies)(纽约学术出版社)；以及英尼斯和盖尔芬德编.(1999)PCR方法手册(PCRMethods Manual)(纽约学术出版社)。PCR的已知方法包含但不限于使用配对引物、嵌套引物、单特异性引物、简并引物、基因特异性引物、载体特异性引物、部分错配引物等的方法。
如本文所用，术语“引物”是指能够与允许DNA聚合酶连接的扩增靶标退火的寡核苷酸，由此当被置于引物延伸产物的合成被诱导的条件下时即在核苷酸和如DNA聚合酶等聚合剂的存在下以及在适当的温度和pH下用作DNA合成的起始点。(扩增)引物优选地是单链的以得到最大扩增效率。优选地，引物是寡脱氧核糖核苷酸。引物必须足够长以在聚合剂的存在下引发延伸产物的合成。引物的确切长度将取决于许多因素，包含温度和引物的组成(A/T对G/C含量)。一对双向引物由一个正向引物和一个反向引物组成，如DNA扩增领域中如PCR扩增中常用的。
术语“严格度”或“严格杂交条件”是指影响杂交体的稳定性的杂交条件，例如温度、盐浓度、pH、甲酰胺浓度等。根据经验优化这些条件，以使引物或探针与其靶核酸序列的特异性结合最大化并使其非特异性结合最小化。所用的术语包含指探针或引物将与其靶序列杂交至比其它序列可检测地更大的程度(例如，至少2倍于背景)的条件。严格条件是与取决于序列的并且将在不同的情形下有所不同。较长的序列在较高的温度下特异性地杂交。通常，严格条件被选择为在限定的离子强度和pH下比特异性序列的热熔点(Tm)低约5℃。Tm是(在限定的离子强度和pH下)50％的互补靶序列与完全匹配的探针或引物杂交时的温度。严格条件通常将是这样的条件：其中在pH 7.0至8.3下盐浓度小于约1.0M Na+离子，通常为约0.01至1.0M Na+离子浓度(或其它盐)，并且对于短探针或引物(例如，10至50个核苷酸)，温度为至少约30℃，而对于长探针或引物(例如，大于50个核苷酸)，温度至少约60℃。严格条件还可以通过添加如甲酰胺等去稳定剂来实现。示范性低严格条件或“降低的严格度的条件”包含与30％甲酰胺、1M NaCl、1％SDS的缓冲溶液在37℃下杂交并且在2×SSC中在40℃下洗涤。示范性高严格度条件包含在50％甲酰胺、1M NaCl、1％SDS中在37℃下杂交并且在0.1×SSC中在60℃下的洗涤。杂交程序是本领域熟知的并且由例如奥苏贝尔等人,1998和萨姆布鲁克等人,2001描述。在一些实施例中，严格条件是在45℃下在含有1mM Na2EDTA、0.5-20％十二烷基硫酸钠的0.25M Na2HPO4缓冲液(pH 7.2)中杂交，如0.5％、1％、2％、3％、4％、5％、6％、7％、8％、9％、10％、11％、12％、13％、14％、15％、16％、17％、18％、19％或20％，然后在55℃到65℃下在含有0.1％(w/v)十二烷基硫酸钠的5×SSC中洗涤。
如本文所用，术语“基本相同”是指两个多核苷酸序列的差异不多于1个、2个、3个、4个、5个、6个或7个核苷酸。当在cTAG的上下文中使用时，术语基本相同表示除了两个cTAG之一上的被设计成消除至少一个CRISPR着陆位点中的CRISPR切割的在PAM或原间隔子区域中的突变之外，所述cTAG将相同。当术语基本相同与术语“部分”序列或cTAG结合使用时，所述组合是指如上所述的两个基本相同cTAG之间的比较，其中cTAG之一已被CRISPR核酸内切酶消化。因此，术语将用于指示所描述的cTAG除了在PAM或原间隔子区域中的突变之外与第二cTAG(呈其未被消化的形式)相同。
如本文所用，术语“启动子”是指能够控制编码序列或功能性RNA的表达的DNA序列。启动子序列可以由近端和更远端的上游元件组成，后一种元件通常被称为增强子。因此，“增强子”是可以刺激启动子活性的DNA序列并且可以是启动子的先天元件或被插入以增强启动子的水平或组织特异性的异源元件。
如本文所用，术语“异源”是指未在特定生物体中天然发现的核酸序列。
如本文所用，术语“内源”、“内源基因”是指基因的天然存在的拷贝。
如本文所用，术语“天然存在的”是指来源于天然存在的来源的基因或序列。在一些实施例中，天然存在的基因是指野生型(非转基因)基因的基因，无论当被引入不同生物体中时定位于其源生物体内的内源环境中还是置于“异源”环境中。因此，出于本公开的目的，“非天然存在的”序列是已合成、突变或以其它方式修饰以具有与已知天然序列不同的序列的序列。在一些实施例中，修饰可以处于蛋白质水平下(例如，氨基酸取代)。在其它实施例中，修饰可以处于DNA水平下，而对蛋白质序列没有任何影响(例如，密码子优化)。在一些实施例中，非天然存在的序列可以是构建体。
如本文所用，术语“外源”与术语“异源”可互换地使用并且是指来自除其天然来源之外的某个来源的物质。例如，术语“外源蛋白”或“外源基因”是指来自非天然来源或定位并且已被人工供应给生物系统的蛋白质或基因。出于本公开的目的，内源基因的人工突变的变体被认为是“外源的”。
如本文所用，短语“重组构建体”、“表达构建体”、“嵌合构建体”、“构建体”和“重组DNA构建体”在本文可互换地使用。重组构建体包括核酸片段的人工组合，例如自然界中未一起发现的调节序列和编码序列。例如，嵌合构建体可以包括源自不同来源的调节序列和编码序列或源自相同来源但以与自然界中发现的方式不同的方式布置的调节序列和编码序列。此类构建体可以单独使用或者可以与载体结合使用。如果使用载体，则载体的选择取决于如本领域技术人员所熟知的将用于转化宿主细胞的方法。例如，可以使用质粒载体。技术人员充分了解为了成功转化、选择和繁殖包括本公开的分离的核酸片段中的任何分离的核酸片段的宿主细胞而必须存在于载体上的基因元件。技术人员还将认识到，不同的独立转化事件将导致不同的表达水平和模式(琼斯(Jones)等人,(1985)欧洲分子生物学组织期刊(EMBO J.)4:2411-2418；阿尔梅达(De Almeida)等人,(1989)分子遗传学与基因组学(Mol.Gen.Genetics)218:78-86)，并且因此，必须筛选多个事件以获得显示期望的表达水平和模式的线。可以通过DNA的Southern分析、mRNA表达的Northern分析、蛋白表达的免疫印迹分析或表型分析等来完成此类筛选。载体可以是自主复制或可以整合到宿主细胞的染色体中的质粒、病毒、噬菌体、前病毒、噬菌粒、转座子、人工染色体等。载体也可以是不自主复制的裸RNA多核苷酸、裸DNA多核苷酸、由同一链内的DNA和RNA两者构成的多核苷酸、聚赖氨酸缀合的DNA或RNA、肽缀合的DNA或RNA、脂质体缀合的DNA等。如本文所用，术语“表达”是指功能性最终产物例如mRNA或蛋白质(前体或成熟)的产生。
术语“可操作地连接”意指在上下文中根据本公开的启动子多核苷酸与另外一个寡核苷酸或多核苷酸的顺序布置，从而导致所述另外一个多核苷酸的转录。在一些实施例中，本公开的启动子序列恰好插入在基因的5'UTR或开放阅读框之前。在其它实施例中，本公开的可操作地连接的启动子序列和基因序列被一或多个接头核苷酸分开。在CRISPR原间隔子和原间隔子邻近基序(PAM)的上下文中，术语“可操作地连接”是指能够被CRISPR核酸内切酶复合物高效切割的靠近放置的原间隔子/PAM组合序列。在引导RNA/适配子的上下文中，术语“可操作地连接”是指能够将CRISPR核酸内切酶募集到DNA靶位点，同时还通过其适配子序列募集第二效应子肽(例如，能够募集适配子所靶向的转录激活结构域)的引导RNA。在终止子序列的上下文中，术语“可操作地连接”意指终止子序列的用于终止上游序列的转录的布置。在一些实施例中，终止子序列放置在基因或操纵子的末端。
术语“CRISPR RNA”或“crRNA”是指负责与靶DNA序列杂交并募集CRISPR核酸内切酶的RNA链。crRNA可以是天然存在的或者可以根据产生RNA的任何已知方法合成。
术语“引导序列”或“间隔子”是指crRNA或引导RNA(gRNA)中的负责与靶DNA杂交的部分。
术语“原间隔子”是指由crRNA或引导链靶向的DNA序列。在一些实施例中，原间隔子序列与CRISPR复合物的crRNA引导序列杂交。
术语“种子区域”是指负责DNA序列CRISPR核糖核蛋白复合物之间的初始复合的核糖核酸序列。与crRNA/sgRNA序列的剩余部分相比，种子区域与靶DNA序列之间的错配对靶位点识别和切割的影响更强。在一些实施例中，crRNA/gRNA的种子区域中的单个错配可以使CRISPR复合物在所述结合位点处无活性。在一些实施例中，Cas9核酸内切酶的种子区域沿着引导序列的3'部分的最后约12个nt定位，所述最后约12个nt对应于原间隔子靶序列的与PAM相邻的部分(与其杂交)。在一些实施例中，Cpf1核酸内切酶的种子区域沿着引导序列的5'部分的约前5个nt定位，所述约前5个nt对应于原间隔子靶序列的与PAM相邻的部分(与其杂交)。
在CRISPR的上下文中，术语“靶位点”是指引导RNA(例如，单引导RNA或tracrRNA)与其对应的种子区域复合使得引导RNA将能够将CRISPR核酸内切酶(活性的或另外的)募集到DNA的所述部分的基因座。
术语“tracrRNA”是指小的反式编码的RNA。tracrRNA与crRNA互补并与crRNA碱基配对，以形成能够将CRISPR核酸内切酶募集到靶序列的crRNA/tracrRNA杂交体。
如本文所用，术语“引导RNA”或“gRNA”是指能够将CRISPR核酸内切酶募集到靶序列的RNA序列或序列的组合。因此，如本文所用，引导RNA可以是天然或合成的crRNA(例如，对于Cpf1)、天然或合成的crRNA/tracrRNA杂交体(例如，对于Cas9)或单引导RNA(sgRNA)。因此，叙述针对引导RNA的表达或核苷酸编码的权利要求可以指将单引导RNA序列、crRNA和/或crRNA和tracrRNA两者表达/编码为不同分子。
如本文所用，术语“CRISPR着陆位点”是指能够由CRISPR复合物靶向的DNA序列。因此，在一些实施例中，CRISPR着陆位点包括能够由CRISPR核酸内切酶复合物切割的靠近放置的原间隔子/原间隔子邻近基序组合序列。术语“经过验证的CRISPR着陆位点”是指存在能够诱导所述序列的高效切割的引导RNA的CRISPR着陆位点。因此，术语经过验证的应被解释为意指序列先前已经被示出可由CRISPR复合物切割。每个“经过验证的CRISPR着陆位点”将根据定义确认与验证相关联的经过测试的引导RNA的存在。术语“经过验证的CRISPR着陆位点”应进一步被理解为意指着陆位点被人工设计并添加到DNA序列中，其明确目的是用作高效且可靠的DNA切割靶标。因此，本公开的“经过验证的CRISPR着陆位点”排除先前存在的质粒中的序列，所述序列最初未被设计为CRISPR靶向位点，但是随后通过质粒的定制CRISPR复合物靶向区域的产生而被切割。
术语“一或多个粘性末端”是指包括序列突出端的双链多核苷酸分子末端。在一些实施例中，粘性末端可以是具有5'或3'序列突出端的dsDNA分子末端。在一些实施例中，本公开的粘性末端能够与相同或其它分子的相容的粘性末端杂交。因此，在一个实施例中，第一DNA片段的3'上的粘性末端可以与第二DNA片段上的相容的粘性末端杂交。在一些实施例中，这些杂交的粘性末端可以通过连接酶缝合在一起。在其它实施例中，粘性末端可能需要突出端的延伸以在连接之前完成dsDNA分子。术语“一或多个基因瘢痕”是指通过DNA操纵方法引入核酸序列中的任何不期望的序列。例如，在一些实施例中，本公开教导了基因瘢痕，如限制性酶结合位点、用于适应克隆的序列适配子或间隔子、TA位点、从NHEJ中遗留下来的瘢痕等。在一些实施例中，本公开教导了无瘢痕克隆和基因编辑的方法。
如本文所用，术语“靶向的”是指一个项或分子将与另一个项或分子以一定程度的特异性相互作用以排除非靶向的项或分子的预期。例如，已经将根据本公开的靶向至第二多核苷酸的第一多核苷酸设计成以序列特异性方式(例如，通过沃森-克里克(Watson-Crick)碱基配对)与第二多核苷酸杂交。在一些实施例中，设计了杂交的选择区域以便使杂交对于一或多个靶向的区域是独特的。如果第二多核苷酸的靶向序列(杂交区域)发生突变或者以其它方式从第二多核苷酸中去除/分离，则第二多核苷酸可以不再是第一靶向多核苷酸的靶标。
本公开将所教导和描述的通用模块化CRISPR DNA构建体或设计称作“大模块化”构建体或设计。
DNA核酸酶
在一些实施例中，本公开教导了用于利用DNA核酸酶进行基因编辑/克隆的方法和组合物。CRISPR复合物、转录激活因子样效应子核酸酶(TALEN)、锌指核酸酶(ZFN)和FokI限制性酶是已被用作基因编辑工具的一些序列特异性核酸酶。这些酶能够通过与被工程化以识别感兴趣序列的引导区域相互作用来将其核酸酶活性靶向期望的靶基因座。在一些实施例中，本公开教导了基于CRISPR的基因编辑方法
基于CRISPR的体内编辑的原理在很大程度上依赖于天然细胞DNA修复系统。由核酸酶引入的双链dsDNA断裂通过非同源末端连接(NHEJ)或同源性定向修复(HDR)或单链退火(SSA)或微同源性末端连接(MMEJ)进行修复。
HDR依赖于含有与靶向的DNA切割位点周围的区域同源的序列的模板DNA。细胞修复蛋白使用外源性地供应的或内源DNA序列与DNA断裂周围的位点之间的同源性来修复dsDNA断裂，从而用模板DNA上的序列来置换断裂。但是，整合模板DNA失败可能会导致NHEJ、MMEJ或SSA。NHEJ、MMEJ和SSA是易错过程，所述易错过程通常伴随靶位点处的核苷酸插入或缺失(插入缺失)，从而由于提前终止密码子的移码突变或插入而导致基因组的靶向的区域的基因敲除(沉默)。Cpf1介导的编辑也可以通过由核酸内切酶产生的突出端的传统杂交、然后进行连接来起作用。
CRISPR核酸内切酶还可用于体外DNA操纵，如本公开的稍后部分所讨论的。
CRISPR系统
CRISPR(成簇规律间隔短回文重复序列)和CRISPR关联的(cas)核酸内切酶最初被发现是由细菌和古细菌进化的防止病毒和质粒入侵的适应性免疫系统。细菌中天然存在的CRISPR/Cas系统由一或多个Cas基因和一或多个CRISPR阵列组成，所述一或多个CRISPR阵列由碱基序列的短回文重复序列组成，所述碱基序列由从先前遇到的病毒和质粒(称为间隔子)获取的基因组靶向序列分开。(维登海夫特(Wiedenheft),B.等人自然(Nature).2012；482:331；巴哈亚(Bhaya),D.等人,遗传学年评(Annu.Rev.Genet.)2011；45:231；以及特尔姆斯(Terms),M.P.等人,微生物学当前观点(Curr.Opin.Microbiol.)2011；14:321)。具有一或多个CRISPR基因座的细菌和古细菌通过将外来序列的短片段(原间隔子)在CRISPR阵列的近端处整合到宿主染色体中来响应病毒或质粒挑战。CRISPR基因座的转录生成了含有与先前遇到的入侵核酸互补的序列的CRISPR源性RNA(crRNA)文库(豪维兹(Haurwitz),R.E.等人,科学(Science).2012:329；1355；格斯纳(Gesner),E.M.等人,自然结构和分子生物学(Nat.Struct.Mol.Biol.)2001:18；688；季聂克(Jinek),M.等人,科学.2012:337；816-21)。由crRNA进行的靶识别是通过与靶DNA的互补碱基配对发生的，所述靶DNA通过Cas蛋白指导外来序列的切割。(季聂克等人2012“适应性细菌免疫中的可编程双RNA引导DNA核酸内切酶(A Programmable dual-RNA-guided DNA endonuclease inadaptive bacterial immunity)”科学.2012:337；816-821)。
存在至少五种主要的CRISPR系统类型(I型、II型、III型、IV型和V型)和至少16种不同的子类型(马卡洛娃(Makarova),K.S.等人,自然综述：微生物学(Nat RevMicrobiol.)2015.自然综述：微生物学13,722-736)。CRISPR系统也基于其效应蛋白进行分类。1类系统具有多亚基crRNA-效应子复合物，而在2类系统中，效应子复合物的所有功能均由单个蛋白质(例如，Cas9或Cpf1)进行。在一些实施例中，本公开教导了使用II型和/或V型单亚基效应子系统。因此，在一些实施例中，本公开教导了使用2类CRISPR系统。
CRISPR/Cas9
在一些实施例中，本公开教导了使用II型CRISPR系统进行基因编辑的方法。在一些实施例中，II型CRISPR系统使用Cas9酶。II型系统依赖于i)单个核酸内切酶蛋白、ii)反式激活的crRNA(tracrRNA)和iii)crRNA，其中crRNA的5'端的约20个核苷酸(nt)的部分与靶核酸互补。CRISPR crRNA链的与其靶DNA原间隔子互补的区域在此称为“引导序列”。
在一些实施例中，II型系统的tracrRNA和crRNA组分可以被单引导RNA(sgRNA)置换。sgRNA可以包含例如包括与靶DNA序列(引导序列)互补的至少12-20个核苷酸的序列的核苷酸序列并且可以在其3'端处包含共同的支架RNA序列。如本文所用，“共同的支架RNA”是指模拟tracrRNA序列的任何RNA序列或用作tracrRNA的任何RNA序列。
Cas9核酸内切酶产生平端DNA断裂并通过crRNA和tracrRNA寡核苷酸的组合募集到靶DNA，所述tracrRNA寡核苷酸通过RNA CRISPR复合物的互补杂交来拴系核酸内切酶。(参见图1A中的实心三角形箭头)。
在一些实施例中，由crRNA/核酸内切酶复合物进行的DNA识别需要与位于靶原间隔子下游的靶DNA的3'部分的原间隔子邻近基序(PAM)(例如，5'-NGG-3')进行另外的互补碱基配对。(季聂克,M.等人,科学.2012:337；816-821)。在一些实施例中，由Cas9识别的PAM基序对于不同的Cas9蛋白质是不同的。
在一些实施例中，本领域技术人员可以理解，本文中公开的Cas9可以是源自或分离自任何来源的任何变体。例如，在一些实施例中，本公开的Cas9肽可以包含选自SEQ IDNO:1、SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5和SEQ ID NO:6的SEQ ID No中的一或多个。在其它实施例中，本公开的Cas9肽可以包含文献中所描述的突变中的一或多个突变，包含但不限于以下中所描述的功能性突变：方法拉(Fonfara)等人,核酸研究(Nucleic Acids Res.)2014年2月；42(4):2577-90；西村H.(Nishimasu H.)等人细胞(Cell).2014年2月27日；156(5):935-49；季聂克M.等人科学.2012 337:816-21；以及季聂克M.等人科学.2014年3月14日；343(6176)；还参见2013年3月15日提交的美国专利申请第13/842,859号，所述美国专利申请通过引用的方式并入在此；进一步地，参见美国专利第8,697,359号；8,771,945；8,795,965；8,865,406；8,871,445；8,889,356；8,895,308；8,906,616；8,932,814；8,945,839；8,993,233；以及第8,999,641号，所述美国专利全部通过引用的方式并入在此。因此，在一些实施例中，本文所公开的系统和方法可以与具有双链核酸酶活性的野生型Cas9蛋白、充当单链切口酶的Cas9突变体或具有经过修饰的核酸酶活性的其它突变体一起使用。
如在本文件的稍后部分中进一步详细地描述的，本公开进一步设想了催化灭活Cas9突变体的用途。在一些实施例中，术语“催化灭活”或“无催化活性”CRISPR是指其中DNA酶催化结构域无功能(即，酶不再切割DNA)的CRISPR蛋白。因此，在一些实施例中，本公开教导了dCas9突变体。减少或消除Cas9中的核酸酶的突变的非限制性列表包含：D10、G12、G17、E762、H840、N854、N863、H982、H983、A984、D986或A987或在Cas9同源物或直系同源物中的对应定位的突变。一或多个突变可以包含被任何天然(例如,丙氨酸)或非天然氨基酸取代或缺失。示范性核酸酶缺陷dCas9蛋白是Cas9D10A&H840A(季聂克等人,科学.2012年8月17日；337(6096):816-21；齐(Qi)等人,细胞.2013年2月28日；152(5):1173-83)。表1中提供了dCas9变体的非限制性列表。
表1：dCas9载体的非限制性列表
Figure BDA0002979126820000231
Figure BDA0002979126820000241
Figure BDA0002979126820000251
Figure BDA0002979126820000261
Figure BDA0002979126820000271
Figure BDA0002979126820000281
Figure BDA0002979126820000291
CRISPR/Cpf1
在其它实施例中，本公开教导了使用V型CRISPR系统进行基因编辑的方法。在一些实施例中，本公开教导了使用来自普氏菌属(Prevotella)和弗朗西斯氏菌属1(Francisella 1)(Cpf1)的CRISPR的方法。
本公开的Cpf1 CRISPR系统包括i)单个核酸内切酶蛋白和ii)crRNA，其中crRNA的3'端的一部分含有与靶核酸互补的引导序列。在此系统中，Cpf1核酸酶通过crRNA直接募集到靶DNA(参见图1B中的实心三角箭头)。在一些实施例中，Cpf1的引导序列必须是至少12nt、13nt、14nt、15nt或16nt以实现可检测的DNA切割并且最少14nt、15nt、16nt、17nt或18nt以实现高效的DNA切割。
本公开的Cpf1系统以各种方式中不同于Cas9。第一，与Cas9不同，Cpf1不需要单独的tracrRNA用于切割。在一些实施例中，Cpf1 crRNA可以短至约42-44个碱基长，其中23-25nt是引导序列，并且19nt是组成型直接重复序列。相比之下，组合的Cas9tracrRNA和crRNA合成序列可以为约100个碱基长。在一些实施例中，本公开将会将用于Cpf1的crRNA称为“引导RNA”。
第二，Cpf1优选位于在其靶标上游的5'的“TTN”PAM基序。这与位于针对Cas9系统的靶DNA的3'上的“NGG”PAM基序形成对照。在一些实施例中，紧接在引导序列之前的尿嘧啶碱基不能被取代(蔡澈(Zetsche),B.等人2015.“Cpf1是2类CRISPR-Cas系统的单一RNA引导的核酸内切酶(Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-CasSystem)”细胞163,759-771，所述文献通过全文引用的方式并入在此)。
第三，针对Cpf1的切割位点错开了约3-5个碱基，所述碱基形成了“粘性末端”(金姆(Kim)等人,2016.“全基因组分析揭示了人细胞中Cpf1核酸内切酶的特异性(Genome-wide analysis reveals specificities of Cpf1 endonucleases in human cells)”2016年06月在线发表)。具有约3-5nt突出端的这些粘性末端被认为促进NHEJ介导的连接并改善具有匹配末端的DNA片段的基因编辑。切割位点在靶DNA的3'端，所述端在PAM所在的5'端的远端。切割位置通常在非杂交链上的第18个碱基和与crRNA杂交的互补链上的第23个碱基之后(图1B)。
第四，在Cpf1复合物中，“种子”区域位于引导序列的前5nt之内。Cpf1 crRNA种子区域对突变高度敏感，并且即使所述区域中的单碱基取代也可以大大降低切割活性(参见蔡澈,B.等人2015“Cpf1是2类CRISPR-Cas系统的单一RNA引导的核酸内切酶”细胞163,759-771)。至关重要的是，与Cas9 CRISPR靶标不同，Cpf1系统的切割位点和种子区域不重叠。可按照以下获得关于设计靶向Cpf1 crRNA的寡核苷酸的另外指南：(蔡澈,B.等人2015.“Cpf1是2类CRISPR-Cas系统的单一RNA引导的核酸内切酶”细胞163,759-771)。
本领域技术人员将理解，本文中公开的Cpf1可以是源自或分离自任何来源的任何变体。例如，在一些实施例中，本公开的Cpf1肽可以包含选自以下的SEQ ID No中的一或多个：SEQ ID NO:7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50、51、52、53、54、55、56、57、58、59、60、61、62、63、64或其任何变体。
如在本文的稍后部分中进一步详细地描述的，本公开进一步设想了催化灭活Cpf1突变体的用途。因此，在一些实施例中，本公开教示Cpf1突变体。在一些实施例中，本公开的Cpf1包括：ddCpf1(张(Zhang)等人“通过CRISPR ddCpf1进行的多重基因调节(Multiplexgene regulation by CRISPR ddCpf1)”细胞发现(Cell Discovery),3,文章编号17018(2017))；新凶手弗朗西丝菌(Francisella novicida)(UniProtKB—A0Q7Q2(CPF1_FRATN))、毛螺菌科细菌(Lachnospiraceae bacterium)(UniProtKB—A0A182DWE3(A0A182DWE3_9FIRM))和氨基酸球菌(Acidaminococcus sp.)(UniProtKB—U2UMQ6(CPF1ACISB)。在一些实施例中，本公开的dCpf1是通过使催化结构域AsCpf1发生突变而产生的(D908A，山野(Yamano),T.,西村(Nishimasu),H.,蔡澈,B.,平野(Hirano),H.,史雷梅克(Slaymaker),I.M.,李(Li),Y.,费德洛娃(Fedorova),I.,中根(Nakane),T.,马卡洛夫(Makarova),K.S.,库宁(Koonin),E.V.等人(2016)与引导RNA和靶DNA复合的Cpf1的晶体结构(Crystal Structure of Cpf1 in Complex with Guide RNA and Target DNA).细胞,165,949-962)。
连接酶
在一些实施例中，本公开教导了通过靶向的Cpf1复合物来切割靶DNA并且然后将所得的粘性末端与DNA插入物连接的方法。在一些实施例中，本公开教导了提供用于切割靶DNA的Cpf1复合物和用于将DNA“缝合”回到一起的连接酶的方法。在其它实施例中，本公开教导了包含拴系的连接酶的经过修饰的Cpf1复合物。
如本文所用，术语“连接酶”可以包括任何数量的酶促或非酶促试剂。例如，连接酶是酶促连接试剂或催化剂，所述酶促连接试剂或催化剂在适当条件下在DNA分子、RNA分子或杂交体中的邻近核苷酸的3'-OH与5'-磷酸之间形成磷酸二酯键。
在一些实施例中，本公开教导了酶促连接酶的用途。相容的温度敏感性酶促连接酶包含但不限于噬菌体T4连接酶和大肠杆菌连接酶。热稳定连接酶包含但不限于Afu连接酶、Taq连接酶、Tfl连接酶、Tth连接酶、Tth HB8连接酶、栖热菌(Thermus species)AK16D连接酶和Pfu连接酶(参见例如公开的P.C.T.申请WO/2000/026381；吴(Wu)等人,基因(Gene),76(2):245-254,(1989)；以及罗(Luo)等人,核酸研究,24(15):3071-3078(1996))。本领域技术人员将理解，可以从嗜热或超嗜热生物体例如某些真细菌和古细菌物种获得任何数量的热稳定连接酶；而且，此类连接酶可以用于公开的方法和试剂盒中。在一些实施例中，可逆灭活酶(参见例如美国专利第5,773,258号)可以用于本发明教导的一些实施例中。
在其它实施例中，本公开教导了化学连接剂的用途。化学连接剂包含但不限于活化剂、缩合剂和还原剂，如碳二亚胺、溴化氰(BrCN)、N-氰基咪唑、咪唑、1-甲基咪唑/碳二亚胺/胱胺、二硫苏糖醇(DTT)和紫外线。自动连接即在不存在连接剂的情况下的自发连接也在本文的教导的范围内。化学连接方法的详细方案和适当的反应性基团的描述可以在以下以及其它地方找到：许(Xu)等人,核酸研究,27:875-81(1999)；格里亚兹诺夫(Gryaznov)和莱辛格(Letsinger),核酸研究21:1403-08(1993)；格里亚兹诺夫等人,核酸研究22:2366-69(1994)；金谷(Kanaya)和柳川(Yanagawa),生物化学(Biochemistry)25:7423-30(1986)；吕布克(Luebke)和德文(Dervan),核酸研究20:3005-09(1992)；西弗斯(Sievers)和冯·基德罗夫斯基(von Kiedrowski),自然369:221-24(1994)；刘(Liu)和泰勒(Taylor),核酸研究26:3300-04(1999)；王(Wang)和库尔(Kool),核酸研究22:2326-33(1994)；普马尔(Purmal)等人,核酸研究20:3713-19(1992)；阿什利(Ashley)和库什兰(Kushlan),生物化学30:2927-33(1991)；楚(Chu)和奥格尔(Orgel),核酸研究16:3671-91(1988)；索科洛娃(Sokolova)等人,FEBS快报(FEBS Letters)232:153-55(1988)；奈勒(Naylor)和吉勒姆(Gilham),生物化学5:2722-28(1966)；以及美国专利第5,476,930号。
在一些实施例中，本公开的方法、试剂盒和组合物还与光连接反应相容。使用适当波长的光作为连接剂的光连接也在教导的范围内。在一些实施例中，光连接包括探针，所述探针包括核苷酸类似物，所述核苷酸类似物包含但不限于4-硫代胸苷、5-乙烯基尿嘧啶及其衍生物或其组合。在一些实施例中，连接剂包括：(a)在UV-A范围(约320nm到约400nm)内、UV-B范围(约290nm到约320nm)或其组合内的光；(b)波长介于约300nm与约375nm之间的光；(c)波长为约360nm到约370nm的光；(d)波长为约364nm到约368nm的光；或(e)波长为约366nm的光。在一些实施例中，光连接是可逆的。光连接的描述可以在以下以及其它地方找到：藤本(Fujimoto)等人,核酸研讨会丛刊(Nucl.Acid Symp.Ser.)42:39-40(1999)；藤本等人,核酸研讨会丛刊增刊(Nucl.Acid Res.Suppl.)1:185-86(2001)；藤本等人,核酸增刊(Nucl.Acid Suppl.),2:155-56(2002)；刘(Liu)和泰勒(Taylor),核酸研究26:3300-04(1998)以及万维网sbchem.kyoto-u.ac.jp/saito-lab。
通用模块化CRISPR DNA构建体和其用途
在一些实施例中，本发明描述了用于DNA构建体的模块化组装的策略。在一些实施例中，本公开的DNA组装方法适用于任何构建体，包含质粒、小线性DNA和转化的染色体基因座。
在各方面，发明人将此类通用模块化CRISPR DNA构建体称为“大模块化”设计。
传统DNA编辑和组装技术中的缺点
传统的多组分DNA克隆策略在有效组装和修饰具有复杂序列的多组分DNA构建体方面能力有限。例如，限制性酶克隆受到独特限制性酶识别位点的可用性的限制，所述限制性酶识别位点适当地位于DNA插入物中的每个DNA插入物处的克隆接合点以及其在最终载体内的目的地位点处。捷威克隆(gateway cloning)技术类似地受到可用于多组分组装的相对少量的独特重组位点的限制。
传统DNA组装技术的另一个缺点是其编辑序列的能力通常受限于构建时间。例如，如连接酶循环反应(LCR)(一旦初始组装完成，LCR就不容易被修改)等高效组装策略的产物(科克(Kok),S等人,2014“通过连接酶循环反应进行的快速且可靠DNA组装(Rapid andReliable DNAassembly via Ligase Cycling Reaction)”ACS合成生物学(ACSSynth.Biol.),3(2):97-106)。类似的担忧也随着传统的限制性酶克隆出现，一旦含有限制性位点的多核苷酸插入组装的构建体中或者当所述构建体整合到充满所述位点的染色体中时，所述限制性酶克隆的共同限制性识别位点就会停止充当独特的克隆点。因此，一旦克隆过程顺利进行，通过顺序限制性克隆产生的载体就提供非常少的选项用于固定或更新序列。
即使是如传统的CRISPR DNA组装技术等较新的技术也继续经受类似的复杂性、易于在先前组装的构建体/载体设计上进行迭代以及速度限制(王(Wang),JW.等人,2015“CRISPR/Cas9核酸酶与吉布森组装组合用于无缝克隆(CRISPR/Cas9 nuclease combinedwith Gibson assembly for seamless cloning)”生物技术(BioTechniques),第58卷,第4期:161-170)。CRISPR克隆需要紧挨着相容的原间隔子邻近基序(PAM)靶向的功能性引导RNA的设计。靶位点内适合的PAM序列的可用性导致对基因组或构建体内可能的DNA插入定位的数量有显著的设计限制。
此外，引导RNA序列的设计和测试对多组分组装施加了显著的技术挑战。本领域技术人员将认识到，例如，并非所有gRNA序列都是功能性地，并且CRISPR DNA组装的有效实施有时可能会需要多个gRNA序列变体的设计和验证。这些限制在多组分组装中特别麻烦，其中单一gRNA序列无法成功产生期望的修饰可以触发对重新设计不再落入原始克隆计划内的后续组装组分的需要。因此，应用需要多个定制引导RNA用于多组分组装的每个接合的技术也可能会非常昂贵、麻烦且不切实际。
模块化CRISPR标签组装载体和使用其的方法
在一些实施例中，本公开教导了用于DNA组装的方法，所述方法克服了与上文所描述的上述传统技术相关联的许多限制。在一些实施例中，本公开还教导了用于与本发明的方法一起使用的模块化CRISPR组装构建体、组合物和试剂盒。
在一些实施例中，本公开教导了包括一或多个CRISPR多克隆位点(cMCS)的DNA构建体。在一些实施例中，本公开的cMCS表示所描述的DNA构建体的仅一部分(即，构建体的仅一部分根据本公开的方法可容易编辑)。在其它实施例中，本公开的cMCS位于整个构建体内的关键位置，使得整个DNA构建体可容易编辑。因此，在一些实施例中，模块化cTAG载体的所有功能性部分(例如，组装所需的所有起点、标志物、货物、元件)都包括在插入DNA部分内并且可以通过本公开的基因编辑方法容易地交换。
在一些实施例中，本公开的cMCS包括一或多个克隆标签(cTAG)，每个cTAG包括至少一个经过验证的CRISPR靶向位点。在一些实施例中，本公开的cMCS进一步包括DNA插入部分，每个DNA插入部分侧接有一对cTAG，使得用靶向一或多个cTAG的一或多种CRISPR核酸内切酶消化cMCS将释放所述被侧接的插入部分，从而允许相容的供体DNA部分的插入。
本说明书的图2和3展示了根据本公开的方法的模块化CRISPR组装质粒构建体的实施例。所公开的示例质粒含有一系列DNA插入物(图2A中的部分1-8)，每个DNA插入物侧接有图2A中的一对cTAG(标签A-H)。用适当的CRISPR/引导序列复合物消化本实例的cTAG A和B将释放质粒的部分2，从而允许具有期望特性的置换部分2插入物的插入。
本领域技术人员将立即认识到当前描述的载体系统的优点，所述载体系统允许在体内和在体外对载体进行序列特异性模块化克隆/编辑。下文的各部分将概述公开的模块化克隆载体的各个方面以及其到分子生物学、基因疗法和基因编辑的各种应用。
模块化CRISPR载体插入部分
在一些实施例中，本公开的插入部分是用于CRISPR消化后的同源重组插入的供体DNA序列。因此，在一些实施例中，本公开的插入部分序列包括感兴趣插入序列，所述感兴趣插入序列侧接有与消化的模块化CRISPR构建体的末端具有足够同源性的序列，以触发序列的同源重组、杂交和插入。
在其它实施例中，本公开的插入部分是能够通过粘性末端杂交和连接(例如，在Cpf1消化、限制性酶消化、吉布森组装或其它基于杂交的组装(包含LCR)之后)的供体DNA序列。因此，在一些实施例中，本公开的插入部分序列包括感兴趣插入序列，所述感兴趣插入序列侧接有与消化的模块化CRISPR构建体的末端具有足够同源性的序列，以允许粘性末端的杂交。
在又其它实施例中，本公开的插入部分是用于平端连接的供体DNA序列。
在一些实施例中，本公开的模块化CRISPR DNA构建体与任何插入部分序列相容。因此，本发明载体的各部分可以包括但不限于可选择标志物、复制起点、启动子、终止子序列；其它调节序列、条形码、重组位点或用户的其它感兴趣序列。在一些实施例中，本公开的插入部分可以包括用于触发同源重组和插入到一或多个遗传基因座中的同源性序列。在一些实施例中，所述同源重组插入部分将在也将通过重组事件插入基因组中的其它插入部分之前和之后。
在一些实施例中，本公开教导了每个插入部分包括单个序列(例如，仅启动子或仅感兴趣基因，参见图2A的部分8)。在其它实施例中，本公开教导了一或多个插入部分可以含有多个要素，如启动子-感兴趣基因(GOI)组合、多亚基嵌合蛋白融合物或甚至整个构建体(参见图2A的部分5，包括启动子-GOI-终止子组合)。
在一些实施例中，本公开教导了未组合的单独插入部分。即，在一些实施例中，本公开教导了一或多个未连接的插入部分(参见图2A，右侧示出了未组合的插入部分的列表)。在一些实施例中，本公开教导了将所述多个部分组装成一或多个模块化CRISPR构建体的方法。在一些方面，本公开教导了用于组装大模块化构建体的试剂盒。
在其它实施例中，本公开教导了部分或完全组装的模块化CRISPR DNA构建体。例如，在一些实施例中，本公开教导了包括1个、2个、3个、4个、5个、6个、7个、8个、9个、10个、11个、12个、13个、14个、15个、16个、17个、18个、19个、20个、21个、22个、23个、24个、25个、26个、27个、28个、29个、30个、31个、32个、33个、34个、35个、36个、37个、38个、39个、40个、41个、42个、43个、44个、45个、46个、47个、48个、49个、50个、51个、52个、53个、54个、55个、56个、57个、58个、59个、60个、61个、62个、63个、64个、65个、66个、67个、68个、69个、70个、71个、72个、73个、74个、75个、76个、77个、78个、79个、80个、81个、82个、83个、84个、85个、86个、87个、88个、89个、90个、91个、92个、93个、94个、95个、96个、97个、98个、99个、100个或更多个组装的插入部分以及其间的任何范围的模块化CRISPR DNA构建体。本公开还教导了包括所述插入部分的试剂盒。
在一些实施例中，所述组装的或部分组装的模块化CRISPR DNA构建体是线性的。在一些实施例中，所述组装的或部分组装的模块化CRISPR DNA构建体是环状的(例如，质粒)。在一些实施例中，所述组装的或部分组装的模块化CRISPR DNA构建体被整合到基因组DNA中。
在一些实施例中，本公开的构建体最初将仅含有短间隔子序列作为占位符以用于进一步克隆(参见图3C中的“填充物”序列)。在一些实施例中，插入部分占位符是小的随机化序列。在其它实施例中，本公开的载体最初将包括一或多个预先选择的插入DNA部分。例如，在一些实施例中，模块化CRISPR构建体最初将包括至少一种选择标志物和/或至少一个复制起点。
适合的可选择标志物包含但不限于赋予抗生素抗性的基因、对荧光蛋白进行编码的基因、tRNA基因、营养缺陷型标志物、毒性基因、表型标志物、反义寡核苷酸、限制性核酸内切酶、限制性核酸内切酶切割位点、酶切割位点、蛋白质结合位点以及与PCR引物序列互补的序列。
适合的抗生素抗性基因包含但不限于氯霉素抗性基因、氨苄青霉素(ampicillin)抗性基因、四环素(tetracycline)抗性基因、博莱霉素抗性基因、大观霉素(spectinomycin)抗性基因和卡那霉素抗性基因。
在本发明的某些实施例中，可逆选择标志物是毒性基因。适合的毒性基因包含但不限于ccdB基因、对结合一或多个ter位点的tus蛋白进行编码的基因、kicB基因、sacB基因、ASK1基因、ΦX174 E基因和DpnI基因。在一些实施例中，毒性可选择标志物的存在用作插入未进行或未成功的指示物。毒性可选择标志物还可以用于通过用仍在原处的毒性基因使携带未修饰的载体的细胞死亡来减少阳性细胞的未修饰的亲本载体的背景。
在本发明的方法的另外的实施例中，模块化CRISPR构建体可以包括一或多种毒性基因和一或多种抗生素抗性基因两者。
在一些实施例中，模块化CRISPR构建体最初将包括至少一个调节序列。在一些实施例中，本公开教导了包括但不限于基质连接区域、表达绝缘子序列、表达增强子序列、启动子、5'UTR、3'UTR、终止子序列、终止密码子、起始密码子等的载体。在一些实施例中，模块化CRISPR构建体最初将包括用于促进所述构建体的染色体插入的序列(例如，t-DNA边界、Cre/Lox或染色体序列的同源性末端)。在一些实施例中，定位用于染色体插入的序列以便将整个模块化CRISPR构建体插入生物体的基因组中。在其它实施例中，定位用于染色体插入的序列以便插入模块化CRISPR构建体的仅一部分(参见图3D)。
在一些实施例中，本公开的插入部分甚至可以包括另外的cTAG。通过插入部分添加cTAG可以增加可用克隆方案的复杂性并且还可以通过扩展可以被置换的可用插入部分的数量来扩展构建体的大小。
在一些实施例中，本公开的插入部分可以包括传统的克隆位点。例如，在一些实施例中，本公开教导了包括捷威重组位点、限制性位点、Cre/Lox位点或其它传统克隆位点的插入部分。在一些实施例中，本公开的插入部分可以包括用于金门(golden gate)克隆的序列。在一些实施例中，本公开的插入部分可以包括用于传统限制性酶克隆的序列。在其它实施例中，本公开的插入部分可以包括用于捷威克隆的序列。
在一些实施例中，本公开教导了从传统DNA构建体产生插入部分的方法。即，在一些实施例中，本公开教导了将cTAG添加至传统DNA构建体(例如，添加至寡核苷酸、PCR片段、质粒或其它可用DNA片段)的方法。在一些实施例中，本公开教导了将cTAG添加至单个组分如感兴趣基因(GOI)启动子的方法。在其它实施例中，本公开教导了将cTAG添加至多要素构建体的方法。
在一些实施例中，本公开教导了DNA条形码的用途。在一些实施例中，本公开的条形码是独特的一系列DNA核苷酸，当存在于DNA载体中时，所述独特的一系列DNA核苷酸可以用于在数据库中查找关于载体的信息。在一些实施例中，载体的存在可以在数据库中与载体的历史相关联，所述历史包含各种组分的来源以及载体在何时并且由谁产生。条形码还可以用于区分如当分子计数所需时在其它方面相同的DNA片或其它类似应用。
在一些实施例中，本公开的条形码可以与整个载体缔合。在其它实施例中，本公开教导了将条形码整合到插入部分中。在一些实施例中，本公开的条形码在一或多个cTAG中。在具体实施例中，插入部分中的条形码可以用于标记不同的CRISPR酶或由插入部分编码的引导RNA。在一些实施例中，对条形码进行测序可以提供将在其它方面需要对整个插入部分或整个载体/构建体进行测序的信息。
本领域技术人员将认识到用于构建插入部分的方法。例如，在一些实施例中，可以用包括所述cTAG的引物通过PCR扩增将cTAG并入到DNA分子中。在其它实施例中，可以通过传统克隆技术(例如，限制性酶、吉布森或其它组装方法)来合并cTAG。在又其它实施例中，可以通过平端连接来合并cTAG。
在一些实施例中，本公开的插入部分可以具有广泛的物种相容性谱(例如，标志物可以含有原核表达序列和真核表达序列两者以使标志物在多种生物体中有效)。在其它实施例中，本公开的插入部分被设计成对单个种/属/科/目/纲/门/界或域内的生物体具有有限的适用性。例如，在一些实施例中，复制起点部分可以能够维持仅单个种或一组种中的质粒。在其它实施例中，可以对荧光标志物进行密码子优化以跨原核域和真核域两者起作用。
在一些实施例中，Cas9核酸内切酶切割靶序列的PAM上游的3-4个核苷酸。因此，Cas9复合物对cTAG的消化可能因靶标的PAM序列或原间隔子序列的丢失而导致cTAG功能丢失。在一些实施例中，本公开教导了通过将供体插入序列设计成使得其在插入时(例如，通过插入先前丢失的PAM或原间隔子序列)重构cTAG序列。针对通过Cpf1核酸内切酶切割的序列设想了类似的配置。
图2B展示了当前公开的cTAG修复概念。用Cas9核酸内切酶切割插入部分2也导致cTAG A和B的一部分丢失。随后通过同源重组来插入插入部分2a-2d中的任何一个导致cTAG全序列的恢复。
本领域技术人员将认识到几乎无限的选项用于插入部分。前面的插入物列表旨在是说明性的并且决不应被解释为限制当前公开的方法、试剂盒和构建体的适用性。
在一些实施例中，本公开的插入部分本身可以针对CRISPR酶、催化灭活CRISPR酶或推定的CRISPR酶进行编码。
如本文所用，术语“推定的CRISPR酶”是指据信能够在体外或在宿主细胞中表现出CRISPR样功能的蛋白质。本领域技术人员将认识到可以将肽分类为推定的CRISPR酶的各种方式。在一些实施例中，将基于与一或多种已知的CRISPR酶的序列或结构同源性如此对推定的CRISPR酶进行分类。在其它实施例中，将基于推定的CRISPR酶与一或多个引导RNA相互作用的能力对其进行分类。在其它实施例中，将基于功能获得或丧失文库的的基因筛选结果对推定的CRISPR酶进行分类，在所述功能获得或丧失文库中，发现对酶进行编码的DNA会影响宿主细胞的CRISPR免疫。因此，在一些实施例中，当前公开的模块化载体可以用于筛选推定的CRISPR酶文库以鉴定一或多个宿主细胞中具有CRISPR活性的有价值的酶。在一些实施例中，本公开教导了用于通过与一或多个引导RNA序列组合地测试推定的CRISPR酶并测量靶切割程度来验证推定的CRISPR酶的高通量方法。靶切割可以通过本领域技术人员已知的任何方法来测量，包含通过测量靶基因的表达的损失或通过经由例如在凝胶上运行消化的产物、对消化的产物进行测序或运行被设计成仅扩增未消化的靶DNA的PCR反应来测量消化。
模块化CRISPR克隆标签
在一些实施例中，本公开的模块化CRISPR构建体包括一或多个克隆标签(cTAG)。在一些实施例中，本公开的模块化CRISPR构建体包括1个、2个、3个、4个、5个、6个、7个、8个、9个、10个、11个、12个、13个、14个、15个、16个、17个、18个、19个、20个、21个、22个、23个、24个、25个、26个、27个、28个、29个、30个、31个、32个、33个、34个、35个、36个、37个、38个、39个、40个、41个、42个、43个、44个、45个、46个、47个、48个、49个、50个、51个、52个、53个、54个、55个、56个、57个、58个、59个、60个、61个、62个、63个、64个、65个、66个、67个、68个、69个、70个、71个、72个、73个、74个、75个、76个、77个、78个、79个、80个、81个、82个、83个、84个、85个、86个、87个、88个、89个、90个、91个、92个、93个、94个、95个、96个、97个、98个、99个、100个或更多个cTAG。
在一些实施例中，本公开教导了每个cTAG包括至少一个经过验证的CRISPR原间隔子/PAM组合序列(“CRISPR着陆位点”)。即，在一些实施例中，cTAG包括至少一个以实验方式验证的高效CRISPR着陆位点。在一些实施例中，本公开的cTAG可以通过湿式工作台实验(例如，用靶向所述CRISPR着陆位点的CRISPR复合物在体外切割cTAG序列)来验证。在其它实施例中,cTAG验证可以从同行评审的期刊中的切割报告来假设。
在一些实施例中，本公开的cTAG包括1个、2个、3个、4个、5个、6个、7个、8个、9个、10个或更多个CRISPR着陆位点。在一些实施例中，CRISPR着陆位点彼此重叠。在其它实施例中，CRISPR着陆位点占据cTAG内的不同非重叠区域。在一些实施例中，CRISPR着陆位点可以针对Cas9或Cpf1核酸内切酶切割具有特异性。在一些实施例中，CRISPR着陆位点可以对任何其它当前的或有待发现的CRISPR核酸内切酶具有特异性。
在其它实施例中，本公开教导了可以将单个cTAG中的多个克隆位点设计成跨不同生物体起作用。因此，在一些实施例中，在缺乏或下调HR机制的生物体中，可以优选cTAGCpf1着陆位点。在其它实施例中，对于最初的体外克隆，可以优选cTAG的限制性位点，而对于选择的真核生物体中在体内发生的更复杂的编辑，可以优选Cas9或Cpf1着陆位点。
在一些实施例中，本公开教导了cTAG可以包括一或多个非CRISPR克隆序列。例如，在一些实施例中，本公开的cTAG可以包括选自由以下组成的组的一或多种要素：限制性酶位点、重组位点、拓扑异构酶位点、剪接位点和Cre-Lox位点。
在一些实施例中，适合的限制性酶位点包含但不限于由选自由以下组成的组的限制性酶识别的位点：AaII、AarI、AasI、AatII、Acc65I、AccB7I、AccI、AccIII、AciI、AclI、AcuI、AdeI、AfeI、AflII、AflIII、AgeI、AhdI、AleI、AloI、AluI、Alw21I、Alw26I、Alw44I、AlwI、AlwNI、ApaI、ApaLI、ApeKI、ApoI、AscI、AseI、AsiSI、AvaI、AvaII、AvrII、BaeI、BalI、BamHI、BanI、BanII、BbsI、BbuI、BbvCI、BbvI、BccI、BceAI、BcgI、BciVI、BclI、BcnI、BcuI、BfaI、BfiI、BfmI、BfrBI、BfuAI、BfuCI、BfuI、BglI、BglII、BlpI、Bme1390I、Bme1580I、BmgBI、BmrI、BmtI、BoxI、BpiI、BplI、BpmI、Bpu10I、Bpu1102I、BpuEI、BsaAI、BsaBI、BsaHI、BsaI、BsaJI、BsaMI、BsaWI、BsaXI、BseDI、BseGI、BseJI、BseLI、BseMI、BseMII、BseNI、BseRI、BseSI、BseXI、BseYI、BsgI、Bsh1236I、Bsh1285I、BshNI、BshTI、BsiEI、BsiHKAI、BsiWI、BslI、BsmAI、BsmBI、BsmFI、BsmI、BsoBI、Bsp19I、Bsp120I、Bsp1286I、Bsp1407I、Bsp143I、Bsp143II、Bsp68I、BspCNI、BspDI、BspEI、BspHI、BspLI、BspMI、BspPI、BspQI、BspTI、BsrBI、BsrDI、BsrFI、BsrGI、BsrI、BsrSI、BssHII、BssKI、BssSI、Bst1107I、Bst98I、BstAPI、BstBI、BstEII、BstF5I、BstNI、BstOI、BstUI、BstXI、BstYI、BstZI、BstZ17I、Bsu15I、Bsu36I、BsuRI、BtgI、BtgZI、BtsCI、BtsI、BveI、Cac8I、CaiI、CfoI、Cfr10I、Cfr13I、Cfr42I、Cfr9I、CfrI、ClaI、CpoI、Csp45I、Csp6I、CspI、CspCI、CviaII、CviKI-1、CviQI、DdeI、DpnI、DpnII、DraI、DraIII、DrdI、EaeI、EagI、Eam1104I、Eam1105I、EarI、EciI、Ecl136II、EclHKI、Eco105I、Eco130I、Eco147I、Eco24I、Eco31I、Eco32I、Eco47I、Eco47III、Eco52I、Eco57I、Eco57MI、Eco72I、Eco81I、Eco88I、Eco91I、EcolCRI、EcoNI、EcoO109I、EcoP15I、EcoRI、EcoRV、EheI、Esp3I、FatI、FauI、Fnu4HI、FokI、FseI、FspI、FspAI、GsuI、HaeII、HaeIII、HgaI、HhaI、Hin1I、Hin4I、Hin6I、HincII、HindIII、HinfI、HinP1I、HpaI、HpaII、HphI、Hpy166II、Hpy188I、Hpy188III、Hpy8I、Hpy99I、HpyAV、HpyCH4III、HpyCH4IV、HpyCH4V、HpyF10VI、Hsp92I、Hsp92II、I-PpoI、I-CreI、KasI、Kpn2I、KpnI、KspAI、LweI、MbiI、MboI、MboII、MfeI、MisI、MluI、MlyI、MmeI、MnlI、Mph1103I、MscI、MseI、MslI、MspA1I、MspI、MssI、MunI、Mva1269I、MvaI、MwoI、NaeI、NarI、NciI、NcoI、NdeI、NdeII、NgoMIV、NheI、NheI-HF、NlaIII、NlaIV、NmeAIII、NmuCI、NotI、NruI、NsbI、NsiI、NspI、OliI、PacI、PaeI、PaeR7I、PagI、PauI、PciI、PdiI、PdmI、Pfl23II、PflFI、PflMI、PfoI、PhoI、PleI、PmeI、PmlI、PpiI、PpuMI、PshAI、PsiI、Psp1406I、Psp5II、PspGI、PspOMI、PspXI、PstI、PsuI、PsyI、PvuI、PvuII、PvuII-HF、RsaI、RsrII、SacI、SacII、SalI、SalI-HF、SapI、SatI、Sau3AI、Sau96I、SbfI、ScaI、ScaI-HF、SchI、ScrFI、SdaI、SduI、SexAI、SfaNI、SfcI、SfiI、SfoI、SgfI、SgrAI、SinI、SmaI、SmiI、SmlI、SmuI、SnaBI、SpeI、SphI、SphI-HF、SspI、StuI、StyD4I、StyI、SwaI、TaaI、TaiI、TaqαI、TaqI、TasI、TatI、TauI、TfiI、TliI、Tru1I、Tru91、TseI、Tsp45I、Tsp509I、TspMI、TspRI、Tth111I、TurboNaeI、TurboNarI、Van91I、VspI、XagI、XapI、XbaI、XceI、XcmI、XhoI、XhoII、XmaI、XmaJI、XmiI、XmnI和ZraI。各方面还包含归巢核酸内切酶，如：I-SceI、I-CeuI和PI-PspI。这些酶对应的切割位点是本领域中已知的。
在一些实施例中，本公开教导了识别长度大于或等于八个核苷酸(≥8个限制酶)的位点的罕见限制性酶的用途。在一些实施例中，本公开教导了每个cTAG中的单个罕见限制性位点的用途。在其它实施例中，本公开的cTAG可以包括两个或两个以上限制性位点。下表2提供了根据本发明的cTAG的列表，每个cTAG的罕见限制性酶位点已加粗。
表2：示例cTAG序列、CRISPR着陆位点和罕见限制性酶位点(粗体序列部分是限制性位点)
Figure BDA0002979126820000401
Figure BDA0002979126820000411
在一些实施例中，用于在本发明中使用的适合的重组位点包含但不限于：attB位点、attP位点、attL位点、attR位点、lox位点、psi位点、tnpI位点、dif位点、cer位点、frt位点以及其突变体、变体和衍生物。在本发明的某些实施例中，拓扑异构酶识别位点(如果存在的话)由可以是IB型拓扑异构酶的I型拓扑异构酶识别并结合。适合的IB型拓扑异构酶类型包含但不限于真核核I型拓扑异构酶和痘病毒拓扑异构酶。在一些实施例中，适合的痘病毒拓扑异构酶类型包含但不限于由如牛痘病毒、肖普(Shope)纤维瘤病毒、ORF病毒、禽痘病毒、传染性软疣病毒和桑灯蛾昆虫痘病毒(Amsacta morreientomopoxvirus)等病毒产生或从其中分离的痘病毒拓扑异构酶。
在一些实施例中，可以根据用户偏好来安排CRISPR和非CRISPR克隆位点的cTAG布置。在一些实施例中，本公开教导了CRISPR结合位点应被安排成距插入部分最远。在一个说明性实施例中，cTAG可以从5'-3'布置如下：(部分I)-[R1-A1-C-A2-R2]-(部分II)，其中R＝限制性位点，A＝重组酶位点，并且C＝CRISPR着陆位点。在一些实施例中，C可以包含多个重叠的或顺序的CRISPR和/或限制性着陆位点。在一些实施例中，本公开的cTAG上的克隆位点的布置将是对称的(即，提供克隆位点的类型的对称顺序)。
在其它实施例中，本公开的cTAG上的克隆位点的布置可以是非对称的。例如，在另一个说明性实施例中，cTAG可以从5'-3'布置如下：(部分I)-[R1-A1-C1-C2]-(部分II)，其中R＝限制性位点，A＝重组酶位点，并且C1-2＝一或多个CRISPR着陆位点。在又其它实施例中，cTAG可以从5'-3'布置如下：i)(部分I)-[R1-C1-C2]-(部分II)、ii)(部分I)-[R1-C1]-(部分II)、iii)(部分I)-[C1-C2]-(部分II)或其反向顺序，其中R＝限制性位点，A＝重组酶位点，并且C1-2＝一或多个CRISPR着陆位点。
本领域技术人员将认识到各种cTAG布置的优点和应用。例如，在单一标签实施例中，模块化构建体将允许通过消化单个CRISPR核酸内切酶进行插入，但是由于缺少第二侧接有cTAG位点而不(没有对另外的cTAG的更多的例如进一步消化)允许去除或置换所述插入。在一些实施例中，本公开教导了插入的部分本身可以含有另外的cTAG，以扩展cMCS内的可能的插入部分定位的数量。
在其它实施例中，本公开教导了从模块化CRISPR构建体去除一或多个插入部分的方法。在一些实施例中，模块化CRISPR构建体的cTAG中两个或两个以上cTAG包括能够产生相容的末端的限制性酶结合位点。在一些实施例中，限制性酶位点是相同的。在其它实施例中，限制性酶位点是不同的，但是所述位点的所得消化产生了用于杂交和连接的相容末端。在一些实施例中，用于模块化CRISPR构建体的各部分的缺失的限制性位点被放置在两个或两个以上cTAG的其它末端上，使得所得连接的构建体仍然将维持相同的插入部分与cTAG比。
在一些实施例中，本公开教导了用于本公开的模块化CRISPR构建体内的缺失的限制性酶位点可以是导致相容末端的任何限制性酶。在其它实施例中，本公开教导了用于本公开的模块化CRISPR构建体内的缺失的限制性酶位点可以是导致相容末端的任何罕见的8个≥碱基长限制性酶。在选择的实施例中，本公开教导了用于本公开的模块化CRISPR构建体内的缺失的限制性酶位点可以是I-SceI和PI-PspI。
在一些实施例中，本公开教导了具有侧接有于每个插入部分的两个cTAG以产生cTAG对的模块化CRISPR构建体。在一些实施例中，上述cTAG对允许插入部分的选择性切割/置换。例如，如图2B所展示的，用靶向cTAG A和B的核酸内切酶消化模块化CRISPR质粒将会导致插入部分2的特异性去除。
如上文所讨论的，本公开的选择的实施例提供在核酸内切酶切割后恢复cTAG功能的置换插入部分。因此，如图2B所展示的，置换插入部分2a-2d包括将在插入模块化CRISPR质粒中后恢复cTAG A和B功能的序列。
在一些实施例中，本公开教导了cTAG还可以控制插入部分方向性。插入部分中的cTAG末端与模块化CRISPR构建体中的切割的cTAG之间的序列同源性将通过同源重组或杂交(例如，在吉布森方法中)来确定Cas9切割序列的插入方向性。Cpf1序列中的插入方向性也可以通过任一cTAG上的Cpf1粘性末端的沃森-克里克杂交来控制。
在一些实施例中，本公开还提供了替代性cTAG布置。例如，在一些实施例中，本公开的模块化CRISPR构建体可以被设计成提供用于使用嵌套cTAG的功能。
在一些实施例中，本公开教导了基于共享的重叠“标签”区域的基于组分的CRISPR组装，所述基于组分的CRISPR组装实现体内和体外多组分组装。在一些实施例中，本公开的标签包括用于促进未来从DNA构建体进行克隆或体外DNA组装的CRISPR着陆位点。如果DNA构建体整合到宿主生物体的基因组中，则预先选择的Cas9或Cpf1着陆位点可以促进容易的遗传改变。在单套实验中，组装策略使能够构建可以在含有多个数量和类型的DNA组分的多种生物体中使用的DNA质粒。
在一些实施例中，此组装策略可以用于组装和快速重新组装对包含代谢途径的任何期望的一组DNA组分进行编码的质粒。在其它实施例中，将cTAG设计到整合质粒中也可以用于将DNA组分直接换入和换出宿主生物体的基因组，从而避免需要克隆未来质粒。
cTAG序列设计算法
在一些实施例中，本公开教导了被设计成促进cTAG内的CRISPR着陆位点的算法。在一些实施例中，CRISPR着陆位点是从现有序列中鉴定的序列。因此，在一些实施例中，本公开教导了软件程序的使用被设计成基于期望的引导序列长度和用于指定的CRISPR酶的CRISPR基序序列(PAM，原间隔子邻近基序)在输入DNA序列的两条链上鉴定候选CRISPR靶序列。例如，用于具有PAM序列TTN的来自新凶手弗朗西丝菌U112的Cpf1的靶位点可以通过在输入序列和输入的反向补体上搜索5'-TTN-3'来进行鉴定。用于具有PAM序列TTTN的来自毛螺菌科细菌和氨基酸球菌的Cpf1的靶位点可以通过在输入序列和输入的反向补体上搜索5'-TTTN-3'来进行鉴定。同样，用于具有PAM序列NNAGAAW的嗜热链球菌(S.thermophilus)CRISPR1的Cas9的靶位点可以通过在输入序列和输入的反向补体上搜索5'-Nx-NNAGAAW-3'来进行鉴定。用于化脓链球菌的Cas9的PAM序列是5'-NGG-3'。
同样，用于具有PAM序列NGGNG的嗜热链球菌CRISPR的Cas9的靶位点可以通过在输入序列和输入的反向补体上搜索5'-N,—NGGNG-3'来进行鉴定。
在其它实施例中，本公开教导了从头开始设计CRISPR着陆位点的方法。如上所述，本领域技术人员将容易能够结合本公开的引导RNA来设计CRISPR着陆位点，其中所得的原间隔子序列与适合于期望的CRISPR核酸内切酶的PAM基序组合。
在一些实施例中，本公开教导了包括选自由以下组成的组的序列的cTAG：SEQ IDNO.65、66、67、68、69、70、71、72、73、74、78、79、80、81以及其组合。
由于在DNA靶位点的基因组中的多次出现可能导致非特异性基因组编辑，因此在鉴定所有潜在位点之后，本公开在一些实施例中教导了基于序列在相关参考基因组或模块化CRISPR构建体中出现的次数来滤出序列。对于通过“种子”序列(如用于Cpf1介导的切割的引导序列的前5nt)确定序列特异性的那些CRISPR酶，过滤步骤还可以滤出具有相同种子的不同序列。
在一些实施例中，算法工具还可以鉴定特定引导序列的潜在脱靶位点。例如，在一些实施例中，Cas-Offinder可以用于鉴定Cpf1的潜在脱靶位点(参见金姆等人,2016.“全基因组分析揭示了人细胞中Cpf1核酸内切酶的特异性”2016年06月在线发表)。还可以使用任何其它公开可用的CRISPR设计/鉴定工具，包含例如张实验室(Zhang lab)的crispr.mit.edu工具(参见许(Hsu)等人2013“RNA_引导的Cas9核酸酶的DNA靶向特异性(DNA targeting specificity of RNA_guided Cas9 nucleases)”自然生物技术31,827-832)。
在一些实施例中，可以允许用户选择种子序列的长度。出于穿过滤器的目的，还可以允许用户指定种子:PAM序列在基因组中的出现次数。默认的是筛选独特的序列。通过改变种子序列的长度和基因组中序列出现的次数来改变过滤水平。另外或可替代地，程序可以通过提供经过鉴定的一或多个靶序列的反向补体来提供与报告的一或多个靶序列互补的引导序列的序列。
模块化CRISPR DNA构建体克隆
在一些实施例中，本公开教导了用于使用本公开的模块化CRISPR DNA构建体制备新的重组核酸分子的方法。在一些实施例中，本公开教导了DNA部分组装的方法。下文提供了对每种方法的描述。
DNA组装方法
在一些实施例中，本公开教导了用于对DNA部分进行模块化组装的方法。在一些实施例中，本公开的DNA组装方法在体外进行。因此，在一些实施例中，本公开教导了以下步骤：i)形成包括至少两个插入部分DNA连同至少一种CRISPR复合物的混合物；以及ii)允许所述混合物在用于插入DNA的CRISPR消化的条件下温育；iii)然后杂交来自所述两个插入部分DNA中的每个插入部分DNA的消化的相容粘性末端；以及iv)将所述杂交的末端彼此连接以产生新的重组核酸。因此，在一些实施例中，本公开的插入部分DNA一起消化。在其它实施例中，本公开教导了用相同或不同的CRISPR复合物单独消化每个插入部分DNA的方法。在一些实施例中，至少一个插入部分没有被CRISPR复合物消化。在一些实施例中，本公开教导了在步骤iii)的杂交之前进行核酸外切酶处理(以用于如稍后部分中所描述的双CRISPR消化)。
在又其它实施例中，本公开教导了通过将插入部分末端暴露于ssDNA核酸外切酶并杂交所得的粘性末端、之后用聚合酶进行任选的填充并连接来进行插入部分的吉布森样连接。在一些实施例中，在ssDNA核酸外切酶处理之前将一或多个插入部分暴露于dsDNA核酸外切酶。在一些实施例中，本公开教导了已经通过一或多个CRISPR核酸内切酶消化的(例如，如稍后部分中描述的双CRISPR消化)插入部分或模块化CRISPR载体的吉布森样连接。
下文的各部分提供了展现可以组装和编辑本公开的插入部分和模块化CRISPR构建体的各种方式的一系列说明性实例。下文描述的技术列表提供了突出本公开的序列的实用性的说明性的一系列实例，但并不旨在是限制性的。本领域技术人员将认识到允许组装和编辑根据本公开的插入部分的其它技术。
在一些实施例中，本公开描述了涉及Cpf1和/或Cas9 CRISPR核酸内切酶的方法。除非在权利要求中指定，否则对这些特定CRISPR核酸内切酶的引用是说明性的，而不旨在是限制性的。本领域技术人员将立即认识到其它现有的或迄今未发现的CRISPR核酸内切酶对于本公开的构建体和方法的适用性。对Cpf1的引用可以解释为涵盖能够催化交错的DNA切割以产生粘性DNA末端的任何当前已知的或未发现的CRISPR核酸内切酶的用途。对Cas9的引用可以类似地解释为涵盖能够催化dsDNA平端切割的任何当前已知的或未发现的CRISPR核酸内切酶的用途。
体外Cpf1
在一些实施例中，本公开的体外DNA组装用如下文所述的Cpf1 CRISPR复合物进行。第一，将两个或两个以上插入部分与靶向至少两个插入部分之间共同的cTAG的Cpf1CRISPR复合物温育。在一些实施例中，插入部分在单种混合物中一起温育。在其它实施例中，插入部分在不同混合物中温育。
第二，在一些实施例中，对消化的产物进行纯化以去除活性CRISPR核酸酶。在一些实施例中，纯化涉及从消化的插入部分分离活性Cpf1复合物。在一些实施例中，这可以通过DNA纯化如凝胶或柱纯化来完成。在其它实施例中，纯化可以通过Cpf1灭活如通过热或化学灭活来完成。
第三，消化的插入部分在适合于Cpf1复合物所产生的相容粘性末端的杂交的条件下温育。然后根据任何已知的连接方法连接杂交的末端，所述任何已知的连接方法包含本公开的更早部分所描述的连接方法。
在一些实施例中，本公开教导了使用CRISPR和连接酶克隆方法(称为“CLIC”)的DNA组装(参见US16/310,895；WO/2018/013990，所述文献在此均通过全文引用的方式并入)。
在CLIC技术中，crRNA靶向性多核苷酸被设计成以反向朝向与被指定缺失的DNA插入区域(例如，多克隆位点“MCS”)的内部部分结合，以朝着去除的DNA片段外面切割。单独的crRNA靶向性多核苷酸还被设计成靶向DNA插入物(例如，感兴趣基因“GOI”)的外端，以在反应期间去除DNA结合位点。在一些实施例中，crRNA引导序列可以是相同的。
以反向朝向设计crRNA结合位点确保位点在切割过程中去除，从而允许侧接有相容序列突出端的两个DNA片段在同一反应中无缝连接。
体外Cas9
在其它实施例中，本公开的体外DNA组装用如下文所述的Cas9 CRISPR复合物进行。第一，将两个或两个以上插入部分与靶向至少两个插入部分之间共同的cTAG的Cas9CRISPR复合物温育。在一些实施例中，插入部分在单种混合物中一起温育。在其它实施例中，插入部分在不同混合物中温育。
第二，在一些实施例中，对消化的产物进行纯化以去除活性CRISPR核酸酶。在一些实施例中，纯化涉及从消化的插入部分分离活性Cas9复合物。在一些实施例中，这可以通过DNA纯化如凝胶或柱纯化来完成。在其它实施例中，纯化可以通过Cas9灭活如通过热或化学灭活来完成。
在一些实施例中，用于Cas9消化的产物的第三步骤是在适合于平端连接的条件下温育插入部分。
双CRISPR组装
在其它实施例中，本公开还教导了用于组装具有至少一个共享cTAG序列的几片CRISPR消化的插入部分(例如，组装在不同CRISPR着陆位点处消化的相容cTAG)的吉布森组装型方法。因此，在一些实施例中，本公开教导了如下文描述的双CRISPR消化组装。
第一，将两个或两个以上插入部分与靶向两个不同CRISPR着陆位点的两种CRISPR复合物温育，所述两个不同CRISPR着陆位点侧接有至少两个插入部分之间共同的前述cTAG内的每个部分。
在一些实施例中，所述两个不同CRISPR着陆位点一起消化。在其它实施例中，在单独的容器中，一个插入部分DNA用靶向一个CRISPR着陆位点的一种CRISPR复合物消化，并且另一个插入部分DNA用靶向第二CRISPR着陆靶位点的不同CRISPR复合物消化。在每种情况下，这些消化的结果将是两个插入DNA cTAG中的每个插入DNA cTAG中的共享cTAG将包括彼此至少1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19或20bp的序列重叠。
例如，在说明性实施例中，两个插入DNA部分之间的共享cTAG从5'-3'布置如下：(部分I)-[R1-C1-C2]-(部分II)，其中R＝限制性位点，C1＝第一CRISPR着陆位点，并且C2＝第二CRISPR着陆位点。在此说明性实施例中，带有3'共享cTAG的第一插入DNA将用靶向C2的CRISPR复合物消化，并且带有5'共享cTAG的第二插入DNA将用靶向C1的CRISPR复合物消化.这将导致两个DNA插入部分具有跨越C1-C2的重叠序列。
第二，在一些实施例中，对消化的产物进行纯化以去除活性CRISPR核酸酶。在一些实施例中，纯化涉及从消化的插入部分分离活性CRISPR复合物。在一些实施例中，这可以通过DNA纯化如凝胶或柱纯化来完成。在其它实施例中，纯化可以通过CRISPR灭活如通过热或化学灭活来完成。
第三，在一些实施例中，将CRISPR消化的插入部分与ssDNA核酸外切酶温育，以在所述两个插入DNA部分之间产生重叠的粘性末端。
第四，将消化的插入部分在适合于CRISPR复合物/核酸外切酶消化所产生的相容粘性末端的杂交的条件下温育。然后根据任何已知的连接方法连接杂交的末端，所述任何已知的连接方法包含本公开的更早部分所描述的连接方法。在一些实施例中，将杂交的部分与聚合酶温育以在连接之前填补任何缺失的序列缺口。
桥接组装
在其它实施例中，本公开教导了通过添加第三DNA序列对Cas9消化的部分进行吉布森组装，所述第三DNA序列包括与插入部分的消化的cTAG序列重叠的桥接序列。
在此说明性实例中，两个插入部分均用靶向同一CRISPR着陆位点的同一Cas9CRISPR复合物消化。在此实施例中，所得的消化的cTAG将不具有序列重叠。因此，在一些实施例中，第三步骤是供Cas9消化的插入部分进一步用ssDNA核酸外切酶消化以产生3'或5'突出端。然后将核酸外切酶消化的插入部分在适合于由CRISPR复合物和核酸外切酶消化的组合产生的相容粘性末端的杂交的条件下与桥接序列温育。然后根据任何已知的连接方法连接杂交的末端，所述任何已知的连接方法包含本公开的更早部分所描述的连接方法。在一些实施例中，本公开的核酸外切酶消化在第二步骤之前进行。
体外HDR
在其它实施例中，本公开教导了将Cas9或Cpf1核酸内切酶所消化的插入部分DNA的末端与HDR复合物组装由此触发所述消化的插入部分的重组的体外方法。
体内同源重组
在一些实施例中，本公开的体内DNA组装用如下文所述的Cpf1或Cas9 CRISPR复合物进行。在一个实施例中，将带有至少一个共享cTAG的两个或两个以上插入部分引入宿主细胞中。在一些实施例中，带有同源共享cTAG序列的DNA插入部分的存在将足以触发同源重组组装(例如，酵母同源重组)。
例如，在一些实施例中，可以组装所述两个插入DNA部分之间的至少一个共享cTAG序列以产生线性构建体。在此说明性实施例中，还可以将两个剩余的外部cTAG设计成与细胞内的另一个载体的cTAG重组(例如，插入现有质粒或染色体中)。在其它实施例中，可以通过所述两个插入DNA部分之间的第二共享cTAG的重组将所述两个部分进一步组装成环状构建体。组装的构建体可以用在用于组装的生物体中或者在一些实施例中可以被纯化并转化为第二生物体(例如，在酵母中组装并且随后转化为细菌)。
在其它实施例中，具有共享cTAG的一或多个插入部件可以在引入宿主细胞中之前被消化。因此，在一些实施例中，本公开教导了在被释放部分的体内组装之前将插入部分从较大载体释放的CRISPR消化。在一些实施例中，消化用Cas9进行。在其它实施例中，消化用Cpf1进行。在其它实施例中，消化用限制性核酸内切酶进行。在一些实施例中，插入部分的CRISPR消化在体外进行。在一些实施例中，在插入部分转化成组装宿主细胞之前，对消化的产物进行纯化以去除活性CRISPR核酸内切酶。
在一些实施例中，纯化步骤可以通过DNA纯化如凝胶或柱纯化来完成。在其它实施例中，纯化可以通过CRISPR灭活如通过热或化学灭活来完成。
体内连接
在一些实施例中，本公开教导了防止插入部分被CRISPR核酸内切酶重新切割的方法。在一些实施例中，本公开的插入部分可以通过DNA序列的化学修饰来防止核酸内切酶切割。例如，在一些实施例中，本公开教导了硫代磷酸寡核苷酸。
在一些实施例中，本公开的方法对多部分DNA组装体尤其有用。
本说明书的图2A提供了根据本公开的方法的多部分DNA组装的说明性实例。在本实例中，各自带有两个cTAG(标签A-H)的一系列八个DNA部分(部分1-8)在体外组合并且然后能够自组装(通过体内同源重组或通过连接，如上所述)。
DNA编辑方法
在一些实施例中，本公开教导了用于模块化CRISPR DNA构建体的编辑的方法。在一些实施例中，本公开的DNA编辑方法应用上文所述的DNA组装方法的相同原理，但这样做是出于编辑一或多个预先存在的模块化CRISPR DNA构建体的目的。
在一些实施例中，本公开的DNA编辑方法在体外进行。因此，在一些实施例中，本公开教导了以下步骤：i)形成包括模块化CRISPR DNA构建体和至少一个插入DNA部分连同至少一种CRISPR复合物的混合物；以及ii)允许所述混合物在用于插入DNA的cTAG以及其对应的模块化CRISPR DNA构建体cTAG的CRISPR消化的条件下温育、之后iii)杂交通过消化上述cTAG中的每个cTAG产生的相容粘性末端(如果是Cpf1的话)；以及iv)将所述杂交的末端(或平端，如果使用Cas9的话)彼此连接以产生新的重组核酸。在一些实施例中，核酸外切酶处理在步骤iii)的杂交之前进行(以用于如稍后部分中所描述的双CRISPR消化)。在一些实施例中，本公开的消化针对插入部分DNA和模块化CRISPR DNA构建体单独进行。在一些实施例中，仅模块化CRISPR DNA构建体用CRISPR复合物消化。
体外Cpf1
在一些实施例中，本公开的体外DNA编辑方法用如下文所述的Cpf1 CRISPR复合物进行。第一，将模块化CRISPR DNA构建体和至少一个插入DNA部分与靶向插入部分的cTAG以及其在模块化CRISPR DNA构建体内的对应标签的Cpf1 CRISPR复合物温育。在一些实施例中，模块化CRISPR DNA构建体和插入部分DNA的消化在单独的反应中进行。
第二，在一些实施例中，对消化的产物进行纯化以去除活性CRISPR核酸酶。在一些实施例中，纯化涉及从消化的核甘酸分离活性Cpf1复合物。在一些实施例中，这可以通过DNA纯化如凝胶或柱纯化来完成。在其它实施例中，纯化可以通过Cpf1灭活如通过热或化学灭活来完成。
第三，将消化的模块化CRISPR DNA构建体和插入部分在适合于Cpf1复合物所产生的相容粘性末端的杂交的条件下温育。然后根据任何已知的连接方法连接杂交的末端，所述任何已知的连接方法包含本公开的更早部分所描述的连接方法。
体外Cas9
在其它实施例中，本公开的体外DNA编辑方法用如下文所述的Cas9 CRISPR复合物进行。第一，讲模块化CRISPR DNA构建体和至少一个插入DNA部分与靶向插入部分的cTAG以及其在模块化CRISPR DNA构建体内的对应标签的Cas9 CRISPR复合物温育。在一些实施例中，模块化CRISPR DNA构建体和插入部分DNA的消化在单独的反应中进行。
第二，在一些实施例中，对消化的产物进行纯化以去除活性CRISPR核酸酶。在一些实施例中，纯化涉及从消化的核苷酸分离活性Cas9复合物。在一些实施例中，这可以通过DNA纯化如凝胶或柱纯化来完成。在其它实施例中，纯化可以通过Cas9灭活如通过热或化学灭活来完成。
在一些实施例中，用于Cas9消化的产物的第三步骤是在适合于平端连接的条件下温育插入部分。
吉布森编辑
在其它实施例中，本公开还教导了用于编辑含有完整重叠cTAG序列的CRISPR消化的构建体和/或未消化的插入部分的序列的吉布森组装型方法。因此，在一些实施例中，第三步骤是供Cas9消化的模块化CRISPR DNA构建体和一或多个插入部分被进一步用ssDNA核酸外切酶消化以产生3'或5'突出端。在一些实施例中，本公开教导了在ssDNA消化之前用于缩短非CRISPR消化的插入部分的dsDNA核酸外切酶消化。
然后将核酸外切酶消化的DNA区段在适合于由CRISPR复合物和核酸外切酶消化的组合产生的相容粘性末端的杂交的条件下温育。然后根据任何已知的连接方法连接杂交的末端，所述任何已知的连接方法包含本公开的更早部分所描述的连接方法。在一些实施例中，将杂交的DNA与聚合酶温育以在连接之前填补缺失的DNA区段。在一些实施例中，本公开的核酸外切酶消化在CRISPR灭活步骤之前进行。
在一些实施例中，消化的序列的连接可以在体外发生。
在其它实施例中，本公开教导了将Cas9或Cpf1核酸内切酶所消化的模块化CRISPRDNA构建体的末端以及至少一个未消化的插入物与HDR复合物组装由此触发所述消化的模块化CRISPR DNA构建体和至少一个插入DNA部分的重组的体外方法。
在本公开的DNA编辑方法的一些实施例中，DNA插入部分包括在第二模块化CRISPRDNA构建体内。因此，在一些实施例中，本公开的DNA编辑方法包括将DNA插入部分从一个模块化CRISPR DNA构建体转移到另一个模块化CRISPR DNA构建体。
模块化CRISPR DNA构建体用于调节宿主细胞中的CRISPR活性的用途
在一些实施例中，本公开教导了用于调节细胞中CRISPR活性的组合物和方法。因此，在一些实施例中，本公开教导了重组模块化CRISPR DNA构建体，其中所述构建体包括用于一或多种CRISPR功能调节剂的核酸。
如本文所用，术语“CRISPR功能调节剂”应当被宽泛地解释成是指当存在于额外的染色体载体中时或者当整合到宿主细胞的基因组中时导致宿主细胞中的CRISPR活性的修饰(例如，增加或减少)的任何序列。在一些实施例中，本公开教导了CRISPR功能调节剂选自由以下组成的组：复制起点、可选择标志物、抗CRISPR蛋白、启动子、终止子、dCas蛋白、dCpf1蛋白、条形码、Cas9蛋白、Cpf1蛋白、DNA供体和促进多重化的蛋白质。
在具体实施例中，CRISPR功能调节剂是抗CRISPR蛋白(帕鲁克(Pawluk)等人,“抗CRISPR：发现、机制和功能(Anti-CRISPR:discovery,mechanisms and function)”自然评论：微生物学(Nature Reviews:Microbiology)第16卷,2018年1月,第12-16页)。CRISPR-Cas适应性免疫系统广泛存在于细菌和古细菌中。然而，最近的研究示出，CRISPR系统对细菌免疫有最小的长期进化影响，从而表明存在帮助噬菌体和其它移动遗传元件逃避CRISPR-Cas免疫的抗CRISPR因子。
迄今为止，已有针对I型和II型CRISPR-Cas系统描述的21个独特的抗CRISPR蛋白家族。在过去的几年中，通过使用遗传、生化和结构研究的结合，已经确定了几种抗CRISPR蛋白的作用机制。一些抗CRISPR阴性蛋白通过干扰DNA结合来调节CRISPR酶的功能。其它抗CRISPR蛋白触发CRISPR酶的二聚化，从而减少针对基因编辑的可用性。又其它抗CRISPR蛋白阻断CRISPR酶的核酸内切酶活性，从而减少其制造双链DNA断裂的能力。在一些实施例中，本公开教导了抗CRISPR蛋白通过影响天然CRISPR系统或通过进一步修饰外源性添加的CRISPR复合物的效果来调节宿主CRISPR活性的活性的用途。
在一些实施例中，CRISPR功能调节剂用于“微调”宿主细胞中的CRISPR活性。在其它实施例中，CRISPR功能调节剂用于减少宿主细胞中的CRISPR活性。在其它实施例中，CRISPR功能调节剂用于增加宿主细胞中的CRISPR活性。
在一个说明性实例中，本公开教导了抗CRISPR酶用于提高宿主细胞的转化率的用途。在一些物种中，CRISPR表示转化障碍。本公开教导了通过使用抗CRISPR蛋白来消除基于CRISPR的先天免疫可以使一些微生物物种适于基因操纵。最近的报告描述了源自噬菌体的各种抗CRISPR蛋白(邦迪-德诺米(Bondy-Denomy)等人,2013自然493:429-432)。使用抗CRISPR蛋白来克服转化障碍有许多优点：第一，通过基因组DNA序列分析可以容易地确定宿主生物体中CRISPR系统的存在。第二，抗CRISPR的共表达不需要宿主基因组的先验操纵。第三，抗CRISPR蛋白的诱导型表达允许对质粒的阴性选择(即，当抗CRISPR蛋白表达被关闭时，质粒以CRISPR依赖性方式被破坏)。
在一个实施例中，对一或多种抗CRISPR蛋白进行编码的质粒以及抗生素抗性基因和一或多个感兴趣基因被转化为新的宿主，并且转化子是根据抗生素抗性选择的。抗抗生素克隆的存在和当抗CRISPR基因表达被关闭时抗生素抗性的丧失是指示转化障碍已被有效地消除的表型。在第二实施例中，抗CRISPR蛋白与所述质粒共转化以确保在基因表达建立之前，质粒不受活性CRISPR系统的限制。表3中提供了与本公开的组合物和方法相容的抗CRISPR蛋白的非限制性列表。
表3：本公开的抗CRISPR蛋白的非限制性列表
Figure BDA0002979126820000511
Figure BDA0002979126820000521
Figure BDA0002979126820000531
Figure BDA0002979126820000541
模块化CRISPR DNA构建体在CRISPRi和CRISPRa应用中的用途
介绍/背景
代谢工程在很大程度上依赖于分子的代谢、调节和分解代谢中直接或间接涉及的关键基因的改变。例如，减少竞争性代谢途径中使用的虹吸辅因子或代谢前体所涉及的基因表达通常是有用的。但是，代谢组的复杂性使得难以先验地预测下调所需的正确基因和下调需要的最佳数量，以在限制细胞毒性的同时提高产率。代谢物产量上的上位或非相加、天然或多个基因变化进一步使对可能会有益的多组变化的预测变得复杂。
因此，强烈需要使能够对调节网络进行快速高通量取样的技术和方法。有许多策略使能够改变表达。改变基因的拷贝数量、变化基因的优选密码子使用、变化驱动基因的启动子的强度、改变mRNA的核糖体结合位点以及调节mRNA转录物都已经被用于影响基因表达的变化。然而，这些变化中的每个变化都依赖于做出基因改变。根据所使用的生物体，基因组编辑是耗时、费力且常常效率低下的过程。这既限制了可以测试的变化的范围，又在许多生物体中阻止了多种基因组变化的同时多重化。
作为基因改变的替代方案，许多研究人员已经使用催化灭活Cas9(dCas9)来阻遏原核生物体中的转录。在如谷氨酸棒状杆菌等原核生物中，针对基因的启动子或开放阅读框(ORF)的dCas9可以阻断转录起始或伸长。最近的出版物也已经提出了通过将转录激活亚基与催化灭活dCas9基因融合来上调基因的能力。靶向用于下调的不同基因的引导RNA文库已经用于筛选或选择有益的变化。最后，原核基因的激活在大肠杆菌中以低通量得到展现。
但是，需要将这些技术整合到成功的高通量代谢工程活动中。进一步地，需要将这些不同的策略组合并应用到如谷氨酸棒状杆菌等非典型生物体中。向代谢工程活动应用大的dCas9指导型引导RNA文库组的需求尚未得到满足。
本公开描述了使用Cas9或dCas9作为筛选工具，所述筛选工具将极大地增加可以测试的转录扰动的数量，同时减少需要做出的费力的基因改变的数量。本公开通过利用当前公开的模块化CRISPR DNA构建体用于CRISPRi/CRISPRa应用的能力和灵活性来解决这些问题。这些方法可以单独使用或者在某些实施例中可以与用于基因修饰的CRISPR方法组合以调节来自宿主基因组序列的表达并且也修饰基因组序列。例如，本公开的构建体可以包含对DNA进行编码的活性和非活性CRISPR蛋白两者，或者可以产生多个构建体，其中不同的构建体含有不同的CRISPR(例如，非活性和活性)CRISPR形式。
技术综述
在一些实施例中，本公开教导了通过CRISPRi(CRISPR干扰)和CRISPRa(CRISPR激活)技术来调节宿主细胞基因的表达的方法。在一些实施例中，当前公开的技术利用已经发生突变以不再生成双DNA链断裂的催化灭活(即，核酸酶去激活)CRISPR核酸内切酶，但是所述催化灭活的CRISPR核酸内切酶仍然能够通过其对应的引导RNA与DNA靶位点结合。在一些实施例中，本公开将这些催化灭活CRISPR酶称为“死CRISPR”或“dCRISPR”酶。修饰语“死”也可以用于指特定的CRISPR酶，如死Cas9(dCas9)或死Cpf1(dCpf1)。
不希望受任何理论的束缚，发明人相信此技术的dCRISPR酶通过经由引导RNA将催化灭活dCRISPR酶募集到靶DNA序列而起作用，由此允许dCRISPR酶针对特定基因与宿主细胞的转录机制相互作用。
在一些实施例中，本公开的CRISPRi方法利用dCRISPR酶来占据转录所必需的靶DNA序列，从而阻断靶向的基因的转录(L.S.齐(Qi)等人,“再利用CRISPR作为用于基因表达的序列特异性控制的RNA引导平台.”细胞.152,1173-1183(2013)；还参见L.A.吉尔伯特(Gilbert)等人,“CRISPR介导的模块化RNA引导的真核生物转录调节(CRISPR-MediatedModular RNA-Guided Regulation of Transcription in Eukaryotes).”细胞.154,442-451(2013))。在其它实施例中，本公开的CRISPRi方法利用与一或多个转录阻遏结构域翻译融合或以其它方式拴系的dCRISPR酶或替代性地利用能够将转录阻遏结构域募集到靶位点(例如，通过适配子拴系，如下文所讨论的)的经过修饰的引导RNA。
在一些实施例中，本公开的CRISPRa方法采用与不同转录激活结构域翻译融合或以其它方式拴系的dCRISPR酶，所述不同转录激活结构域可以通过引导RNA指向启动子区域。(参见A.W.程(Cheng)等人,“RNA引导的转录激活因子系统CRISPR-on对内源基因的多重化激活(Multiplexed activation of endogenous genes by CRISPR-on,an RNA-guidedtranscriptional activator system).”细胞研究(Cell Res.)23,1163-1171(2013)；还参见L.A.吉尔伯特等人,“基因组规模的CRISPR介导的基因阻遏和激活控制.”细胞.159,647-661(2014))。在其它实施例中，本公开的CRISPRa方法利用经过修饰的引导RNA，所述经过修饰的引导RNA募集另外的转录激活结构域以上调靶基因的表达(例如，通过适配子拴系，如下文所讨论的)。
在又其它实施例中，当前公开的发明还设想利用dCRISPR酶和引导RNA来募集其它调节因子以靶向DNA位点。除了募集转录阻遏或激活结构域之外，如上文所讨论的，可以修饰本公开的dCRISPR酶和引导RNA，以募集具有范围为DNA甲基化、染色质重塑、泛素化、sumo化(sumoylation)的活性的蛋白质。因此，在一些实施例中，可以修饰本公开的dCRISPR酶和引导RNA以募集具有以下的因子：甲基转移酶活性、脱甲基酶活性、脱氨基作用活性、歧化酶活性、烷基化活性、脱嘌呤活性、氧化活性、嘧啶二聚体形成活性、整合酶活性、转座酶活性、重组酶活性、聚合酶活性、连接酶活性、解旋酶活性、光解酶活性、糖基化酶活性、乙酰转移酶活性、脱乙酰酶活性、激酶活性、磷酸酶活性、泛素连接酶活性、去泛素化活性、腺苷酰化活性、脱腺苷化活性、sumo化活性、去sumo化活性、核糖基化活性、去核糖基化活性、豆蔻酰化活性、重塑活性、蛋白酶活性、氧化还原酶活性、转移酶活性、水解酶活性、裂解酶活性、异构酶活性、合酶活性、合成酶活性、去豆蔻酰化活性、胞苷脱氨酶活性以及其任何组合
在其它实施例中，可以修饰本公开的dCRISPR酶和引导RNA以募集一或多个标志物基因/组合物，如荧光蛋白、金颗粒、放射性同位素、GUS酶或能够被检测到的其它已知的生物或合成组合物。此最后实施例将允许研究人员对宿主细胞的基因组的区域进行加标签和跟踪。如本文所用，术语“顺式调节因子”是指可以被本公开的dCRISPR或引导RNA募集的任何生物或合成组合物。
高通量CRISPRi和CRISPRa载体
在一些实施例中，本公开教导了用于宿主细胞的高通量CRISPRi/CRISPRa基因工程的载体、试剂盒和方法。在一些实施例中，本公开利用此应用中讨论的模块化CRISPR构建体的力量来进行高效的全基因组基因表达修饰(增加或减少)。在一些实施例中，本公开的模块化CRISPR构建体能够一次调节(增加或减少)1个、2个、3个、4个、5个、6个、7个、8个、9个、10个、11个、12个、13个、14个、15个、16个、17个、18个、19个、20个、21个、22个、23个、24个、25个、26个、27个、28个、29个、30个、31个、32个、33个、34个、35个、36个、37个、38个、39个、40个、41个、42个、43个、44个、45个、46个、47个、48个、49个、50个、51个、52个、53个、54个、55个、56个、57个、58个、59个、60个、61个、62个、63个、64个、65个、66个、67个、68个、69个、70个、71个、72个、73个、74个、75个、76个、77个、78个、79个、80个、81个、82个、83个、84个、85个、86个、87个、88个、89个、90个、91个、92个、93个、94个、95个、96个、97个、98个、99个、100个或更多个基因的表达。
在一些实施例中，本公开教导了包括针对本公开的CRISPRi或CRISPRa系统进行编码的核酸的模块化CRISPR构建体。在一些实施例中，本公开的模块化CRISPR构建体包括i)针对dCRISPR酶进行编码的第一核酸序列和ii)针对能够将dCRISPR酶募集到DNA靶位点的引导RNA进行编码的第二核酸。在其它实施例中，本公开教导了CRISPRa/CRISPRi系统的一或多个部分可以从载体中排除(如果所述一或多个部分已经存在于宿主中的话)或者以其它方式由单独的载体提供。因此，在一些实施例中，模块化CRISPR构建体将不针对dCRISPR酶进行编码。在其它实施例中，所述模块化CRISPR构建体将不针对引导RNA进行编码。
本领域技术人员将理解，本公开的模块化CRISPR构建体可以针对多于一种dCRISPR酶和/或多于一个引导RNA进行编码(参见例如图18A-B)。
在一些实施例中，本公开的模块化CRISPR构建体的一或多个插入部分中含有针对dCRISPR酶和/或引导RNA进行编码的核酸。因此，在一些实施例中，本公开的模块化CRISPR构建体允许用户快速且高效地修饰构建体以加上或减去针对不同引导RNA(例如，靶向不同基因或对能够募集如上文所讨论的不同顺式调节因子的适配子进行编码的引导RNA)进行编码或对不同的dCRISPR酶(例如，dCas9或dCpf1或与如上文所讨论的各种顺式调节因子的dCRISPR蛋白融合物)进行编码的插入部分。
在一些实施例中，本公开教导了模块化CRISPR构建体的插入部分包括仅单个编码序列。即，在一些实施例中，本公开的插入部分将仅针对单个dCRISPR进行编码或将仅针对单个引导RNA进行编码。在其它实施例中，本公开教导了具有期望的基因组合的插入部分。例如，在一些实施例中，可以设计插入部分以针对多个引导RNA如在生物合成途径中靶向两个或两个以上基因的引导RNA进行编码。在其它实施例中，单个插入部分可以被设计成针对多于一种dCRISPR酶进行编码。在一些实施例中，其它基因也可以与引导RNA或dCRISPR酶组合地被编码。
在一些实施例中，本公开的插入部分被设计成针对可以与由构建体编码的一或多个组分相关联的(即，用于展现由构建体编码的一或多个组分的存在的)可选择标志物进行编码，例如，以鉴定含有插入部分的细胞。因此，在一些实施例中，插入部分可以被设计成包括可选择标志物连同针对特定dCRISPR酶进行编码的核酸序列。本公开进一步设想了可选择标志物用于选择插入部分的块，例如靶向特定生物合成途径的引导RNA的块或针对一或多个顺式调节因子进行编码的插入部分的块的用途。
在一些实施例中，本公开的插入部分被设计成是独立的，包括表达插入部分所必需的所有元件。即，在一些实施例中，本公开的插入部分将含有供插入部分被宿主细胞机制表达所必需的所有必需启动子和/或终止子(不必计算维持质粒所需的复制起点或可选择标志物)。在其它实施例中，本发明的插入部分的表达将依赖于位于不同插入部分中的启动子或终止子序列，如紧挨着基因编码插入部分上游放置的启动子或终止子序列或放置在单个插入部分中或跨越多于一个插入部分的针对多顺反子mRNA进行编码的核酸开头的启动子或终止子序列。
在一些实施例中，本公开教导了诱导型启动子用于驱动模块化CRISPR构建体的插入部分中的一或多个插入部分的表达的用途。因此，在一些实施例中，本公开教导了诱导型启动子用于驱动针对dCRISPR酶进行编码的核酸的表达的用途。在其它实施例中，本公开教导了诱导型启动子用于驱动针对引导RNA进行编码的核酸的表达的用途。
在一些实施例中，模块化cTAG载体的所有功能性部分(例如，组装所需的所有起点、标志物、货物、启动子、终止子和所有元件)都包括在插入DNA部分内并且可以通过本公开的DNA组装或基因编辑方法容易地交换。因此，在具体实施例中，当前公开的模块化构建体允许快速切换和测试与对dCRISPR酶进行编码的核酸可操作地连接的不同启动子或终止子。在其它具体实施例中，当前公开的模块化构建体允许快速切换和测试与对sgRNA或crRNA/tracrRNA或引导RNA或CRISPR酶进行编码的核酸可操作地连接的不同启动子或终止子。
复制起点
在一些实施例中，本公开教导了复制起点用于维持(即，继续复制)一或多个物种中的质粒的用途。本领域技术人员将熟悉各种可用的复制起点序列。细菌、古细菌、真核和多细胞生物体的复制起点的常见特征在以下中讨论：莱纳德(Leonard)和梅查利(Mechali),“DNA复制起点(DNA replication Origins)”冷泉港生物学展望(Cold SpringHarb Perspect Biol)2013年10月；5(10)。表4中提供了常见复制起点的非限制性列表。
图4—常见载体和其复制起点的列表。
常见载体 复制数量+ ORI 对照
pUC 约500-700 pMB1(衍生) 宽松
pBR322 约15-20 pMB1 宽松
pET 约15-20 pBR322 宽松
pGEX 约15-20 pBR322 宽松
pColE1 约15-20 ColE1 宽松
pR6K 约15-20 R6K* 严格
pACYC 约10 p15A 宽松
pSC101 约5 pSC101 严格
pBluescript 约300-500 ColE1(衍生)和F1** 宽松
pGEM 约300-500 pUC和F1** 宽松
拴系顺式调节因子(转录调节剂)
在一些实施例中，本公开教导了转录调节剂的用途。因此，在一些实施例中，顺式调节因子是转录调节剂。在一些实施例中，基于转录调节因子进一步阻遏或替代性地激活本公开的CRISPRi/CRISPRa方法所靶向的基因的表达的能力来选择转录调节剂。在一些实施例中，本公开教导了将转录调节剂与dCRISPR酶拴系或翻译融合(即，通过使用融合构建体)。
融合构建体通常可以使用标准技术来制备。例如，对肽组分进行编码的DNA序列可以单独组装并且连接到如插入部分等适合的构建体中。连接的DNA序列与适合的转录或翻译调节元件可操作地连接。对一种肽组分进行编码的DNA序列的5'或3'端在有或没有肽接头的情况下与对第二肽组分进行编码的DNA序列的3'或5'端分别连接，使得序列的阅读框同相。这允许翻译成保留两种组分肽的生物活性的单一融合蛋白。
在一些实施例中，dCRISPR酶和转录调节剂结构域通过肽接头连接。可以采用肽接头序列来将第一肽组分和第二肽组分分开足够的距离，以确保每个肽折叠成其二级结构和三级结构。这种肽接头序列使用本领域熟知的标准技术并入到融合蛋白中。适合的肽接头序列可以基于以下因素进行选择：(1)其能够采用灵活的延伸构象；(2)其不能采用可以与第一肽和第二肽上的功能性区域相互作用的二级结构；以及(3)缺乏可以与肽功能性区域反应的疏水残基或带电残基。在某些实施例中，肽接头序列含有Gly、Asn和Ser残基。还可以在接头序列中使用其它近中性氨基酸，如Thr和Ala。
在一些实施例中，本公开教导了蛋白质-蛋白质相互作用结构域用于将转录调节剂结构域与dCRISPR拴系的用途。因此，在一些实施例中，dCRISPR酶的序列与第一蛋白质-蛋白质相互作用结构域(PP1)翻译融合，所述PP1能够与和转录调节剂(或其它顺式调节因子)翻译融合的第二蛋白质-蛋白质相互作用结构域(PP2)二聚化。当被表达时，dCRISPR-PP1和PP2-转录调节剂中的每一个都将二聚化，从而将转录调节剂募集到DNA靶位点。本领域技术人员将意识到使用天然存在的或合成的蛋白质-蛋白质相互作用结构域来产生体内二聚体的方法。(参见吉塞克(Giescke)等人,2006“通过改组Cys2His2锌指创建的合成蛋白质-蛋白质相互作用结构域(Synthetic protein-protein interaction domains createdby shuffling Cys2His2 zinc-fingers).”分子系统生物学(Mol Syst Biol)2:2006.0011)。
在其它实施例中，本公开还教导了聚源能够募集一或多个顺式调节因子的RNA适配子的经过修饰的引导RNA。本公开的RNA适配子可以与引导RNA的5'或3'端可操作地连接并且被设计成不影响dCRISPR与DNA靶位点的结合。相反，RNA适配子提供了另外的系链以从其中募集一或多个顺式调节因子如转录调节剂。
在一些实施例中，本公开教导了被设计成直接与一或多个顺式调节因子相互作用的定制化RNA适配子。在其它实施例中，本公开教导了靶向特定序列的已知适配子的用途。因此，在一些实施例中，本公开设想了聚源经过验证的RNA适配子的引导RNA，所述经过验证的RNA适配子然后与其天然靶标结合，所述天然靶标进而与一或多个顺式调节因子(即，引导_RNA-适配子-适配子_靶标-顺式_调节_因子)翻译融合。在一些实施例中，合并RNA适配子以拴系顺式调节因子的引导RNA被称为支架RNA(scRNA)。(扎拉坦(Zalatan)JG等人“用CRISPR RNA支架工程化复杂的合成转录程序.”细胞.2015；160:339-350)。scRNA是通过用正交地起作用的蛋白质结合RNA适配子延伸引导RNA序列来设计的。每个scRNA可以对信息进行编码以用于DNA靶标识别和募集特定阻遏蛋白或激活因子蛋白。通过以模块化方式改变DNA靶向序列或RNA适配子，多个dCas9-scRNA可以同时激活或阻遏同一细胞中的多个基因
例如，被称为协同激活介体(SAM)系统的改进是通过将MS2适配子添加到引导RNA来实现的。MS2适配子被设计成募集与p65AD和热休克因子1(HSF1)融合的同源MS2外壳蛋白(MCP)(多明格斯(Dominguez)等人,2016“超越编辑：再利用CRISPR-Cas9进行精确基因组调节和探寻(Beyond editing；repurposing CRISPR-Cas9 for precision genomeregulation and interrogation)”自然综述：分子细胞生物学(Nat Rev Mol Cel Biol)1月17(1)5-15)。SAM技术连同dCas9-VP64一起与单独dCas9-VP64相比进一步增强了内源基因激活并且被证明同时激活了10个基因。(科纳曼(Konermann)S等人“通过工程化CRISPR-Cas9复合物进行的基因组规模转录激活(Genome-scale transcriptional activation byan engineered CRISPR-Cas9 complex).”自然.2014；517:583-588)。类似的结果可以通过使用其它经过验证的适配子-支架蛋白组合如PP7或com来实现。(扎拉坦JG等人“用CRISPRRNA支架来工程化复杂的合成转录程序.”细胞.2015；160:339-350)。
在一些实施例中，本公开还设想了能够将dCRISPR酶与一或多个顺式调节因子拴系的双面适配子的用途。本公开的双面适配子与上文所讨论的适配子类似地起作用，但是能够结合dCRISPR蛋白和顺式调节因子两者。在一个说明性实例中，dCRISPR酶将与MS2外壳蛋白结构域翻译融合，并且顺式调节元件(VP16结构域)将与PP7结构域翻译融合。双面RNA适配子将在一个末端上包括MS2结合域并且在另一个末端上暴力PP7结合域。因此，在一些实施例中，本公开的双面适配子将预期形成以下一般结构：dCRISPR-适配子_靶标-适配子_面1-适配子_面2-适配子_靶标-顺式_调节_因子。
与当前公开的发明相容的转录激活结构域的非限制性列表包含：转录调节结构域的片段和具有以下的转录调节功能的结构域的片段：VP16、VP64、VP160、EBNA2、E1A、Gal4、Oaf1、Leu3、Rtg3、Pho4、Gln3、Gcn4、Gli3、Pip2、Pdr1、Pdr3、Lac9、Tea1、p53、NFAT、Sp1(例如，Sp1a)、AP-2(例如，Ap-2a)、Sox2、NF-κB、MLL/ALL、E2A、CREB、ATF、FOS/JUN、HSF1、KLF2、NF-1L6、ESX、Oct1、Oct2、SMAD、CTF、HOX、Sox2、Sox4、VPR、RpoZ或Nanog。在一些实施例中，转录激活因子是VPR(参见基亚尼(Kiani)S.等人,“用于基因组编辑、激活和阻遏的Cas9 gRNA工程(Cas9 gRNA engineering for genome editing,activation and repression)”自然方法(Nature Methods)12,1051-1054(2015))。
与当前公开的发明相容的转录阻遏物的非限制性列表包含：Mxi1、Tbx3、KRAB(克鲁贝尔相关盒，马戈林(Margolin),J.F等人“克鲁贝尔相关盒是强效转录阻遏结构域(Kruppel-associated boxes are potent transcriptional repression domains).”美国国家科学院院刊91,4509-4513(1994))、EnR或SID、SID4X(由短肽接头连接的四个SID结构域的串联重复)、PIE-1、IAA28-RD等。
在一些实施例中，本公开的转录激活结构域包括表5的激活结构域。9-氨基酸反式激活结构域(9aa TAD)定义了大的真核转录因子超家族共同的新结构域，所述真核转录因子由酵母中的Gal4、Oaf1、Leu3、Rtg3、Pho4、GIn3、Gcn4表示并且由哺乳动物中的p53、NFAT、NF-κB和VP16表示。针对9aa TAD(针对酸性和亲水性反式激活结构域)的预测可从ExPASyTM和EMBnetTM数据库在线获得。
表5—转录激活结构域的非限制性实例。
转录因子来源 9aa TAD
P53 TAD1 ETFSDLWKL(SEQ ID NO:114)
P53TAD2 DDIEQWFTE(SEQ ID NO:115)
MLL DIMDFVLK(SEQ ID NO:116)
EA2 DLLDFSMMF(SEQ ID NO:117)
Rtg3 ETLDFSLVT(SEQ ID NO:118)
CREB RKILNDLSS(SEQ ID NO:119)
CREBaB6 EAILAELKK(SEQ ID NO:120)
Gli3 DDVVQYLNS(SEQ ID NO:121)
Gal4 DDVYNYLFD(SEQ ID NO:122)
Oaf1 DLFDYDFLV(SEQ ID NO:123)
Pip2 DFFDYDLLF(SEQ ID NO:124)
Pdr1 EDLYSILWS(SEQ ID NO:125)
Pdr3 TDLYHTLWN(SEQ ID NO:126)
引导RNA多重化系统
在一些实施例中，本公开教导了引导RNA多重化系统的用途。即，在一些实施例中，本公开教导了例如使用多个启动子或多顺反子转录物表达多于一个引导RNA的方法。在一些实施例中，本公开教导了Csy4多重系统的用途。当被过表达时，Csy4高效地切割夹在28碱基Csy4识别位点之间的gRNA。CpfI也可以处理多个gRNA。(参见村田(Murata)等人2018.,“使用CRISPR/CAS9 gRNA阵列的高度多重化基因组工程(Highly multiplexed genomeengineering using CRISPR/CAS9 gRNA arrays)”公共科学图书馆·综合(PLOS ONE)13(9):e0198714，所述文献出于所有目的以其全文并入在此)。如果Csy4或其它多重化系统没有被表达，则gRNA不能被释放，从而增加了对系统的时间和/或空间控制
在一些实施例中，本公开的引导RNA侧接有核糖核酸酶识别位点。核糖核酸酶(缩写为RNA酶)是催化RNA水解的核酸酶。核糖核酸酶可以是核糖核酸内切酶或核糖核酸外切酶。核糖核酸内切酶切割单链或双链RNA。核糖核酸外切酶通过从RNA的5'端或3'端去除末端核苷酸来降解RNA。在一些实施例中，本公开的引导RNA侧接有Csy核糖核酸酶识别位点(例如，Csy4核糖核酸酶识别位点)。Csy4是识别特定RNA序列、切割RNA并维持与上游片段结合的核糖核酸内切酶。在一些实施例中，使用Csy核糖核酸酶(例如，Csy4核糖核酸酶)来从工程化核酸转录物中释放引导RNA。因此，在一些实施例中，细胞与工程化构建体共转染，所述工程化构建体包括对侧接有Csy4或其它Cas6核糖核酸酶识别位点的引导RNA进行编码的核苷酸序列以及对Csy4或其它Cas6核糖核酸酶进行编码的工程化核酸。可替代地或另外，细胞可以稳定地表达或被修饰以稳定地表达Csy4或其它Cas6核糖核酸酶。在一些实施例中，Csy核糖核酸酶(例如，Csy4核糖核酸酶)来自绿脓假单胞菌、表皮葡萄球菌(Staphylococcus epidermidis)、强烈火球菌(Pyrococcus furiosus)或硫磺矿硫化叶菌(Sulfolobus solfataricus)。本文中设想了其它核糖核酸酶和核糖核酸酶识别位点(参见例如莫吉卡(Mojica),F.J.M.等人,CRISPR-Cas系统：细菌和古细菌中RNA介导的适应性免疫(CRISPR-Cas Systems,RNA-mediated Adaptive Immunity in Bacteria andArchaea),罗尔多夫-巴兰古(Barrangou,Rodolphe),约翰-范德欧斯特(van der Oost,John)(编),2013,ISBN 978-3-642-34657-6，其中与核糖核酸酶/识别位点有关的主题通过引用的方式并入本文)。
在一些实施例中，核糖核酸酶识别位点(例如，Csy4核糖核酸酶识别位点)的长度为10到50个核苷酸。例如，Csy核糖核酸酶识别位点的长度可以为10到40个、10到30个、10到20个、20到50个、20到40个或20到30个核苷酸。在一些实施例中，Csy核糖核酸酶识别位点的长度为10个、11个、12个、13个、14个、15个、16个、17个、18个、19个、20个、21个、22个、23个、24个、25个、26个、27个、28个、29个、30个、31个、32个、33个、34个、35个、36个、37个、38个、39个、40个、41个、42个、43个、44个、45个、46个、47个、48个、49个或50个核苷酸。在一些实施例中，Csy核糖核酸酶识别位点(例如，Csy4核糖核酸酶识别位点)的长度为28个核苷酸。本文中还设想了Csy同源物(参见例如莫吉卡,F.J.M.等人,CRISPR-Cas系统：细菌和古细菌中RNA介导的适应性免疫,罗尔多夫-巴兰古,约翰-范德欧斯特(编),2013,ISBN978-3-642-34657-6，其中与核糖核酸酶/识别位点有关的主题通过引用的方式并入本文)。也可以参考美国专利序列号US 9,745,610和美国公开申请US 2017/022499，所述文献中的每个文献出于所有目的以其全文并入在此。
表达、纯化和递送
在一些实施例中，本公开教导了载体、构建体和对CRISPR复合物进行编码的核酸序列的方法和组合物。在一些实施例中，本公开教导了用于Cas9或Cpf1蛋白的转基因或瞬时表达的质粒。在一些实施例中，本公开教导了对嵌合Cas9或Cpf1蛋白进行编码的质粒，所述嵌合Cas9或Cpf1蛋白包括用于本文中描述的其它多肽中的一或多个多肽的蛋白融合的框内序列，包含但不限于连接酶、接头和NLS。
在一些实施例中，本公开的质粒和载体将针对一或多个Cas9/Cpf1蛋白进行编码并且也对crRNA/tracrRNA/sgRNA和/或本公开的供体插入序列进行编码。在其它实施例中，可以在一或多种不同的质粒中编码工程化复合物的不同组分。
在一些实施例中，本公开的质粒可以跨多个物种使用。因此，在某些实施例中，单个质粒可以被设计成允许将插入部分引入多个物种例如多个细菌种例如谷氨酸棒状杆菌和大肠杆菌中。在其它实施例中，本公开的质粒是针对被转化的生物体定制的。在一些实施例中，本公开的序列将被密码子优化以在其基因被编辑的生物体中表达。本领域技术人员将认识到使用提供充分表达以用于基因编辑的启动子的重要性。在一些实施例中，不同物种的质粒将需要不同的启动子。
在一些实施例中，本公开的质粒和载体在感兴趣细胞中选择性地表达。因此，在一些实施例中，本申请教导了异位启动子、组织特异性启动子、发育调节启动子或诱导型启动子的用途。在一些实施例中，本公开还教导了终止子序列的用途。
在一些实施例中，本公开还教导了表达和纯化Cpf1和/或Cas9核酸内切酶蛋白的方法。在一些实施例中，本公开教导了本公开的蛋白质可以由商业上可获得的蛋白质生产和纯化试剂盒或服务中的任何一个来生产。例如，在一些实施例中，本公开教导了将Cas9和/或Cpf1克隆到带有多组氨酸(His)、谷胱甘肽s-转移酶(GST)或其它纯化标签嵌合融合物的载体中的方法。在一些实施例中，本公开教导了各种原核和真核生物体以及无细胞蛋白生产系统。例如，在一些实施例中，本公开教导了大肠杆菌BL21中的蛋白表达质粒的表达。在一些实施例中，蛋白质生产系统将是诱导型的以减少蛋白毒性的影响。例如，在一些实施例中，本公开教导了使用IPTG或阿拉伯糖诱导系统的方法。
在一些实施例中，本公开还教导了各种蛋白质纯化方案，包含亲和标签(His-镍、GST-谷胱甘肽等)。在一些实施例中，本公开教导了用于蛋白质纯化的天然和变性条件两者。
在其它实施例中，本公开教导了通过一或多个蛋白质生产服务来生产Cas9和/或Cpf1，所述蛋白质生产服务包含但不限于
Figure BDA0002979126820000641
和
Figure BDA0002979126820000642
转化
在一些实施例中，本公开教导了本文所公开的质粒和载体的转化的用途。本领域技术人员将认识到，本公开的质粒可以通过本说明书的其它部分所描述的任何已知系统转化到细胞中。例如，在一些实施例中，本公开教导了通过粒子轰击、化学转化、农杆菌属转化、纳米尖峰转化(nano-spike transformation)、电穿孔和病毒转化的转化。
在一些实施例中，可以使用包含转化、转染、转导、病毒感染、基因枪或Ti介导的基因转移的各种技术中的任何一种技术将本公开的载体引入到宿主细胞中。特定方法包含磷酸钙转染、DEAE-葡聚糖介导的转染、脂质转染或电穿孔(戴维斯(Davis),L.,狄伯纳(Dibner),M.,巴蒂(Battey),I.,1986“分子生物学基本方法(Basic Methods inMolecular Biology)”)。其它转化方法包含例如醋酸锂转化和电穿孔(参见例如吉茨(Gietz)等人,核酸研究27:69-74(1992)；伊藤(Ito)等人,细菌学杂志(J.Bacterol.)153:163-168(1983)；以及贝克尔(Becker)和瓜伦特(Guarente),酶学方法(Methods inEnzymology)194:182-187(1991))。在一些实施例中，转化的宿主细胞被称为重组宿主菌株。
在一些实施例中，本公开教导了使用本公开的96孔板机器人平台和液体处理机器对细胞的高通量转化。
在一些实施例中，本公开教导了用于使外源蛋白(Cpf1/Cas9和DNA连接酶)、RNA(crRNA/tracRNA/引导RNA)和DNA(插入DNA部分或模块化CRISPR构建体)进入细胞中的方法。先前已经描述了用于实现此目的的各种方法，包含直接转染蛋白质/RNA/DNA或DNA转化、之后在细胞内表达RNA和蛋白质(迪卡洛(Dicarlo),J.E.等人“使用CRISPR-Cas系统的酿酒酵母基因组工程(Genome engineering in Saccharomyces cerevisiae usingCRISPR-Cas systems)”.核酸研究(2013).doi:10.1093/nar/gkt135；任(Ren),Z.J.,鲍曼(Baumann),R.G.和布莱克(Black),L.W.“通过过表达的T4 DNA连接酶在体内对线性DNA的克隆：T4噬菌体hoc基因展示载体的构建(Cloning of linear DNAs in vivo byoverexpressed T4 DNA ligase:construction of a T4 phage hoc gene displayvector)”.基因195,303-311(1997)；林(Lin),S.,斯塔尔(Staahl),B.T.,阿拉(Alla),R.K.和杜德纳(Doudna),J.A.“通过对CRISPR/Cas9递送的受控定时增强的同源性指导的人类基因组工程(Enhanced homology-directed human genome engineering by controlledtiming of CRISPR/Cas9 delivery).”E生命(Elife)3,e04766(2014))。
在一些实施例中，本公开教导了用一或多种如上文所描述的选择标志物来筛选转化的细胞。在一个此类实施例中，将用包括卡那霉素抗性标志物(KanR)的载体转化的细胞平板接种于含有有效量的卡那霉素抗生素的培养基上。推测在掺有卡那霉素的培养基上可见的菌落形成单位已将载体盒并入其基因组中。可以通过PCR、限制性酶分析和/或对相关插入位点的测序来确认期望的序列的插入。
在其它实施例中，本公开的一部分或全部复合物可以直接递送到细胞。因此，在一些实施例中，本公开教导了本公开的多肽和核酸的表达和纯化。本领域技术人员将认识到用于纯化蛋白质和核酸的多种方式。在一些实施例中，多肽可以通过诱导型或组成型蛋白生产系统如细菌系统、酵母系统、植物细胞系统或动物细胞系统来表达。在一些实施例中，本公开还教导了通过亲和标签或定制抗体纯化对蛋白质和或多肽的纯化。在其它实施例中，本公开还教导了针对多核苷酸进行化学合成的方法。
在一些实施例中，本领域技术人员将认识到，用于基因表达的病毒载体或质粒可以用于递送本文所公开的复合物。病毒样颗粒(VLP)可以用于包封核糖核蛋白复合物，并且本文所公开的经过纯化的核糖核蛋白复合物可以被纯化并通过电穿孔或注射来递送到细胞。
试剂盒
在一些实施例中，本公开提供了含有以上方法和组合物中所公开的元件中的任何一或多个元件的试剂盒。在一些实施例中，试剂盒包括模块化CRISPR DNA构建体和用于使用试剂盒的说明书以及任何必需的试剂或反应物。在一些实施例中，载体系统包括：(a)模块化CRISPR DNA构建体；(b)CRISPR复合物，包含CRISPR核酸内切酶蛋白和一或多个必需的靶引导RNA(或对所述项进行编码的序列)；以及任选地(c)插入DNA部分，如本申请的上文中所述。
元件可以单独或组合地提供并且可以设置在如小瓶、瓶子或试管等任何适合的容器或宿主细胞或质粒中。在一些实施例中，试剂盒包含一或多种语言例如多于一种语言的说明书。
在一些实施例中，试剂盒包括用于在利用本文所描述的元件中的一或多个元件的方法中使用的一或多种试剂(例如，经过纯化的Cpf1核酸内切酶)。试剂可以设置在任何适合的容器中。例如，试剂盒可以提供一或多种反应或储存缓冲液。试剂可以以可用于特定测定中的形式或以在使用前需要添加一或多种其它组分的形式(例如，以浓缩物或冻干的形式)提供。缓冲液可以是任何缓冲液，包含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缓冲液和其组合。在一些实施例中，缓冲液是碱性的。在一些实施例中，缓冲液的pH为约7到约10。在一些实施例中，试剂盒包括一或多种与crRNA序列相对应的寡核苷酸以用于插入到载体中以可操作地连接crRNA序列和调节元件。
实例
以下实例出于展示本公开的各种实施例的目的而给出并且不意味着以任何方式限制本公开。如权利要求的范围所定义的，本领域技术人员将想到涵盖在本公开的精神内的实例变化和其它用途。
实例1：一锅式体外模块化CRISPR克隆
本实例描述了在一锅式反应中通过将插入物从一种质粒转移到另一种质粒来生成质粒13001009086(SEQ ID NO:82)。参见图4。
两种质粒均携带侧接感兴趣区域的克隆标签(cTAG K[SEQ ID NO:78]/cTAG L[SEQ ID NO:79]和cTAG K'[SEQ ID NO:80]/cTAG L'[SEQ ID NO:81])。为了驱动对经过编辑的质粒的克隆反应，Cpf1间隔子在受体和供体质粒上处于相反朝向(分别为K/K'和L/L')。此由内向外/由外向内消化去除了最终产物中的Cpf1间隔子，从而消除了对期望的产物的重新切割(参见图4中的弯曲箭头，其描述了'485质粒中的由内向外消化和'784质粒中的由外向内消化)。Cas9间隔子保留，从而实现了在此位点处的迭代编辑。因此，大模块化构建体允许实现迭代编辑的快速单锅式反应方案。
Cpf1蛋白由金斯瑞公司(Genscript)合成，并且crRNA由新瑟果公司(Synthego)合成。对于一锅式切割/连接反应，将与crRNA(crRNA 1和crRNA 3)复合的Cpf1蛋白添加到含有ATP的缓冲液中的质粒(13000789485—SEQ ID NO:83和13000823784—SEQ ID NO:84)和DNA连接酶。这些组分在针对切割和连接进行优化的温度下循环。
将此反应转化到大肠杆菌中，并且对阳性克隆进行测序以确认新插入物的插入和Cpf1间隔子的丢失。
对于缺失，所使用的克隆标签中的Cpf1位点必须生成相容的突出端以允许质粒关闭。cTAG L'被设计成含有两个Cpf1间隔子，一个用于插入，其中突出端与cTAG K'不相容，而另一个用于缺失，其中突出端与cTAG K'相容。
实例2：体外模块化CRISPR克隆
本实例是为了展现CRISPR克隆的灵活性而设计的。作为初始步骤，从源载体pzHR039(SEQ ID No:100)和13000223370(SEQ ID No:101)分别产生针对卡那霉素或氯霉素抗性基因进行编码的几个抗性质粒。卡那霉素抗性质粒各自被设计成包含侧接GFP基因的各种Cpf1着陆位点(当被消化时，这些质粒产生“抗卡那霉素质粒主链”)。氯霉素抗性质粒各自被设计成包含侧接氯霉素抗性基因的各种Cpf1着陆位点(当被消化时，这些质粒产生“抗氯霉素插入物”)。本实例中使用的每个质粒的序列和载体图在表6中公开。
每个抗卡那霉素和氯霉素质粒最初分别用II型限制性酶KpnI-HF和PvuI-HF线性化(两者均可从NEB商购获得)。每个质粒上的KpnI和PvuI限制性位点的定位在图7-14提供的载体图中指出。线性化后，抗性质粒不再能够在细菌宿主系统中自复制。
然后将线性化抗性质粒与15μg(1.58μM最终浓度)Cpf1酶和下文描述的每个引导RNA 5μM共2μL(0.167μM最终浓度)的预先温育的混合物在60μL反应中混合，以形成活性CRISPR复合物。
本实例中使用的Cpf1酶可从IDT商购获得。Cpf1来源于氨基酸球菌Cpf1(AsCpf1)。将酶进一步修饰成包括1个N端核定位序列(NLS)和1个C端NLS以及3个N端FLAG标签和C端6-His标签。
本实例中使用的引导RNA是从IDT定制订购的。每个引导RNA被设计成靶向定位于线性化抗性质粒内的不同CRISPR着陆位点。在本实例中，切除了主链质粒的Cpf1着陆位点，但是在连接了插入物后恢复。表6提供了所使用的每个引导RNA的引导序列部分。因此，混合物中的CRISPR复合物被设计成从每个抗卡那霉素质粒中切割出GFP基因，以生成抗卡那霉素性质粒主链(参见图5，第二小图)。因此，混合物中的CRISPR复合物还被设计成从氯霉素抗性质粒中切割出氯霉素抗性基因，以生成抗氯霉素插入物(参见图5，第二小图)。类似地设计每个反应的抗卡那霉素质粒主链和抗氯霉素插入物以生成相容的突出端，所述相容的突出端将导致末端杂交以产生“双抗”卡那霉素和氯霉素质粒。
使包括Cpf1和引导RNA的线性化抗性质粒混合物在制造商推荐的Cpf1缓冲液中在37摄氏度下温育3小时。在琼脂糖凝胶上运行所选反应，使用标准的DNA提取试剂盒(齐莫研究公司(Zymo Research)试剂盒，根据制造商的说明书使用)对所得的片段进行纯化。经过纯化(对照)和未纯化(测试)。
使包括各自包含两个相容的Cpf1粘性末端的抗卡那霉素质粒主链和抗氯霉素插入物的DNA片段在有或没有T4 DNA连接酶(从NEB商购获得)的新反应中组合并转化为NEB10-B细胞(可从NEB商购获得)。将转化的细胞平板接种于用卡那霉素和氯霉素两者增强的培养基上，所述培养基被设计成防止不含功能性抗性质粒的任何细胞生长。
将单独菌落送去进行桑格尔(Sanger)测序以确认Cpf1克隆的接合。还使用表6中所述的引物通过PCR验证了回收的菌落。图5展示了上文所描述的一般实验性设计，例外的是如上文所描述的，质粒在Cpf1消化前被线性化。
表6：本实例2中使用的序列的列表
Figure BDA0002979126820000681
Figure BDA0002979126820000691
Figure BDA0002979126820000701
***用于SEQ ID NO:110-113的引导RNA的未加下划线部分是来自IDT的经过化学修饰的Alt-R RNA。与相应cTAG(即，M-P)的序列同源区域是加下划线的。
本实验的结果示出在表7和图6中。每种转化的反应编号沿着顶行示出，其中所使用的引导RNA沿着表7的左侧列列出。有和没有连接酶的相同Cpf1反应的比较示出了在连接酶的存在下转化子增加9.9倍，从而指示菌落生长是由于Cpf1消化后形成了卡那霉素和氯霉素双抗质粒。无连接酶反应是被设计成证实反应具有特异性的匹配的对照，而不是简单地由于存在污染水平的未消化的抗性质粒。
对十六个单独的菌落进行桑格尔测序以验证上游和下游克隆接合。在七个上游测序接合中的七个上游测序接合和九个下游接合中的八个下游接合中，来自具有T4 DNA连接酶的反应的Cpf1介导的克隆指示了忠实的消化和连接。
将反应71和72用未进行DNA凝胶纯化步骤的Cpf1消化的质粒进行转化。但是，在添加T4 DNA连接酶之前，根据供应商的说明书将Cpf1酶热灭活(反应72)。反应71和72表现出相同的连接酶依赖性。
表7：包括Cpf1编辑的载体的抗性转化子菌落
Figure BDA0002979126820000702
*用Cpf1消化后未进行DNA凝胶纯化的经过消化的DNA来转化板71和72。
PCT/US2017/042245(WO 2018/013990 A1，要求美国临时申请第62/362,909号的优先权)的公开内容以其全文并入本文。
实例3：使用大模块化设计通过限制性酶消化和连接进行的质粒组装
本实例描述了根据本公开的方法的模块化CRISPR载体的基因编辑。图15展示了本实例中描述的模块化CRISPR质粒13000444591的基因编辑。首先通过从先前构建的质粒中去除“填充物”插入DNA部分来制备质粒主链。通过用限制性酶ApaI和PvuI消化填充物部分的侧接克隆标签(cTAG)D(SEQ ID NO:68)和E(SEQ ID NO:69)来去除填充物插入DNA部分。通过凝胶电泳分离所得的片段，并且将与质粒主链相对应的期望的8.3kb片段从凝胶中切除并使用标准硅胶膜柱进行提取。
为了针对模块化CRISPR载体生成新插入物，使用通用的cTAG寡核苷酸tagD_FWD(SEQ ID NO:75)和tagE_REV(SEQ ID NO:76)对侧接有cTAG D和cTAG E的期望的插入DNA部分进行PCR扩增。所得的插入物含有侧接有cTAG D和cTAG E的GFP标志物基因。将所得的PCR片段用在cTAG D内切割的ApaI酶和在cTAG E内切割的PvuI酶消化。将消化的插入DNA部分使用标准硅胶膜柱纯化。
将纯化的模块化CRISPR载体主链和插入DNA部分组合到具有连接酶的单个反应中以生成环状质粒。(SEQ ID NO:77)中提供了所得的经过编辑的含GFP质粒13000444591的序列。
实例4：使用大模块化设计通过酵母同源重组进行的质粒组装
通过侧接有大模块化标签的PCR片段的酵母同源重组来组装质粒13000283399(SEQ ID NO:85)。用于组装的期望的构建体以这样的方式通过PCR进行扩增使得所述构建体侧接有特定大模块化标签。这些标签允许定向组装酿酒酵母中的片段，因为标签本身用作用于同源重组的重叠同源区域。具体地，如下通过PCR扩增侧接有大模块化标签的5个片段：标签A-片段1-标签B；标签B-片段2-标签C；标签C-片段3-标签D；标签D-片段4-标签E；以及标签E-片段5-标签F。将这些片段连同含有酵母复制起点和TRP营养缺陷型选择标志物以及一个末端处的标签A和另一个末端处的标签B的线性化组装载体转化为酿酒酵母。通过缺乏色氨酸的培养基中的酿酒酵母生长来选择环化组装质粒。将这些质粒回收并在大肠杆菌中扩增，并且通过测序来确认正确的构象。
实例5：棒状杆菌中的CRISPRi验证
在谷氨酸棒状杆菌的非典型种中测试本公开的CRISPRi方法。将CRISPRi系统的组分克隆到针对dCas9和引导RNA进行编码的单个测试载体中。将测试载体中的引导RNA设计成靶向与针对“辣椒”红色荧光蛋白(RFP)进行编码的基因可操作地连接的第二构建体的启动子区域。生成其中引导RNA是未靶向辣椒RFP的单独序列的第二对照载体。
将上文所描述的测试和对照载体转化到野生型(WT)谷氨酸棒杆菌菌株和包括上文所描述的启动子-RFP构建体的pcg2613-辣椒菌株以及缺少RFP构建体的WT菌株中(参见图17A中的实验设计)。将培养物在补充有2％阿拉伯糖的培养基中在30℃下生长48小时，以通过pBAD启动子诱导dCas9。然后记录针对7个生物复制物的活门控细胞的中值荧光。本实验的结果在图17B上示出。WT菌株均未表现出任何RFP荧光。含有对照CRISPRi质粒的pcg2613菌株表现出正常水平的RFP荧光。但是，测试CRISPRi质粒成功地敲低了RFP基因的表达。
本发明的另外的实施例
在以下编号的实施例中阐述了本公开设想的其它主题：
1.一种用于调节宿主细胞基因的表达的重组模块化CRISPR DNA构建体，所述构建体包括CRISPR多克隆位点，所述多克隆位点包括：
a)至少两个不同的克隆标签(cTAG)，其中每个cTAG包括：
i)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序(PAM)可操作地连接的原间隔子序列；其中所述经过验证的CRISPR着陆位点中的至少一个在所述模块化CRISPR DNA构建体中是独特的；以及
b)一或多个DNA插入部分；
i)其中所述不同的cTAG中的每个cTAG围绕所述一或多个DNA插入部分中的每个DNA插入部分分布在侧翼位置；
其中所述构建体进一步包括：
c)第一核酸，所述第一核酸对催化灭活CRISPR酶进行编码；
d)第二核酸，所述第二核酸对能够将(c)的所述催化灭活CRISPR酶募集到DNA靶位点的引导RNA进行编码。
1.1.根据实施例1所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPRDNA构建体包括第一复制起点。
1.2.根据实施例1到1.1中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体包括多于一个复制起点。
1.3.根据实施例1到1.2中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体包括第一复制起点和第二复制起点。
1.4.根据实施例1.1和1.3中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第一复制起点能够维持大肠杆菌(E.coli)中的质粒。
1.5.根据实施例1.1、1.3和1.4中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第二复制起点能够维持谷氨酸棒状杆菌(Corynebacterium glutamicum)中的质粒。
1.6.根据实施例1到1.4中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第一复制起点能够维持大肠杆菌中的质粒，第二复制起点能够维持酿酒酵母(Saccharomyces cerevisiae)中的质粒，并且第三复制起点能够维持谷氨酸棒状杆菌中的质粒。
1.7.根据实施例1到1.6中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体包括针对可选择标志物进行编码的插入部分。
1.8.根据实施例1到1.7中任一实施例所述的重组模块化CRISPR DNA构建体，其中至少一个复制起点包括在所述CRISPR多克隆位点内的插入部分内。
2.根据实施例1到1.8中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第一核酸包括在所述CRISPR多克隆位点内的插入部分内。
3.根据实施例2所述的重组模块化CRISPR DNA构建体，其中包括所述第一核酸的所述插入部分进一步包括可选择标志物。
4.根据实施例1到3中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第二核酸包括在所述CRISPR多克隆位点内的插入部分内。
5.根据实施例4所述的重组模块化CRISPR DNA构建体，其中包括所述第二核酸的所述插入部分进一步包括可选择标志物。
6.根据实施例1到1.8中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第一核酸和所述第二核酸各自包括在所述CRISPR多克隆位点内的其自身的插入部分内。
7.根据实施例6所述的重组模块化CRISPR DNA构建体，其中包括所述第一核酸和所述第二核酸的所述插入部分各自包括可选择标志物。
8.根据实施例5到7中任一实施例所述的重组模块化CRISPR DNA构建体，其中包括第一核酸的所述插入部分中包括的所述可选择标志物和包括所述第二核酸的所述插入部分中包括的所述可选择标志物是不同的。
9.根据实施例1到5中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第一核酸和所述第二核酸包括在所述CRISPR多克隆位点内的同一插入部分内。
10.根据实施例1到9中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第一核酸与启动子可操作地连接。
10.1根据实施例1到10中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第一核酸与终止子可操作地连接。
11.根据实施例1到10.1中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第二核酸与启动子可操作地连接。
11.1根据实施例1到11中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第二核酸与终止子可操作地连接。
12.根据实施例10到11.1中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述启动子是异源启动子。
12.1根据实施例10到12中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述启动子是组成型启动子。
12.2根据实施例10到12中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述启动子是诱导型启动子。
13.根据实施例1到12.2中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第一核酸对与转录激活蛋白翻译融合的催化灭活CRISPR酶进行编码。
13.1根据实施例1到12.2中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第一核酸对与转录灭活蛋白翻译融合的催化灭活CRISPR酶进行编码。
13.2根据实施例1到12.2中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第一核酸对与转录阻遏物翻译融合的催化灭活CRISPR酶进行编码。
14.根据实施例1到12.2中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述构建体进一步包括(e)第三核酸，所述第三核酸对在被表达时能够将自身与所述催化灭活CRISPR酶连接的转录激活蛋白进行编码。
14.1根据实施例1到12.2中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述构建体进一步包括(e)第三核酸，所述第三核酸对在被表达时能够将自身与所述催化灭活CRISPR酶连接的转录灭活蛋白进行编码。
14.2根据实施例1到12.2中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述构建体进一步包括(e)第三核酸，所述第三核酸对在被表达时能够将自身与所述催化灭活CRISPR酶连接的转录阻遏蛋白进行编码。
15.根据实施例14所述的重组模块化CRISPR DNA构建体，其中所述转录激活蛋白经由连接适配子或通过蛋白质-蛋白质相互作用将自身与所述催化灭活CRISPR酶连接。
15.1根据实施例14.1所述的重组模块化CRISPR DNA构建体，其中所述转录灭活蛋白经由连接适配子或通过蛋白质-蛋白质相互作用将自身与所述催化灭活CRISPR酶连接。
15.2根据实施例14.2所述的重组模块化CRISPR DNA构建体，其中所述转录阻遏蛋白经由连接适配子或通过蛋白质-蛋白质相互作用将自身与所述催化灭活CRISPR酶连接。
16.根据实施例1到12.2中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第二核酸对与能够将自身与转录激活蛋白连接的适配子可操作地连接的引导RNA进行编码。
16.1根据实施例1到12.2中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第二核酸对与能够将自身与转录灭活蛋白连接的适配子可操作地连接的引导RNA进行编码。
16.2根据实施例1到12.2中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述第二核酸对与能够将自身与转录阻遏蛋白连接的适配子可操作地连接的引导RNA进行编码。
17.根据实施例13、14、15和16中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述转录激活蛋白选自由以下组成的组：VP16、VP64和VP160、VPR。
17.1根据实施例13.1、14.1、15.1和16.1中任一实施例所述的重组模块化CRISPRDNA构建体，其中所述转录灭活蛋白选自由以下组成的组：Mxi1、Tbx3、KRAB、EnR和SID。
17.2根据实施例13.2、14.2、15.2和16.2中任一实施例所述的重组模块化CRISPRDNA构建体，其中所述转录阻遏蛋白选自由以下组成的组：Mxi1、Tbx3、KRAB、EnR和SID。
18.根据实施例1到17.2中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体是环状的。
19.根据实施例1到17.2中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体是线性的。
20.根据实施例1到17.2中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体被整合到生物体的基因组中。
21.根据实施例1到20中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述不同的cTAG中的至少一个包括至少两个经过验证的CRISPR着陆位点。
22.根据实施例1到21中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述CRISPR着陆位点中的至少一个是用于Cas9核酸内切酶。
23.根据实施例1到22中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述CRISPR着陆位点中的至少一个是用于Cpf1核酸内切酶。
24.根据实施例1到23中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述不同的cTAG中的至少一个包括罕见的(≥8个碱基长)限制性核酸内切酶位点。
24.1根据实施例1到23中任一实施例所述的重组模块化CRISPR DNA构建体，其中每个cTAG包括罕见的(≥8个碱基长)限制性核酸内切酶位点
25.根据实施例1到24.1中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述催化灭活CRISPR酶是突变的Cas9核酸内切酶。
26.根据实施例1到24.1中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述催化灭活CRISPR酶是突变的Cpf1核酸内切酶。
27.根据实施例1到24.1中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述催化灭活CRISPR酶选自表1的载体中包含的dCas9基因中。
28.根据实施例1到24.1中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述催化灭活CRISPR酶选自由以下组成的组：新凶手弗朗西丝菌(UniProtKB—A0Q7Q2(CPF1_FRATN))、毛螺菌科细菌(UniProtKB—A0A182DWE3(A0A182DWE3_9FIRM))和氨基酸球菌(UniProtKB—U2UMQ6(CPF1 ACISB)。
29.根据实施例1到24.1中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述催化灭活CRISPR酶是AsCpf1(D908A)。
30.根据实施例1到26中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述重组模块化CRISPR DNA构建体针对能够将(c)的所述催化灭活CRISPR酶募集到DNA靶位点的多于一个引导RNA进行编码。
31.根据实施例30所述的重组模块化CRISPR DNA构建体，其中所述引导RNA中的至少一个包括与所述构建体中编码的另一个引导RNA不同的序列。
32.根据实施例30或31所述的重组模块化CRISPR DNA构建体，其中所述引导RNA中的至少一个靶向与所述构建体中编码的另一个引导RNA不同的DNA靶位点序列。
33.根据实施例1到32中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述重组模块化CRISPR DNA构建体针对多于一种催化灭活CRISPR酶进行编码。
34.根据实施例33所述的重组模块化CRISPR DNA构建体，其中所述催化灭活CRISPR酶中的至少一种包括与所述构建体中编码的另一种催化灭活CRISPR酶不同的序列。
35.根据实施例1到34中任一实施例所述的插入部分，其中所述cTAG中的一或多个选自由SEQ ID NO:65-74、78-81和其组合组成的组。
35.5.一种宿主细胞，其包括根据实施例1到35中任一实施例所述的重组模块化CRISPR DNA构建体。
36.一种调节一或多个宿主细胞基因的表达的高通量方法，所述方法包括将根据实施例1到35中任一实施例所述的重组模块化CRISPR DNA构建体引入宿主细胞中的步骤；其中所述引导RNA的所述DNA靶位点位于宿主细胞基因组内。
37.根据实施例36所述的高通量方法，其中将所述重组模块化CRISPR DNA构建体的至少一个插入部分整合到宿主细胞的基因组中。
38.根据实施例36所述的高通量方法，其中所述重组模块化CRISPR DNA构建体作为染色体外DNA保留在宿主细胞中。
39.根据实施例36所述的高通量方法，其中将根据实施例10到12.2中任一实施例所述的重组模块化CRISPR DNA构建体引入宿主细胞中。
40.根据实施例39所述的高通量方法，其进一步包括使宿主细胞与能够增加所述诱导型启动子的表达的化合物接触的步骤。
41.一种用于筛选CRISPR酶变体的重组模块化CRISPR DNA构建体，所述构建体包括CRISPR多克隆位点，所述多克隆位点包括：
a)至少两个不同的克隆标签(cTAG)，其中每个cTAG包括：
i)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序(PAM)可操作地连接的原间隔子序列；其中所述经过验证的CRISPR着陆位点中的至少一个在所述模块化CRISPR DNA构建体中是独特的；以及
b)一或多个DNA插入部分；
i)其中所述不同的cTAG中的每个cTAG围绕所述一或多个DNA插入部分中的每个DNA插入部分分布在侧翼位置；
其中所述构建体进一步包括：
c)第一核酸，所述第一核酸对蛋白质进行编码；
d)第二核酸，所述第二核酸对能够与DNA靶位点结合的引导RNA进行编码。
41.1一种用于筛选CRISPR酶变体的重组模块化CRISPR DNA构建体，所述构建体包括CRISPR多克隆位点，所述多克隆位点包括：
a)至少两个不同的克隆标签(cTAG)，其中每个cTAG包括：
i)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序(PAM)可操作地连接的原间隔子序列；其中所述经过验证的CRISPR着陆位点中的至少一个在所述模块化CRISPR DNA构建体中是独特的；以及
b)一或多个DNA插入部分；
i)其中所述不同的cTAG中的每个cTAG围绕所述一或多个DNA插入部分中的每个DNA插入部分分布在侧翼位置；
其中所述构建体进一步包括：
c)第一核酸，所述第一核酸对CRISPR酶或疑似具有CRISPR功能的酶(“推定的CRISPR酶”)进行编码；
d)第二核酸，所述第二核酸对能够与DNA靶位点结合的引导RNA进行编码。
41.2.一种宿主细胞，其包括根据实施例41或41.1所述的重组模块化CRISPR DNA构建体。
42.一种筛选CRISPR酶变体的高通量方法，所述方法包括以下步骤：
a)将根据实施例41或41.1所述的重组模块化CRISPR DNA构建体引入宿主细胞中；其中所述引导RNA的所述DNA靶位点位于宿主细胞基因组内；以及
b)测量在所述DNA靶位点处发生的DNA切割的程度。
43.一种用于调节宿主细胞基因的表达或工程化宿主细胞的基因组的重组模块化CRISPR DNA构建体，所述构建体包括CRISPR多克隆位点，所述多克隆位点包括：
a)至少两个不同的克隆标签(cTAG)，其中每个cTAG包括：
i)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序(PAM)可操作地连接的原间隔子序列；其中所述经过验证的CRISPR着陆位点中的至少一个在所述模块化CRISPR DNA构建体中是独特的；以及
b)一或多个DNA插入部分；
i)其中所述不同的cTAG中的每个cTAG围绕所述一或多个DNA插入部分中的每个DNA插入部分分布在侧翼位置；以及
其中所述一或多个DNA插入部分包括针对CRISPR功能调节剂进行编码的的DNA。
44.根据实施例43所述的重组模块化CRISPR DNA构建体，其中包括针对CRISPR功能调节剂进行编码的所述DNA的所述插入部分进一步包括可选择标志物。
45.根据实施例43或44所述的重组模块化CRISPR DNA构建体，其中所述CRISPR功能调节剂选自由以下组成的组：复制起点、可选择标志物、可逆选择标志物、抗CRISPR蛋白、启动子、终止子、dCas9蛋白、dCpf1蛋白、条形码、Cas9蛋白、Cpf1蛋白、DNA供体和促进多重化基因组编辑的蛋白质。
46.一种宿主细胞，其包括根据实施例43到45中任一实施例所述的重组模块化CRISPR DNA构建体。
47.根据实施例46所述的宿主细胞，其中所述宿主细胞包括对催化活性CRISPR酶进行编码的核酸分子和能够将所述催化活性CRISPR酶募集到DNA靶位点的引导RNA。
48.根据实施例46所述的宿主细胞，其中所述宿主细胞包括对催化灭活CRISPR酶进行编码的核酸分子和能够将所述催化灭活CRISPR酶募集到DNA靶位点的引导RNA。
49.根据实施例48所述的宿主细胞，其中所述催化灭活CRISPR酶与转录激活蛋白翻译融合。
49.1根据实施例48所述的宿主细胞，其中所述催化灭活CRISPR酶与转录灭活蛋白翻译融合。
49.2根据实施例48所述的宿主细胞，其中所述催化灭活CRISPR酶与转录阻遏蛋白翻译融合。
50.根据实施例48所述的宿主细胞，其中所述宿主细胞进一步包括对转录激活蛋白进行编码的核酸分子，所述转录激活蛋白在表达时能够将自身与所述催化灭活CRISPR酶连接。
50.1根据实施例48所述的宿主细胞，其中所述宿主细胞进一步包括对转录灭活蛋白进行编码的核酸分子，所述转录灭活蛋白在表达时能够将自身与所述催化灭活CRISPR酶连接。
50.2根据实施例48所述的宿主细胞，其中所述宿主细胞进一步包括对转录阻遏蛋白进行编码的核酸分子，所述转录阻遏蛋白在表达时能够将其自身与所述催化灭活CRISPR酶连接。
51.根据实施例50所述的宿主细胞，其中所述转录激活蛋白经由连接适配子或通过蛋白质-蛋白质相互作用将自身与所述催化灭活CRISPR酶连接。
51.1根据实施例50.1所述的宿主细胞，其中所述转录灭活蛋白经由连接适配子或通过蛋白质-蛋白质相互作用将自身与所述催化灭活CRISPR酶连接。
51.2根据实施例50.2所述的宿主细胞，其中所述转录阻遏蛋白经由连接适配子或通过蛋白质-蛋白质相互作用将自身与所述催化灭活CRISPR酶连接。
52.根据实施例51所述的宿主细胞，其中所述引导RNA与能够将自身与转录激活蛋白连接的适配子可操作地连接。
52.1根据实施例51.1所述的宿主细胞，其中所述引导RNA与能够将自身与转录灭活蛋白连接的适配子可操作地连接。
52.2根据实施例51.3所述的宿主细胞，其中所述引导RNA与能够将自身与转录阻遏蛋白连接的适配子可操作地连接。
53.根据实施例49、51和52中任一实施例所述的宿主细胞，其中所述转录激活蛋白选自由以下组成的组：VP16、VP64和VP160、VPR。
53.1根据实施例49.1、51.1和52.1中任一实施例所述的宿主细胞，其中所述转录灭活蛋白选自由以下组成的组：Mxi1、Tbx3、KRAB、EnR和SID。
53.2根据实施例49.2、51.2和52.2中任一实施例所述的宿主细胞，其中所述转录激活蛋白选自由以下组成的组：Mxi1、Tbx3、KRAB、EnR和SID。
54.根据实施例43到45中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体是环状的。
55.根据实施例43到45中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体是线性的。
56.根据实施例43到45中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体被整合到生物体的基因组中。
57.根据实施例43到45和54到56中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述不同的cTAG中的至少一个包括至少两个经过验证的CRISPR着陆位点。
58.根据实施例43到45和54到57中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述CRISPR着陆位点中的至少一个是用于Cas9核酸内切酶。
59.根据实施例43到45和54到58中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述CRISPR着陆位点中的至少一个是用于Cpf1核酸内切酶。
60.根据实施例43到45和54到59中任一实施例所述的重组模块化CRISPR DNA构建体，其中所述不同的cTAG中的至少一个包括罕见的(≥8个碱基长)限制性核酸内切酶位点。
61.根据实施例47到53.2中任一实施例所述的宿主细胞，其中所述催化灭活CRISPR酶是突变的Cas9核酸内切酶。
62.根据实施例48到53.2中任一实施例所述的宿主细胞，其中所述催化灭活CRISPR酶是突变的Cpf1核酸内切酶。
63.根据实施例47到53.2和61到62中任一实施例所述的宿主细胞，其中所述宿主细胞包括多于一个核酸引导RNA。
64.根据实施例63所述的宿主细胞，其中所述引导RNA中的至少一个包括与另一个引导RNA不同的序列。
65.根据实施例64所述的宿主细胞，其中所述引导RNA中的至少一个靶向与另一个引导RNA不同的DNA靶位点序列。
66.根据实施例48到53.2和61到65中任一实施例所述的宿主细胞，其中所述宿主细胞包括多于一种催化灭活CRISPR酶。
67.根据实施例66所述的宿主细胞，其中所述催化灭活CRISPR酶中的至少一种包括与所述构建体中编码的另一种催化灭活CRISPR酶不同的序列。
68.根据实施例43到67中任一实施例所述的宿主细胞，其中所述cTAG中的一或多个选自由SEQ ID NO:65-74、78-81和其组合组成的组。
69.一种调节一或多个宿主细胞基因的表达的高通量方法，所述方法包括将根据实施例43到45和54到60中任一实施例所述的重组模块化CRISPR DNA构建体引入宿主细胞中的步骤；其中引导RNA的DNA靶位点位于宿主细胞基因组内。
70.根据实施例69所述的高通量方法，其中将所述重组模块化CRISPR DNA构建体的至少一个插入部分整合到所述宿主细胞的基因组中。
71.根据实施例69或70所述的高通量方法，其中所述插入部分调节CRISPR蛋白的功能。
72.根据实施例69或70所述的高通量方法，其中所述插入部分调节gRNA的功能。
73.根据实施例69或70所述的高通量方法，其中所述重组模块化CRISPR DNA构建体作为染色体外DNA保留在宿主细胞中。
74.一种用于筛选CRISPR酶变体的重组模块化CRISPR DNA构建体，所述构建体包括CRISPR多克隆位点，所述多克隆位点包括：
a)至少两个不同的克隆标签(cTAG)，其中每个cTAG包括：
i)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序(PAM)可操作地连接的原间隔子序列；其中所述经过验证的CRISPR着陆位点中的至少一个在所述模块化CRISPR DNA构建体中是独特的；以及
b)一或多个DNA插入部分；
i)其中所述不同的cTAG中的每个cTAG围绕所述一或多个DNA插入部分中的每个DNA插入部分分布在侧翼位置；
其中所述构建体进一步包括：
c)第一核酸，所述第一核酸对CRISPR酶或疑似具有CRISPR功能的酶(“推定的CRISPR酶”)进行编码；以及
d)第二核酸，所述第二核酸对能够与DNA靶位点结合的引导RNA进行编码。
75.一种在宿主细胞中筛选CRISPR活性的高通量方法，所述方法包括以下步骤：
a)将根据实施例43到45、54到60和74中任一实施例所述的重组模块化CRISPR DNA构建体引入所述宿主细胞中；其中引导RNA的所述DNA靶位点位于宿主细胞基因组内；以及
b)测量在所述DNA靶位点处发生的DNA切割的程度。
76.一种在宿主细胞中筛选CRISPRi活性的高通量方法，所述方法包括以下步骤：
a)将根据实施例43到45、54到60和74中任一实施例所述的重组模块化CRISPR DNA构建体引入所述宿主细胞中；其中引导RNA的所述DNA靶位点位于宿主细胞基因组内；以及
b)测量在所述DNA靶位点处发生的转录调节的程度。
*****
通过引用并入
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序列表
<110> 齐默尔根公司(Zymergen Inc.)
<120> CRISPRi在高通量代谢工程中的应用
<130> ZYMR-030/01WO 327574-2149
<150> US 62/764,672
<151> 2018-08-15
<160> 126
<170> PatentIn版本3.5
<210> 1
<211> 984
<212> PRT
<213> 空肠弯曲杆菌
<220>
<221> MISC_FEATURE
<222> (1)..(984)
<223> Cas9
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Lys Asn Leu Pro Thr Lys Lys Gln Lys Arg Ile Leu Asp Lys Asn Tyr
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Gln Ala Ile Phe Lys His Leu Arg Tyr Ser Lys Glu Asn Ala Glu Ser
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Ala Thr Phe Met Glu Leu Lys Ala Trp His Ala Ile Arg Lys Ala Leu
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Glu Asn Gln Gly Leu Lys Asp Thr Trp Gln Asp Leu Ala Lys Lys Pro
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Asp Leu Leu Asp Glu Ile Gly Thr Ala Phe Ser Leu Tyr Lys Thr Asp
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Glu Asp Ile Gln Gln Tyr Leu Thr Asn Lys Val Pro Asn Ser Val Ile
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Asn Ala Leu Leu Val Ser Leu Asn Phe Asp Lys Phe Ile Glu Leu Ser
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Leu Lys Ser Leu Arg Lys Ile Leu Pro Leu Met Glu Gln Gly Lys Arg
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Asn Gln Lys Thr Ser Gln Leu Leu Pro Ala Ile Pro Ala Gln Glu Ile
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Arg Asn Pro Val Val Leu Arg Thr Leu Ser Gln Ala Arg Lys Val Ile
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Asn Ala Ile Ile Arg Gln Tyr Gly Ser Pro Ala Arg Val His Ile Glu
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Thr Gly Arg Glu Leu Gly Lys Ser Phe Lys Glu Arg Arg Glu Ile Gln
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Lys Gln Gln Glu Asp Asn Arg Thr Lys Arg Glu Ser Ala Val Gln Lys
515 520 525
Phe Lys Glu Leu Phe Ser Asp Phe Ser Ser Glu Pro Lys Ser Lys Asp
530 535 540
Ile Leu Lys Phe Arg Leu Tyr Glu Gln Gln His Gly Lys Cys Leu Tyr
545 550 555 560
Ser Gly Lys Glu Ile Asn Ile His Arg Leu Asn Glu Lys Gly Tyr Val
565 570 575
Glu Ile Asp His Ala Leu Pro Phe Ser Arg Thr Trp Asp Asp Ser Phe
580 585 590
Asn Asn Lys Val Leu Val Leu Ala Ser Glu Asn Gln Asn Lys Gly Asn
595 600 605
Gln Thr Pro Tyr Glu Trp Leu Gln Gly Lys Ile Asn Ser Glu Arg Trp
610 615 620
Lys Asn Phe Val Ala Leu Val Leu Gly Ser Gln Cys Ser Ala Ala Lys
625 630 635 640
Lys Gln Arg Leu Leu Thr Gln Val Ile Asp Asp Asn Lys Phe Ile Asp
645 650 655
Arg Asn Leu Asn Asp Thr Arg Tyr Ile Ala Arg Phe Leu Ser Asn Tyr
660 665 670
Ile Gln Glu Asn Leu Leu Leu Val Gly Lys Asn Lys Lys Asn Val Phe
675 680 685
Thr Pro Asn Gly Gln Ile Thr Ala Leu Leu Arg Ser Arg Trp Gly Leu
690 695 700
Ile Lys Ala Arg Glu Asn Asn Asn Arg His His Ala Leu Asp Ala Ile
705 710 715 720
Val Val Ala Cys Ala Thr Pro Ser Met Gln Gln Lys Ile Thr Arg Phe
725 730 735
Ile Arg Phe Lys Glu Val His Pro Tyr Lys Ile Glu Asn Arg Tyr Glu
740 745 750
Met Val Asp Gln Glu Ser Gly Glu Ile Ile Ser Pro His Phe Pro Glu
755 760 765
Pro Trp Ala Tyr Phe Arg Gln Glu Val Asn Ile Arg Val Phe Asp Asn
770 775 780
His Pro Asp Thr Val Leu Lys Glu Met Leu Pro Asp Arg Pro Gln Ala
785 790 795 800
Asn His Gln Phe Val Gln Pro Leu Phe Val Ser Arg Ala Pro Thr Arg
805 810 815
Lys Met Ser Gly Gln Gly His Met Glu Thr Ile Lys Ser Ala Lys Arg
820 825 830
Leu Ala Glu Gly Ile Ser Val Leu Arg Ile Pro Leu Thr Gln Leu Lys
835 840 845
Pro Asn Leu Leu Glu Asn Met Val Asn Lys Glu Arg Glu Pro Ala Leu
850 855 860
Tyr Ala Gly Leu Lys Ala Arg Leu Ala Glu Phe Asn Gln Asp Pro Ala
865 870 875 880
Lys Ala Phe Ala Thr Pro Phe Tyr Lys Gln Gly Gly Gln Gln Val Lys
885 890 895
Ala Ile Arg Val Glu Gln Val Gln Lys Ser Gly Val Leu Val Arg Glu
900 905 910
Asn Asn Gly Val Ala Asp Asn Ala Ser Ile Val Arg Thr Asp Val Phe
915 920 925
Ile Lys Asn Asn Lys Phe Phe Leu Val Pro Ile Tyr Thr Trp Gln Val
930 935 940
Ala Lys Gly Ile Leu Pro Asn Lys Ala Ile Val Ala His Lys Asn Glu
945 950 955 960
Asp Glu Trp Glu Glu Met Asp Glu Gly Ala Lys Phe Lys Phe Ser Leu
965 970 975
Phe Pro Asn Asp Leu Val Glu Leu Lys Thr Lys Lys Glu Tyr Phe Phe
980 985 990
Gly Tyr Tyr Ile Gly Leu Asp Arg Ala Thr Gly Asn Ile Ser Leu Lys
995 1000 1005
Glu His Asp Gly Glu Ile Ser Lys Gly Lys Asp Gly Val Tyr Arg
1010 1015 1020
Val Gly Val Lys Leu Ala Leu Ser Phe Glu Lys Tyr Gln Val Asp
1025 1030 1035
Glu Leu Gly Lys Asn Arg Gln Ile Cys Arg Pro Gln Gln Arg Gln
1040 1045 1050
Pro Val Arg
1055
<210> 3
<211> 1368
<212> PRT
<213> 化脓链球菌
<220>
<221> MISC_FEATURE
<222> (1)..(1368)
<223> Cas9
<400> 3
Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val
1 5 10 15
Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe
20 25 30
Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile
35 40 45
Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu
50 55 60
Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys
65 70 75 80
Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser
85 90 95
Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys
100 105 110
His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr
115 120 125
His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp
130 135 140
Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His
145 150 155 160
Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro
165 170 175
Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr
180 185 190
Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala
195 200 205
Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn
210 215 220
Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn
225 230 235 240
Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe
245 250 255
Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp
260 265 270
Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp
275 280 285
Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp
290 295 300
Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser
305 310 315 320
Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys
325 330 335
Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe
340 345 350
Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser
355 360 365
Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp
370 375 380
Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg
385 390 395 400
Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu
405 410 415
Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe
420 425 430
Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile
435 440 445
Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp
450 455 460
Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu
465 470 475 480
Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr
485 490 495
Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser
500 505 510
Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys
515 520 525
Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln
530 535 540
Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr
545 550 555 560
Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp
565 570 575
Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly
580 585 590
Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp
595 600 605
Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr
610 615 620
Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala
625 630 635 640
His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr
645 650 655
Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp
660 665 670
Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe
675 680 685
Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe
690 695 700
Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu
705 710 715 720
His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly
725 730 735
Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly
740 745 750
Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln
755 760 765
Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile
770 775 780
Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro
785 790 795 800
Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu
805 810 815
Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg
820 825 830
Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys
835 840 845
Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg
850 855 860
Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys
865 870 875 880
Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys
885 890 895
Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp
900 905 910
Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr
915 920 925
Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp
930 935 940
Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser
945 950 955 960
Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg
965 970 975
Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val
980 985 990
Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe
995 1000 1005
Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala
1010 1015 1020
Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe
1025 1030 1035
Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala
1040 1045 1050
Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu
1055 1060 1065
Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val
1070 1075 1080
Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr
1085 1090 1095
Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys
1100 1105 1110
Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro
1115 1120 1125
Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser Val
1130 1135 1140
Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys
1145 1150 1155
Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser
1160 1165 1170
Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys
1175 1180 1185
Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu
1190 1195 1200
Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly
1205 1210 1215
Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val
1220 1225 1230
Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser
1235 1240 1245
Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys
1250 1255 1260
His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys
1265 1270 1275
Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala
1280 1285 1290
Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn
1295 1300 1305
Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala
1310 1315 1320
Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser
1325 1330 1335
Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr
1340 1345 1350
Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp
1355 1360 1365
<210> 4
<211> 1121
<212> PRT
<213> 嗜热链球菌
<220>
<221> MISC_FEATURE
<222> (1)..(1121)
<223> Cas9
<400> 4
Met Ser Asp Leu Val Leu Gly Leu Asp Ile Gly Ile Gly Ser Val Gly
1 5 10 15
Val Gly Ile Leu Asn Lys Val Thr Gly Glu Ile Ile His Lys Asn Ser
20 25 30
Arg Ile Phe Pro Ala Ala Gln Ala Glu Asn Asn Leu Val Arg Arg Thr
35 40 45
Asn Arg Gln Gly Arg Arg Leu Ala Arg Arg Lys Lys His Arg Arg Val
50 55 60
Arg Leu Asn Arg Leu Phe Glu Glu Ser Gly Leu Ile Thr Asp Phe Thr
65 70 75 80
Lys Ile Ser Ile Asn Leu Asn Pro Tyr Gln Leu Arg Val Lys Gly Leu
85 90 95
Thr Asp Glu Leu Ser Asn Glu Glu Leu Phe Ile Ala Leu Lys Asn Met
100 105 110
Val Lys His Arg Gly Ile Ser Tyr Leu Asp Asp Ala Ser Asp Asp Gly
115 120 125
Asn Ser Ser Val Gly Asp Tyr Ala Gln Ile Val Lys Glu Asn Ser Lys
130 135 140
Gln Leu Glu Thr Lys Thr Pro Gly Gln Ile Gln Leu Glu Arg Tyr Gln
145 150 155 160
Thr Tyr Gly Gln Leu Arg Gly Asp Phe Thr Val Glu Lys Asp Gly Lys
165 170 175
Lys His Arg Leu Ile Asn Val Phe Pro Thr Ser Ala Tyr Arg Ser Glu
180 185 190
Ala Leu Arg Ile Leu Gln Thr Gln Gln Glu Phe Asn Pro Gln Ile Thr
195 200 205
Asp Glu Phe Ile Asn Arg Tyr Leu Glu Ile Leu Thr Gly Lys Arg Lys
210 215 220
Tyr Tyr His Gly Pro Gly Asn Glu Lys Ser Arg Thr Asp Tyr Gly Arg
225 230 235 240
Tyr Arg Thr Ser Gly Glu Thr Leu Asp Asn Ile Phe Gly Ile Leu Ile
245 250 255
Gly Lys Cys Thr Phe Tyr Pro Asp Glu Phe Arg Ala Ala Lys Ala Ser
260 265 270
Tyr Thr Ala Gln Glu Phe Asn Leu Leu Asn Asp Leu Asn Asn Leu Thr
275 280 285
Val Pro Thr Glu Thr Lys Lys Leu Ser Lys Glu Gln Lys Asn Gln Ile
290 295 300
Ile Asn Tyr Val Lys Asn Glu Lys Ala Met Gly Pro Ala Lys Leu Phe
305 310 315 320
Lys Tyr Ile Ala Lys Leu Leu Ser Cys Asp Val Ala Asp Ile Lys Gly
325 330 335
Tyr Arg Ile Asp Lys Ser Gly Lys Ala Glu Ile His Thr Phe Glu Ala
340 345 350
Tyr Arg Lys Met Lys Thr Leu Glu Thr Leu Asp Ile Glu Gln Met Asp
355 360 365
Arg Glu Thr Leu Asp Lys Leu Ala Tyr Val Leu Thr Leu Asn Thr Glu
370 375 380
Arg Glu Gly Ile Gln Glu Ala Leu Glu His Glu Phe Ala Asp Gly Ser
385 390 395 400
Phe Ser Gln Lys Gln Val Asp Glu Leu Val Gln Phe Arg Lys Ala Asn
405 410 415
Ser Ser Ile Phe Gly Lys Gly Trp His Asn Phe Ser Val Lys Leu Met
420 425 430
Met Glu Leu Ile Pro Glu Leu Tyr Glu Thr Ser Glu Glu Gln Met Thr
435 440 445
Ile Leu Thr Arg Leu Gly Lys Gln Lys Thr Thr Ser Ser Ser Asn Lys
450 455 460
Thr Lys Tyr Ile Asp Glu Lys Leu Leu Thr Glu Glu Ile Tyr Asn Pro
465 470 475 480
Val Val Ala Lys Ser Val Arg Gln Ala Ile Lys Ile Val Asn Ala Ala
485 490 495
Ile Lys Glu Tyr Gly Asp Phe Asp Asn Ile Val Ile Glu Met Ala Arg
500 505 510
Glu Thr Asn Glu Asp Asp Glu Lys Lys Ala Ile Gln Lys Ile Gln Lys
515 520 525
Ala Asn Lys Asp Glu Lys Asp Ala Ala Met Leu Lys Ala Ala Asn Gln
530 535 540
Tyr Asn Gly Lys Ala Glu Leu Pro His Ser Val Phe His Gly His Lys
545 550 555 560
Gln Leu Ala Thr Lys Ile Arg Leu Trp His Gln Gln Gly Glu Arg Cys
565 570 575
Leu Tyr Thr Gly Lys Thr Ile Ser Ile His Asp Leu Ile Asn Asn Ser
580 585 590
Asn Gln Phe Glu Val Asp His Ile Leu Pro Leu Ser Ile Thr Phe Asp
595 600 605
Asp Ser Leu Ala Asn Lys Val Leu Val Tyr Ala Thr Ala Asn Gln Glu
610 615 620
Lys Gly Gln Arg Thr Pro Tyr Gln Ala Leu Asp Ser Met Asp Asp Ala
625 630 635 640
Trp Ser Phe Arg Glu Leu Lys Ala Phe Val Arg Glu Ser Lys Thr Leu
645 650 655
Ser Asn Lys Lys Lys Glu Tyr Leu Leu Thr Glu Glu Asp Ile Ser Lys
660 665 670
Phe Asp Val Arg Lys Lys Phe Ile Glu Arg Asn Leu Val Asp Thr Arg
675 680 685
Tyr Ala Ser Arg Val Val Leu Asn Ala Leu Gln Glu His Phe Arg Ala
690 695 700
His Lys Ile Asp Thr Lys Val Ser Val Val Arg Gly Gln Phe Thr Ser
705 710 715 720
Gln Leu Arg Arg His Trp Gly Ile Glu Lys Thr Arg Asp Thr Tyr His
725 730 735
His His Ala Val Asp Ala Leu Ile Ile Ala Ala Ser Ser Gln Leu Asn
740 745 750
Leu Trp Lys Lys Gln Lys Asn Thr Leu Val Ser Tyr Ser Glu Asp Gln
755 760 765
Leu Leu Asp Ile Glu Thr Gly Glu Leu Ile Ser Asp Asp Glu Tyr Lys
770 775 780
Glu Ser Val Phe Lys Ala Pro Tyr Gln His Phe Val Asp Thr Leu Lys
785 790 795 800
Ser Lys Glu Phe Glu Asp Ser Ile Leu Phe Ser Tyr Gln Val Asp Ser
805 810 815
Lys Phe Asn Arg Lys Ile Ser Asp Ala Thr Ile Tyr Ala Thr Arg Gln
820 825 830
Ala Lys Val Gly Lys Asp Lys Ala Asp Glu Thr Tyr Val Leu Gly Lys
835 840 845
Ile Lys Asp Ile Tyr Thr Gln Asp Gly Tyr Asp Ala Phe Met Lys Ile
850 855 860
Tyr Lys Lys Asp Lys Ser Lys Phe Leu Met Tyr Arg His Asp Pro Gln
865 870 875 880
Thr Phe Glu Lys Val Ile Glu Pro Ile Leu Glu Asn Tyr Pro Asn Lys
885 890 895
Gln Ile Asn Glu Lys Gly Lys Glu Val Pro Cys Asn Pro Phe Leu Lys
900 905 910
Tyr Lys Glu Glu His Gly Tyr Ile Arg Lys Tyr Ser Lys Lys Gly Asn
915 920 925
Gly Pro Glu Ile Lys Ser Leu Lys Tyr Tyr Asp Ser Lys Leu Gly Asn
930 935 940
His Ile Asp Ile Thr Pro Lys Asp Ser Asn Asn Lys Val Val Leu Gln
945 950 955 960
Ser Val Ser Pro Trp Arg Ala Asp Val Tyr Phe Asn Lys Thr Thr Gly
965 970 975
Lys Tyr Glu Ile Leu Gly Leu Lys Tyr Ala Asp Leu Gln Phe Glu Lys
980 985 990
Gly Thr Gly Thr Tyr Lys Ile Ser Gln Glu Lys Tyr Asn Asp Ile Lys
995 1000 1005
Lys Lys Glu Gly Val Asp Ser Asp Ser Glu Phe Lys Phe Thr Leu
1010 1015 1020
Tyr Lys Asn Asp Leu Leu Leu Val Lys Asp Thr Glu Thr Lys Glu
1025 1030 1035
Gln Gln Leu Phe Arg Phe Leu Ser Arg Thr Met Pro Lys Gln Lys
1040 1045 1050
His Tyr Val Glu Leu Lys Pro Tyr Asp Lys Gln Lys Phe Glu Gly
1055 1060 1065
Gly Glu Ala Leu Ile Lys Val Leu Gly Asn Val Ala Asn Ser Gly
1070 1075 1080
Gln Cys Lys Lys Gly Leu Gly Lys Ser Asn Ile Ser Ile Tyr Lys
1085 1090 1095
Val Arg Thr Asp Val Leu Gly Asn Gln His Ile Ile Lys Asn Glu
1100 1105 1110
Gly Asp Lys Pro Lys Leu Asp Phe
1115 1120
<210> 5
<211> 1082
<212> PRT
<213> 脑膜炎奈瑟氏菌
<220>
<221> MISC_FEATURE
<222> (1)..(1082)
<223> Cas9
<400> 5
Met Ala Ala Phe Lys Pro Asn Pro Ile Asn Tyr Ile Leu Gly Leu Asp
1 5 10 15
Ile Gly Ile Ala Ser Val Gly Trp Ala Met Val Glu Ile Asp Glu Asp
20 25 30
Glu Asn Pro Ile Cys Leu Ile Asp Leu Gly Val Arg Val Phe Glu Arg
35 40 45
Ala Glu Val Pro Lys Thr Gly Asp Ser Leu Ala Met Ala Arg Arg Leu
50 55 60
Ala Arg Ser Val Arg Arg Leu Thr Arg Arg Arg Ala His Arg Leu Leu
65 70 75 80
Arg Ala Arg Arg Leu Leu Lys Arg Glu Gly Val Leu Gln Ala Ala Asp
85 90 95
Phe Asp Glu Asn Gly Leu Ile Lys Ser Leu Pro Asn Thr Pro Trp Gln
100 105 110
Leu Arg Ala Ala Ala Leu Asp Arg Lys Leu Thr Pro Leu Glu Trp Ser
115 120 125
Ala Val Leu Leu His Leu Ile Lys His Arg Gly Tyr Leu Ser Gln Arg
130 135 140
Lys Asn Glu Gly Glu Thr Ala Asp Lys Glu Leu Gly Ala Leu Leu Lys
145 150 155 160
Gly Val Ala Asp Asn Ala His Ala Leu Gln Thr Gly Asp Phe Arg Thr
165 170 175
Pro Ala Glu Leu Ala Leu Asn Lys Phe Glu Lys Glu Ser Gly His Ile
180 185 190
Arg Asn Gln Arg Gly Asp Tyr Ser His Thr Phe Ser Arg Lys Asp Leu
195 200 205
Gln Ala Glu Leu Ile Leu Leu Phe Glu Lys Gln Lys Glu Phe Gly Asn
210 215 220
Pro His Val Ser Gly Gly Leu Lys Glu Gly Ile Glu Thr Leu Leu Met
225 230 235 240
Thr Gln Arg Pro Ala Leu Ser Gly Asp Ala Val Gln Lys Met Leu Gly
245 250 255
His Cys Thr Phe Glu Pro Ala Glu Pro Lys Ala Ala Lys Asn Thr Tyr
260 265 270
Thr Ala Glu Arg Phe Ile Trp Leu Thr Lys Leu Asn Asn Leu Arg Ile
275 280 285
Leu Glu Gln Gly Ser Glu Arg Pro Leu Thr Asp Thr Glu Arg Ala Thr
290 295 300
Leu Met Asp Glu Pro Tyr Arg Lys Ser Lys Leu Thr Tyr Ala Gln Ala
305 310 315 320
Arg Lys Leu Leu Gly Leu Glu Asp Thr Ala Phe Phe Lys Gly Leu Arg
325 330 335
Tyr Gly Lys Asp Asn Ala Glu Ala Ser Thr Leu Met Glu Met Lys Ala
340 345 350
Tyr His Ala Ile Ser Arg Ala Leu Glu Lys Glu Gly Leu Lys Asp Lys
355 360 365
Lys Ser Pro Leu Asn Leu Ser Pro Glu Leu Gln Asp Glu Ile Gly Thr
370 375 380
Ala Phe Ser Leu Phe Lys Thr Asp Glu Asp Ile Thr Gly Arg Leu Lys
385 390 395 400
Asp Arg Ile Gln Pro Glu Ile Leu Glu Ala Leu Leu Lys His Ile Ser
405 410 415
Phe Asp Lys Phe Val Gln Ile Ser Leu Lys Ala Leu Arg Arg Ile Val
420 425 430
Pro Leu Met Glu Gln Gly Lys Arg Tyr Asp Glu Ala Cys Ala Glu Ile
435 440 445
Tyr Gly Asp His Tyr Gly Lys Lys Asn Thr Glu Glu Lys Ile Tyr Leu
450 455 460
Pro Pro Ile Pro Ala Asp Glu Ile Arg Asn Pro Val Val Leu Arg Ala
465 470 475 480
Leu Ser Gln Ala Arg Lys Val Ile Asn Gly Val Val Arg Arg Tyr Gly
485 490 495
Ser Pro Ala Arg Ile His Ile Glu Thr Ala Arg Glu Val Gly Lys Ser
500 505 510
Phe Lys Asp Arg Lys Glu Ile Glu Lys Arg Gln Glu Glu Asn Arg Lys
515 520 525
Asp Arg Glu Lys Ala Ala Ala Lys Phe Arg Glu Tyr Phe Pro Asn Phe
530 535 540
Val Gly Glu Pro Lys Ser Lys Asp Ile Leu Lys Leu Arg Leu Tyr Glu
545 550 555 560
Gln Gln His Gly Lys Cys Leu Tyr Ser Gly Lys Glu Ile Asn Leu Gly
565 570 575
Arg Leu Asn Glu Lys Gly Tyr Val Glu Ile Asp His Ala Leu Pro Phe
580 585 590
Ser Arg Thr Trp Asp Asp Ser Phe Asn Asn Lys Val Leu Val Leu Gly
595 600 605
Ser Glu Asn Gln Asn Lys Gly Asn Gln Thr Pro Tyr Glu Tyr Phe Asn
610 615 620
Gly Lys Asp Asn Ser Arg Glu Trp Gln Glu Phe Lys Ala Arg Val Glu
625 630 635 640
Thr Ser Arg Phe Pro Arg Ser Lys Lys Gln Arg Ile Leu Leu Gln Lys
645 650 655
Phe Asp Glu Asp Gly Phe Lys Glu Arg Asn Leu Asn Asp Thr Arg Tyr
660 665 670
Val Asn Arg Phe Leu Cys Gln Phe Val Ala Asp Arg Met Arg Leu Thr
675 680 685
Gly Lys Gly Lys Lys Arg Val Phe Ala Ser Asn Gly Gln Ile Thr Asn
690 695 700
Leu Leu Arg Gly Phe Trp Gly Leu Arg Lys Val Arg Ala Glu Asn Asp
705 710 715 720
Arg His His Ala Leu Asp Ala Val Val Val Ala Cys Ser Thr Val Ala
725 730 735
Met Gln Gln Lys Ile Thr Arg Phe Val Arg Tyr Lys Glu Met Asn Ala
740 745 750
Phe Asp Gly Lys Thr Ile Asp Lys Glu Thr Gly Glu Val Leu His Gln
755 760 765
Lys Thr His Phe Pro Gln Pro Trp Glu Phe Phe Ala Gln Glu Val Met
770 775 780
Ile Arg Val Phe Gly Lys Pro Asp Gly Lys Pro Glu Phe Glu Glu Ala
785 790 795 800
Asp Thr Pro Glu Lys Leu Arg Thr Leu Leu Ala Glu Lys Leu Ser Ser
805 810 815
Arg Pro Glu Ala Val His Glu Tyr Val Thr Pro Leu Phe Val Ser Arg
820 825 830
Ala Pro Asn Arg Lys Met Ser Gly Gln Gly His Met Glu Thr Val Lys
835 840 845
Ser Ala Lys Arg Leu Asp Glu Gly Val Ser Val Leu Arg Val Pro Leu
850 855 860
Thr Gln Leu Lys Leu Lys Asp Leu Glu Lys Met Val Asn Arg Glu Arg
865 870 875 880
Glu Pro Lys Leu Tyr Glu Ala Leu Lys Ala Arg Leu Glu Ala His Lys
885 890 895
Asp Asp Pro Ala Lys Ala Phe Ala Glu Pro Phe Tyr Lys Tyr Asp Lys
900 905 910
Ala Gly Asn Arg Thr Gln Gln Val Lys Ala Val Arg Val Glu Gln Val
915 920 925
Gln Lys Thr Gly Val Trp Val Arg Asn His Asn Gly Ile Ala Asp Asn
930 935 940
Ala Thr Met Val Arg Val Asp Val Phe Glu Lys Gly Asp Lys Tyr Tyr
945 950 955 960
Leu Val Pro Ile Tyr Ser Trp Gln Val Ala Lys Gly Ile Leu Pro Asp
965 970 975
Arg Ala Val Val Gln Gly Lys Asp Glu Glu Asp Trp Gln Leu Ile Asp
980 985 990
Asp Ser Phe Asn Phe Lys Phe Ser Leu His Pro Asn Asp Leu Val Glu
995 1000 1005
Val Ile Thr Lys Lys Ala Arg Met Phe Gly Tyr Phe Ala Ser Cys
1010 1015 1020
His Arg Gly Thr Gly Asn Ile Asn Ile Arg Ile His Asp Leu Asp
1025 1030 1035
His Lys Ile Gly Lys Asn Gly Ile Leu Glu Gly Ile Gly Val Lys
1040 1045 1050
Thr Ala Leu Ser Phe Gln Lys Tyr Gln Ile Asp Glu Leu Gly Lys
1055 1060 1065
Glu Ile Arg Pro Cys Arg Leu Lys Lys Arg Pro Pro Val Arg
1070 1075 1080
<210> 6
<211> 1345
<212> PRT
<213> 变形链球菌
<220>
<221> MISC_FEATURE
<222> (1)..(1345)
<223> Cas9
<400> 6
Met Lys Lys Pro Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val
1 5 10 15
Gly Trp Ala Val Val Thr Asp Asp Tyr Lys Val Pro Ala Lys Lys Met
20 25 30
Lys Val Leu Gly Asn Thr Asp Lys Ser His Ile Glu Lys Asn Leu Leu
35 40 45
Gly Ala Leu Leu Phe Asp Ser Gly Asn Thr Ala Glu Asp Arg Arg Leu
50 55 60
Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Arg Asn Arg Ile Leu
65 70 75 80
Tyr Leu Gln Glu Ile Phe Ser Glu Glu Met Gly Lys Val Asp Asp Ser
85 90 95
Phe Phe His Arg Leu Glu Asp Ser Phe Leu Val Thr Glu Asp Lys Arg
100 105 110
Gly Glu Arg His Pro Ile Phe Gly Asn Leu Glu Glu Glu Val Lys Tyr
115 120 125
His Glu Asn Phe Pro Thr Ile Tyr His Leu Arg Gln Tyr Leu Ala Asp
130 135 140
Asn Pro Glu Lys Val Asp Leu Arg Leu Val Tyr Leu Ala Leu Ala His
145 150 155 160
Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Phe Asp Thr
165 170 175
Arg Asn Asn Asp Val Gln Arg Leu Phe Gln Glu Phe Leu Ala Val Tyr
180 185 190
Asp Asn Thr Phe Glu Asn Ser Ser Leu Gln Glu Gln Asn Val Gln Val
195 200 205
Glu Glu Ile Leu Thr Asp Lys Ile Ser Lys Ser Ala Lys Lys Asp Arg
210 215 220
Val Leu Lys Leu Phe Pro Asn Glu Lys Ser Asn Gly Arg Phe Ala Glu
225 230 235 240
Phe Leu Lys Leu Ile Val Gly Asn Gln Ala Asp Phe Lys Lys His Phe
245 250 255
Glu Leu Glu Glu Lys Ala Pro Leu Gln Phe Ser Lys Asp Thr Tyr Glu
260 265 270
Glu Glu Leu Glu Val Leu Leu Ala Gln Ile Gly Asp Asn Tyr Ala Glu
275 280 285
Leu Phe Leu Ser Ala Lys Lys Leu Tyr Asp Ser Ile Leu Leu Ser Gly
290 295 300
Ile Leu Thr Val Thr Asp Val Gly Thr Lys Ala Pro Leu Ser Ala Ser
305 310 315 320
Met Ile Gln Arg Tyr Asn Glu His Gln Met Asp Leu Ala Gln Leu Lys
325 330 335
Gln Phe Ile Arg Gln Lys Leu Ser Asp Lys Tyr Asn Glu Val Phe Ser
340 345 350
Asp Val Ser Lys Asp Gly Tyr Ala Gly Tyr Ile Asp Gly Lys Thr Asn
355 360 365
Gln Glu Ala Phe Tyr Lys Tyr Leu Lys Gly Leu Leu Asn Lys Ile Glu
370 375 380
Gly Ser Gly Tyr Phe Leu Asp Lys Ile Glu Arg Glu Asp Phe Leu Arg
385 390 395 400
Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu
405 410 415
Gln Glu Met Arg Ala Ile Ile Arg Arg Gln Ala Glu Phe Tyr Pro Phe
420 425 430
Leu Ala Asp Asn Gln Asp Arg Ile Glu Lys Leu Leu Thr Phe Arg Ile
435 440 445
Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Lys Ser Asp Phe Ala Trp
450 455 460
Leu Ser Arg Lys Ser Ala Asp Lys Ile Thr Pro Trp Asn Phe Asp Glu
465 470 475 480
Ile Val Asp Lys Glu Ser Ser Ala Glu Ala Phe Ile Asn Arg Met Thr
485 490 495
Asn Tyr Asp Leu Tyr Leu Pro Asn Gln Lys Val Leu Pro Lys His Ser
500 505 510
Leu Leu Tyr Glu Lys Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys
515 520 525
Tyr Lys Thr Glu Gln Gly Lys Thr Ala Phe Phe Asp Ala Asn Met Lys
530 535 540
Gln Glu Ile Phe Asp Gly Val Phe Lys Val Tyr Arg Lys Val Thr Lys
545 550 555 560
Asp Lys Leu Met Asp Phe Leu Glu Lys Glu Phe Asp Glu Phe Arg Ile
565 570 575
Val Asp Leu Thr Gly Leu Asp Lys Glu Asn Lys Val Phe Asn Ala Ser
580 585 590
Tyr Gly Thr Tyr His Asp Leu Cys Lys Ile Leu Asp Lys Asp Phe Leu
595 600 605
Asp Asn Ser Lys Asn Glu Lys Ile Leu Glu Asp Ile Val Leu Thr Leu
610 615 620
Thr Leu Phe Glu Asp Arg Glu Met Ile Arg Lys Arg Leu Glu Asn Tyr
625 630 635 640
Ser Asp Leu Leu Thr Lys Glu Gln Val Lys Lys Leu Glu Arg Arg His
645 650 655
Tyr Thr Gly Trp Gly Arg Leu Ser Ala Glu Leu Ile His Gly Ile Arg
660 665 670
Asn Lys Glu Ser Arg Lys Thr Ile Leu Asp Tyr Leu Ile Asp Asp Gly
675 680 685
Asn Ser Asn Arg Asn Phe Met Gln Leu Ile Asn Asp Asp Ala Leu Ser
690 695 700
Phe Lys Glu Glu Ile Ala Lys Ala Gln Val Ile Gly Glu Thr Asp Asn
705 710 715 720
Leu Asn Gln Val Val Ser Asp Ile Ala Gly Ser Pro Ala Ile Lys Lys
725 730 735
Gly Ile Leu Gln Ser Leu Lys Ile Val Asp Glu Leu Val Lys Ile Met
740 745 750
Gly His Gln Pro Glu Asn Ile Val Val Glu Met Ala Arg Glu Asn Gln
755 760 765
Phe Thr Asn Gln Gly Arg Arg Asn Ser Gln Gln Arg Leu Lys Gly Leu
770 775 780
Thr Asp Ser Ile Lys Glu Phe Gly Ser Gln Ile Leu Lys Glu His Pro
785 790 795 800
Val Glu Asn Ser Gln Leu Gln Asn Asp Arg Leu Phe Leu Tyr Tyr Leu
805 810 815
Gln Asn Gly Arg Asp Met Tyr Thr Gly Glu Glu Leu Asp Ile Asp Tyr
820 825 830
Leu Ser Gln Tyr Asp Ile Asp His Ile Ile Pro Gln Ala Phe Ile Lys
835 840 845
Asp Asn Ser Ile Asp Asn Arg Val Leu Thr Ser Ser Lys Glu Asn Arg
850 855 860
Gly Lys Ser Asp Asp Val Pro Ser Lys Asp Val Val Arg Lys Met Lys
865 870 875 880
Ser Tyr Trp Ser Lys Leu Leu Ser Ala Lys Leu Ile Thr Gln Arg Lys
885 890 895
Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Thr Asp Asp Asp
900 905 910
Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr
915 920 925
Lys His Val Ala Arg Ile Leu Asp Glu Arg Phe Asn Thr Glu Thr Asp
930 935 940
Glu Asn Asn Lys Lys Ile Arg Gln Val Lys Ile Val Thr Leu Lys Ser
945 950 955 960
Asn Leu Val Ser Asn Phe Arg Lys Glu Phe Glu Leu Tyr Lys Val Arg
965 970 975
Glu Ile Asn Asp Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val
980 985 990
Ile Gly Lys Ala Leu Leu Gly Val Tyr Pro Gln Leu Glu Pro Glu Phe
995 1000 1005
Val Tyr Gly Asp Tyr Pro His Phe His Gly His Lys Glu Asn Lys
1010 1015 1020
Ala Thr Ala Lys Lys Phe Phe Tyr Ser Asn Ile Met Asn Phe Phe
1025 1030 1035
Lys Lys Asp Asp Val Arg Thr Asp Lys Asn Gly Glu Ile Ile Trp
1040 1045 1050
Lys Lys Asp Glu His Ile Ser Asn Ile Lys Lys Val Leu Ser Tyr
1055 1060 1065
Pro Gln Val Asn Ile Val Lys Lys Val Glu Glu Gln Thr Gly Gly
1070 1075 1080
Phe Ser Lys Glu Ser Ile Leu Pro Lys Gly Asn Ser Asp Lys Leu
1085 1090 1095
Ile Pro Arg Lys Thr Lys Lys Phe Tyr Trp Asp Thr Lys Lys Tyr
1100 1105 1110
Gly Gly Phe Asp Ser Pro Ile Val Ala Tyr Ser Ile Leu Val Ile
1115 1120 1125
Ala Asp Ile Glu Lys Gly Lys Ser Lys Lys Leu Lys Thr Val Lys
1130 1135 1140
Ala Leu Val Gly Val Thr Ile Met Glu Lys Met Thr Phe Glu Arg
1145 1150 1155
Asp Pro Val Ala Phe Leu Glu Arg Lys Gly Tyr Arg Asn Val Gln
1160 1165 1170
Glu Glu Asn Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe Lys Leu
1175 1180 1185
Glu Asn Gly Arg Lys Arg Leu Leu Ala Ser Ala Arg Glu Leu Gln
1190 1195 1200
Lys Gly Asn Glu Ile Val Leu Pro Asn His Leu Gly Thr Leu Leu
1205 1210 1215
Tyr His Ala Lys Asn Ile His Lys Val Asp Glu Pro Lys His Leu
1220 1225 1230
Asp Tyr Val Asp Lys His Lys Asp Glu Phe Lys Glu Leu Leu Asp
1235 1240 1245
Val Val Ser Asn Phe Ser Lys Lys Tyr Thr Leu Ala Glu Gly Asn
1250 1255 1260
Leu Glu Lys Ile Lys Glu Leu Tyr Ala Gln Asn Asn Gly Glu Asp
1265 1270 1275
Leu Lys Glu Leu Ala Ser Ser Phe Ile Asn Leu Leu Thr Phe Thr
1280 1285 1290
Ala Ile Gly Ala Pro Ala Thr Phe Lys Phe Phe Asp Lys Asn Ile
1295 1300 1305
Asp Arg Lys Arg Tyr Thr Ser Thr Thr Glu Ile Leu Asn Ala Thr
1310 1315 1320
Leu Ile His Gln Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp
1325 1330 1335
Leu Asn Lys Leu Gly Gly Asp
1340 1345
<210> 7
<211> 1345
<212> PRT
<213> 土拉弗朗西斯菌亚种新凶手弗朗西斯菌U112
<220>
<221> MISC_FEATURE
<222> (1)..(1345)
<223> FnCpf1; pY004
<400> 7
Met Ser Ile Tyr Gln Glu Phe Val Asn Lys Tyr Ser Leu Ser Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Glu Asn Ile Lys
20 25 30
Ala Arg Gly Leu Ile Leu Asp Asp Glu Lys Arg Ala Lys Asp Tyr Lys
35 40 45
Lys Ala Lys Gln Ile Ile Asp Lys Tyr His Gln Phe Phe Ile Glu Glu
50 55 60
Ile Leu Ser Ser Val Cys Ile Ser Glu Asp Leu Leu Gln Asn Tyr Ser
65 70 75 80
Asp Val Tyr Phe Lys Leu Lys Lys Ser Asp Asp Asp Asn Leu Gln Lys
85 90 95
Asp Phe Lys Ser Ala Lys Asp Thr Ile Lys Lys Gln Ile Ser Glu Tyr
100 105 110
Ile Lys Asp Ser Glu Lys Phe Lys Asn Leu Phe Asn Gln Asn Leu Ile
115 120 125
Asp Ala Lys Lys Gly Gln Glu Ser Asp Leu Ile Leu Trp Leu Lys Gln
130 135 140
Ser Lys Asp Asn Gly Ile Glu Leu Phe Lys Ala Asn Ser Asp Ile Thr
145 150 155 160
Asp Ile Asp Glu Ala Leu Glu Ile Ile Lys Ser Phe Lys Gly Trp Thr
165 170 175
Thr Tyr Phe Lys Gly Phe His Glu Asn Arg Lys Asn Val Tyr Ser Ser
180 185 190
Asn Asp Ile Pro Thr Ser Ile Ile Tyr Arg Ile Val Asp Asp Asn Leu
195 200 205
Pro Lys Phe Leu Glu Asn Lys Ala Lys Tyr Glu Ser Leu Lys Asp Lys
210 215 220
Ala Pro Glu Ala Ile Asn Tyr Glu Gln Ile Lys Lys Asp Leu Ala Glu
225 230 235 240
Glu Leu Thr Phe Asp Ile Asp Tyr Lys Thr Ser Glu Val Asn Gln Arg
245 250 255
Val Phe Ser Leu Asp Glu Val Phe Glu Ile Ala Asn Phe Asn Asn Tyr
260 265 270
Leu Asn Gln Ser Gly Ile Thr Lys Phe Asn Thr Ile Ile Gly Gly Lys
275 280 285
Phe Val Asn Gly Glu Asn Thr Lys Arg Lys Gly Ile Asn Glu Tyr Ile
290 295 300
Asn Leu Tyr Ser Gln Gln Ile Asn Asp Lys Thr Leu Lys Lys Tyr Lys
305 310 315 320
Met Ser Val Leu Phe Lys Gln Ile Leu Ser Asp Thr Glu Ser Lys Ser
325 330 335
Phe Val Ile Asp Lys Leu Glu Asp Asp Ser Asp Val Val Thr Thr Met
340 345 350
Gln Ser Phe Tyr Glu Gln Ile Ala Ala Phe Lys Thr Val Glu Glu Lys
355 360 365
Ser Ile Lys Glu Thr Leu Ser Leu Leu Phe Asp Asp Leu Lys Ala Gln
370 375 380
Lys Leu Asp Leu Ser Lys Ile Tyr Phe Lys Asn Asp Lys Ser Leu Thr
385 390 395 400
Asp Leu Ser Gln Gln Val Phe Asp Asp Tyr Ser Val Ile Gly Thr Ala
405 410 415
Val Leu Glu Tyr Ile Thr Gln Gln Ile Ala Pro Lys Asn Leu Asp Asn
420 425 430
Pro Ser Lys Lys Glu Gln Glu Leu Ile Ala Lys Lys Thr Glu Lys Ala
435 440 445
Lys Tyr Leu Ser Leu Glu Thr Ile Lys Leu Ala Leu Glu Glu Phe Asn
450 455 460
Lys His Arg Asp Ile Asp Lys Gln Cys Arg Phe Glu Glu Ile Leu Ala
465 470 475 480
Asn Phe Ala Ala Ile Pro Met Ile Phe Asp Glu Ile Ala Gln Asn Lys
485 490 495
Asp Asn Leu Ala Gln Ile Ser Ile Lys Tyr Gln Asn Gln Gly Lys Lys
500 505 510
Asp Leu Leu Gln Ala Ser Ala Glu Asp Asp Val Lys Ala Ile Lys Asp
515 520 525
Leu Leu Asp Gln Thr Asn Asn Leu Leu His Lys Leu Lys Ile Phe His
530 535 540
Ile Ser Gln Ser Glu Asp Lys Ala Asn Ile Leu Asp Lys Asp Glu His
545 550 555 560
Phe Tyr Leu Val Phe Glu Glu Cys Tyr Phe Glu Leu Ala Asn Ile Val
565 570 575
Pro Leu Tyr Asn Lys Ile Arg Asn Tyr Ile Thr Gln Lys Pro Tyr Ser
580 585 590
Asp Glu Lys Phe Lys Leu Asn Phe Glu Asn Ser Thr Leu Ala Asn Gly
595 600 605
Trp Asp Lys Asn Lys Glu Pro Asp Asn Thr Ala Ile Leu Phe Ile Lys
610 615 620
Asp Asp Lys Tyr Tyr Leu Gly Val Met Asn Lys Lys Asn Asn Lys Ile
625 630 635 640
Phe Asp Asp Lys Ala Ile Lys Glu Asn Lys Gly Glu Gly Tyr Lys Lys
645 650 655
Ile Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro Lys Val
660 665 670
Phe Phe Ser Ala Lys Ser Ile Lys Phe Tyr Asn Pro Ser Glu Asp Ile
675 680 685
Leu Arg Ile Arg Asn His Ser Thr His Thr Lys Asn Gly Ser Pro Gln
690 695 700
Lys Gly Tyr Glu Lys Phe Glu Phe Asn Ile Glu Asp Cys Arg Lys Phe
705 710 715 720
Ile Asp Phe Tyr Lys Gln Ser Ile Ser Lys His Pro Glu Trp Lys Asp
725 730 735
Phe Gly Phe Arg Phe Ser Asp Thr Gln Arg Tyr Asn Ser Ile Asp Glu
740 745 750
Phe Tyr Arg Glu Val Glu Asn Gln Gly Tyr Lys Leu Thr Phe Glu Asn
755 760 765
Ile Ser Glu Ser Tyr Ile Asp Ser Val Val Asn Gln Gly Lys Leu Tyr
770 775 780
Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Ala Tyr Ser Lys Gly Arg
785 790 795 800
Pro Asn Leu His Thr Leu Tyr Trp Lys Ala Leu Phe Asp Glu Arg Asn
805 810 815
Leu Gln Asp Val Val Tyr Lys Leu Asn Gly Glu Ala Glu Leu Phe Tyr
820 825 830
Arg Lys Gln Ser Ile Pro Lys Lys Ile Thr His Pro Ala Lys Glu Ala
835 840 845
Ile Ala Asn Lys Asn Lys Asp Asn Pro Lys Lys Glu Ser Val Phe Glu
850 855 860
Tyr Asp Leu Ile Lys Asp Lys Arg Phe Thr Glu Asp Lys Phe Phe Phe
865 870 875 880
His Cys Pro Ile Thr Ile Asn Phe Lys Ser Ser Gly Ala Asn Lys Phe
885 890 895
Asn Asp Glu Ile Asn Leu Leu Leu Lys Glu Lys Ala Asn Asp Val His
900 905 910
Ile Leu Ser Ile Asp Arg Gly Glu Arg His Leu Ala Tyr Tyr Thr Leu
915 920 925
Val Asp Gly Lys Gly Asn Ile Ile Lys Gln Asp Thr Phe Asn Ile Ile
930 935 940
Gly Asn Asp Arg Met Lys Thr Asn Tyr His Asp Lys Leu Ala Ala Ile
945 950 955 960
Glu Lys Asp Arg Asp Ser Ala Arg Lys Asp Trp Lys Lys Ile Asn Asn
965 970 975
Ile Lys Glu Met Lys Glu Gly Tyr Leu Ser Gln Val Val His Glu Ile
980 985 990
Ala Lys Leu Val Ile Glu Tyr Asn Ala Ile Val Val Phe Glu Asp Leu
995 1000 1005
Asn Phe Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Val
1010 1015 1020
Tyr Gln Lys Leu Glu Lys Met Leu Ile Glu Lys Leu Asn Tyr Leu
1025 1030 1035
Val Phe Lys Asp Asn Glu Phe Asp Lys Thr Gly Gly Val Leu Arg
1040 1045 1050
Ala Tyr Gln Leu Thr Ala Pro Phe Glu Thr Phe Lys Lys Met Gly
1055 1060 1065
Lys Gln Thr Gly Ile Ile Tyr Tyr Val Pro Ala Gly Phe Thr Ser
1070 1075 1080
Lys Ile Cys Pro Val Thr Gly Phe Val Asn Gln Leu Tyr Pro Lys
1085 1090 1095
Tyr Glu Ser Val Ser Lys Ser Gln Glu Phe Phe Ser Lys Phe Asp
1100 1105 1110
Lys Ile Cys Tyr Asn Leu Asp Lys Gly Tyr Phe Glu Phe Ser Phe
1115 1120 1125
Asp Tyr Lys Asn Phe Gly Asp Lys Ala Ala Lys Gly Lys Trp Thr
1130 1135 1140
Ile Ala Ser Phe Gly Ser Arg Leu Ile Asn Phe Arg Asn Ser Asp
1145 1150 1155
Lys Asn His Asn Trp Asp Thr Arg Glu Val Tyr Pro Thr Lys Glu
1160 1165 1170
Leu Glu Lys Leu Leu Lys Asp Tyr Ser Ile Glu Tyr Gly His Gly
1175 1180 1185
Glu Cys Ile Lys Ala Ala Ile Cys Gly Glu Ser Asp Lys Lys Phe
1190 1195 1200
Phe Ala Lys Leu Thr Ser Val Leu Asn Thr Ile Leu Gln Met Arg
1205 1210 1215
Asn Ser Lys Thr Gly Thr Glu Leu Asp Tyr Leu Ile Ser Pro Val
1220 1225 1230
Ala Asp Val Asn Gly Asn Phe Phe Asp Ser Arg Gln Ala Pro Lys
1235 1240 1245
Asn Met Pro Gln Asp Ala Asp Ala Asn Gly Ala Tyr His Ile Gly
1250 1255 1260
Leu Lys Gly Leu Met Leu Leu Gly Arg Ile Lys Asn Asn Gln Glu
1265 1270 1275
Gly Lys Lys Leu Asn Leu Val Ile Lys Asn Glu Glu Tyr Phe Glu
1280 1285 1290
Phe Val Gln Asn Arg Asn Asn Lys Arg Pro Ala Ala Thr Lys Lys
1295 1300 1305
Ala Gly Gln Ala Lys Lys Lys Lys Gly Ser Tyr Pro Tyr Asp Val
1310 1315 1320
Pro Asp Tyr Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Tyr Pro
1325 1330 1335
Tyr Asp Val Pro Asp Tyr Ala
1340 1345
<210> 8
<211> 1278
<212> PRT
<213> 毛螺菌科细菌MC2017
<220>
<221> MISC_FEATURE
<222> (1)..(1278)
<223> Lb3Cpf1; pY005
<400> 8
Met Asp Tyr Gly Asn Gly Gln Phe Glu Arg Arg Ala Pro Leu Thr Lys
1 5 10 15
Thr Ile Thr Leu Arg Leu Lys Pro Ile Gly Glu Thr Arg Glu Thr Ile
20 25 30
Arg Glu Gln Lys Leu Leu Glu Gln Asp Ala Ala Phe Arg Lys Leu Val
35 40 45
Glu Thr Val Thr Pro Ile Val Asp Asp Cys Ile Arg Lys Ile Ala Asp
50 55 60
Asn Ala Leu Cys His Phe Gly Thr Glu Tyr Asp Phe Ser Cys Leu Gly
65 70 75 80
Asn Ala Ile Ser Lys Asn Asp Ser Lys Ala Ile Lys Lys Glu Thr Glu
85 90 95
Lys Val Glu Lys Leu Leu Ala Lys Val Leu Thr Glu Asn Leu Pro Asp
100 105 110
Gly Leu Arg Lys Val Asn Asp Ile Asn Ser Ala Ala Phe Ile Gln Asp
115 120 125
Thr Leu Thr Ser Phe Val Gln Asp Asp Ala Asp Lys Arg Val Leu Ile
130 135 140
Gln Glu Leu Lys Gly Lys Thr Val Leu Met Gln Arg Phe Leu Thr Thr
145 150 155 160
Arg Ile Thr Ala Leu Thr Val Trp Leu Pro Asp Arg Val Phe Glu Asn
165 170 175
Phe Asn Ile Phe Ile Glu Asn Ala Glu Lys Met Arg Ile Leu Leu Asp
180 185 190
Ser Pro Leu Asn Glu Lys Ile Met Lys Phe Asp Pro Asp Ala Glu Gln
195 200 205
Tyr Ala Ser Leu Glu Phe Tyr Gly Gln Cys Leu Ser Gln Lys Asp Ile
210 215 220
Asp Ser Tyr Asn Leu Ile Ile Ser Gly Ile Tyr Ala Asp Asp Glu Val
225 230 235 240
Lys Asn Pro Gly Ile Asn Glu Ile Val Lys Glu Tyr Asn Gln Gln Ile
245 250 255
Arg Gly Asp Lys Asp Glu Ser Pro Leu Pro Lys Leu Lys Lys Leu His
260 265 270
Lys Gln Ile Leu Met Pro Val Glu Lys Ala Phe Phe Val Arg Val Leu
275 280 285
Ser Asn Asp Ser Asp Ala Arg Ser Ile Leu Glu Lys Ile Leu Lys Asp
290 295 300
Thr Glu Met Leu Pro Ser Lys Ile Ile Glu Ala Met Lys Glu Ala Asp
305 310 315 320
Ala Gly Asp Ile Ala Val Tyr Gly Ser Arg Leu His Glu Leu Ser His
325 330 335
Val Ile Tyr Gly Asp His Gly Lys Leu Ser Gln Ile Ile Tyr Asp Lys
340 345 350
Glu Ser Lys Arg Ile Ser Glu Leu Met Glu Thr Leu Ser Pro Lys Glu
355 360 365
Arg Lys Glu Ser Lys Lys Arg Leu Glu Gly Leu Glu Glu His Ile Arg
370 375 380
Lys Ser Thr Tyr Thr Phe Asp Glu Leu Asn Arg Tyr Ala Glu Lys Asn
385 390 395 400
Val Met Ala Ala Tyr Ile Ala Ala Val Glu Glu Ser Cys Ala Glu Ile
405 410 415
Met Arg Lys Glu Lys Asp Leu Arg Thr Leu Leu Ser Lys Glu Asp Val
420 425 430
Lys Ile Arg Gly Asn Arg His Asn Thr Leu Ile Val Lys Asn Tyr Phe
435 440 445
Asn Ala Trp Thr Val Phe Arg Asn Leu Ile Arg Ile Leu Arg Arg Lys
450 455 460
Ser Glu Ala Glu Ile Asp Ser Asp Phe Tyr Asp Val Leu Asp Asp Ser
465 470 475 480
Val Glu Val Leu Ser Leu Thr Tyr Lys Gly Glu Asn Leu Cys Arg Ser
485 490 495
Tyr Ile Thr Lys Lys Ile Gly Ser Asp Leu Lys Pro Glu Ile Ala Thr
500 505 510
Tyr Gly Ser Ala Leu Arg Pro Asn Ser Arg Trp Trp Ser Pro Gly Glu
515 520 525
Lys Phe Asn Val Lys Phe His Thr Ile Val Arg Arg Asp Gly Arg Leu
530 535 540
Tyr Tyr Phe Ile Leu Pro Lys Gly Ala Lys Pro Val Glu Leu Glu Asp
545 550 555 560
Met Asp Gly Asp Ile Glu Cys Leu Gln Met Arg Lys Ile Pro Asn Pro
565 570 575
Thr Ile Phe Leu Pro Lys Leu Val Phe Lys Asp Pro Glu Ala Phe Phe
580 585 590
Arg Asp Asn Pro Glu Ala Asp Glu Phe Val Phe Leu Ser Gly Met Lys
595 600 605
Ala Pro Val Thr Ile Thr Arg Glu Thr Tyr Glu Ala Tyr Arg Tyr Lys
610 615 620
Leu Tyr Thr Val Gly Lys Leu Arg Asp Gly Glu Val Ser Glu Glu Glu
625 630 635 640
Tyr Lys Arg Ala Leu Leu Gln Val Leu Thr Ala Tyr Lys Glu Phe Leu
645 650 655
Glu Asn Arg Met Ile Tyr Ala Asp Leu Asn Phe Gly Phe Lys Asp Leu
660 665 670
Glu Glu Tyr Lys Asp Ser Ser Glu Phe Ile Lys Gln Val Glu Thr His
675 680 685
Asn Thr Phe Met Cys Trp Ala Lys Val Ser Ser Ser Gln Leu Asp Asp
690 695 700
Leu Val Lys Ser Gly Asn Gly Leu Leu Phe Glu Ile Trp Ser Glu Arg
705 710 715 720
Leu Glu Ser Tyr Tyr Lys Tyr Gly Asn Glu Lys Val Leu Arg Gly Tyr
725 730 735
Glu Gly Val Leu Leu Ser Ile Leu Lys Asp Glu Asn Leu Val Ser Met
740 745 750
Arg Thr Leu Leu Asn Ser Arg Pro Met Leu Val Tyr Arg Pro Lys Glu
755 760 765
Ser Ser Lys Pro Met Val Val His Arg Asp Gly Ser Arg Val Val Asp
770 775 780
Arg Phe Asp Lys Asp Gly Lys Tyr Ile Pro Pro Glu Val His Asp Glu
785 790 795 800
Leu Tyr Arg Phe Phe Asn Asn Leu Leu Ile Lys Glu Lys Leu Gly Glu
805 810 815
Lys Ala Arg Lys Ile Leu Asp Asn Lys Lys Val Lys Val Lys Val Leu
820 825 830
Glu Ser Glu Arg Val Lys Trp Ser Lys Phe Tyr Asp Glu Gln Phe Ala
835 840 845
Val Thr Phe Ser Val Lys Lys Asn Ala Asp Cys Leu Asp Thr Thr Lys
850 855 860
Asp Leu Asn Ala Glu Val Met Glu Gln Tyr Ser Glu Ser Asn Arg Leu
865 870 875 880
Ile Leu Ile Arg Asn Thr Thr Asp Ile Leu Tyr Tyr Leu Val Leu Asp
885 890 895
Lys Asn Gly Lys Val Leu Lys Gln Arg Ser Leu Asn Ile Ile Asn Asp
900 905 910
Gly Ala Arg Asp Val Asp Trp Lys Glu Arg Phe Arg Gln Val Thr Lys
915 920 925
Asp Arg Asn Glu Gly Tyr Asn Glu Trp Asp Tyr Ser Arg Thr Ser Asn
930 935 940
Asp Leu Lys Glu Val Tyr Leu Asn Tyr Ala Leu Lys Glu Ile Ala Glu
945 950 955 960
Ala Val Ile Glu Tyr Asn Ala Ile Leu Ile Ile Glu Lys Met Ser Asn
965 970 975
Ala Phe Lys Asp Lys Tyr Ser Phe Leu Asp Asp Val Thr Phe Lys Gly
980 985 990
Phe Glu Thr Lys Leu Leu Ala Lys Leu Ser Asp Leu His Phe Arg Gly
995 1000 1005
Ile Lys Asp Gly Glu Pro Cys Ser Phe Thr Asn Pro Leu Gln Leu
1010 1015 1020
Cys Gln Asn Asp Ser Asn Lys Ile Leu Gln Asp Gly Val Ile Phe
1025 1030 1035
Met Val Pro Asn Ser Met Thr Arg Ser Leu Asp Pro Asp Thr Gly
1040 1045 1050
Phe Ile Phe Ala Ile Asn Asp His Asn Ile Arg Thr Lys Lys Ala
1055 1060 1065
Lys Leu Asn Phe Leu Ser Lys Phe Asp Gln Leu Lys Val Ser Ser
1070 1075 1080
Glu Gly Cys Leu Ile Met Lys Tyr Ser Gly Asp Ser Leu Pro Thr
1085 1090 1095
His Asn Thr Asp Asn Arg Val Trp Asn Cys Cys Cys Asn His Pro
1100 1105 1110
Ile Thr Asn Tyr Asp Arg Glu Thr Lys Lys Val Glu Phe Ile Glu
1115 1120 1125
Glu Pro Val Glu Glu Leu Ser Arg Val Leu Glu Glu Asn Gly Ile
1130 1135 1140
Glu Thr Asp Thr Glu Leu Asn Lys Leu Asn Glu Arg Glu Asn Val
1145 1150 1155
Pro Gly Lys Val Val Asp Ala Ile Tyr Ser Leu Val Leu Asn Tyr
1160 1165 1170
Leu Arg Gly Thr Val Ser Gly Val Ala Gly Gln Arg Ala Val Tyr
1175 1180 1185
Tyr Ser Pro Val Thr Gly Lys Lys Tyr Asp Ile Ser Phe Ile Gln
1190 1195 1200
Ala Met Asn Leu Asn Arg Lys Cys Asp Tyr Tyr Arg Ile Gly Ser
1205 1210 1215
Lys Glu Arg Gly Glu Trp Thr Asp Phe Val Ala Gln Leu Ile Asn
1220 1225 1230
Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys
1235 1240 1245
Lys Gly Ser Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Tyr Pro Tyr
1250 1255 1260
Asp Val Pro Asp Tyr Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
1265 1270 1275
<210> 9
<211> 1345
<212> PRT
<213> 瘤胃产氢丁酸弧菌
<220>
<221> MISC_FEATURE
<222> (1)..(1345)
<223> BpCpf1; pY006
<400> 9
Met Ser Ile Tyr Gln Glu Phe Val Asn Lys Tyr Ser Leu Ser Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Glu Asn Ile Lys
20 25 30
Ala Arg Gly Leu Ile Leu Asp Asp Glu Lys Arg Ala Lys Asp Tyr Lys
35 40 45
Lys Ala Lys Gln Ile Ile Asp Lys Tyr His Gln Phe Phe Ile Glu Glu
50 55 60
Ile Leu Ser Ser Val Cys Ile Ser Glu Asp Leu Leu Gln Asn Tyr Ser
65 70 75 80
Asp Val Tyr Phe Lys Leu Lys Lys Ser Asp Asp Asp Asn Leu Gln Lys
85 90 95
Asp Phe Lys Ser Ala Lys Asp Thr Ile Lys Lys Gln Ile Ser Glu Tyr
100 105 110
Ile Lys Asp Ser Glu Lys Phe Lys Asn Leu Phe Asn Gln Asn Leu Ile
115 120 125
Asp Ala Lys Lys Gly Gln Glu Ser Asp Leu Ile Leu Trp Leu Lys Gln
130 135 140
Ser Lys Asp Asn Gly Ile Glu Leu Phe Lys Ala Asn Ser Asp Ile Thr
145 150 155 160
Asp Ile Asp Glu Ala Leu Glu Ile Ile Lys Ser Phe Lys Gly Trp Thr
165 170 175
Thr Tyr Phe Lys Gly Phe His Glu Asn Arg Lys Asn Val Tyr Ser Ser
180 185 190
Asn Asp Ile Pro Thr Ser Ile Ile Tyr Arg Ile Val Asp Asp Asn Leu
195 200 205
Pro Lys Phe Leu Glu Asn Lys Ala Lys Tyr Glu Ser Leu Lys Asp Lys
210 215 220
Ala Pro Glu Ala Ile Asn Tyr Glu Gln Ile Lys Lys Asp Leu Ala Glu
225 230 235 240
Glu Leu Thr Phe Asp Ile Asp Tyr Lys Thr Ser Glu Val Asn Gln Arg
245 250 255
Val Phe Ser Leu Asp Glu Val Phe Glu Ile Ala Asn Phe Asn Asn Tyr
260 265 270
Leu Asn Gln Ser Gly Ile Thr Lys Phe Asn Thr Ile Ile Gly Gly Lys
275 280 285
Phe Val Asn Gly Glu Asn Thr Lys Arg Lys Gly Ile Asn Glu Tyr Ile
290 295 300
Asn Leu Tyr Ser Gln Gln Ile Asn Asp Lys Thr Leu Lys Lys Tyr Lys
305 310 315 320
Met Ser Val Leu Phe Lys Gln Ile Leu Ser Asp Thr Glu Ser Lys Ser
325 330 335
Phe Val Ile Asp Lys Leu Glu Asp Asp Ser Asp Val Val Thr Thr Met
340 345 350
Gln Ser Phe Tyr Glu Gln Ile Ala Ala Phe Lys Thr Val Glu Glu Lys
355 360 365
Ser Ile Lys Glu Thr Leu Ser Leu Leu Phe Asp Asp Leu Lys Ala Gln
370 375 380
Lys Leu Asp Leu Ser Lys Ile Tyr Phe Lys Asn Asp Lys Ser Leu Thr
385 390 395 400
Asp Leu Ser Gln Gln Val Phe Asp Asp Tyr Ser Val Ile Gly Thr Ala
405 410 415
Val Leu Glu Tyr Ile Thr Gln Gln Ile Ala Pro Lys Asn Leu Asp Asn
420 425 430
Pro Ser Lys Lys Glu Gln Glu Leu Ile Ala Lys Lys Thr Glu Lys Ala
435 440 445
Lys Tyr Leu Ser Leu Glu Thr Ile Lys Leu Ala Leu Glu Glu Phe Asn
450 455 460
Lys His Arg Asp Ile Asp Lys Gln Cys Arg Phe Glu Glu Ile Leu Ala
465 470 475 480
Asn Phe Ala Ala Ile Pro Met Ile Phe Asp Glu Ile Ala Gln Asn Lys
485 490 495
Asp Asn Leu Ala Gln Ile Ser Ile Lys Tyr Gln Asn Gln Gly Lys Lys
500 505 510
Asp Leu Leu Gln Ala Ser Ala Glu Asp Asp Val Lys Ala Ile Lys Asp
515 520 525
Leu Leu Asp Gln Thr Asn Asn Leu Leu His Lys Leu Lys Ile Phe His
530 535 540
Ile Ser Gln Ser Glu Asp Lys Ala Asn Ile Leu Asp Lys Asp Glu His
545 550 555 560
Phe Tyr Leu Val Phe Glu Glu Cys Tyr Phe Glu Leu Ala Asn Ile Val
565 570 575
Pro Leu Tyr Asn Lys Ile Arg Asn Tyr Ile Thr Gln Lys Pro Tyr Ser
580 585 590
Asp Glu Lys Phe Lys Leu Asn Phe Glu Asn Ser Thr Leu Ala Asn Gly
595 600 605
Trp Asp Lys Asn Lys Glu Pro Asp Asn Thr Ala Ile Leu Phe Ile Lys
610 615 620
Asp Asp Lys Tyr Tyr Leu Gly Val Met Asn Lys Lys Asn Asn Lys Ile
625 630 635 640
Phe Asp Asp Lys Ala Ile Lys Glu Asn Lys Gly Glu Gly Tyr Lys Lys
645 650 655
Ile Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro Lys Val
660 665 670
Phe Phe Ser Ala Lys Ser Ile Lys Phe Tyr Asn Pro Ser Glu Asp Ile
675 680 685
Leu Arg Ile Arg Asn His Ser Thr His Thr Lys Asn Gly Ser Pro Gln
690 695 700
Lys Gly Tyr Glu Lys Phe Glu Phe Asn Ile Glu Asp Cys Arg Lys Phe
705 710 715 720
Ile Asp Phe Tyr Lys Gln Ser Ile Ser Lys His Pro Glu Trp Lys Asp
725 730 735
Phe Gly Phe Arg Phe Ser Asp Thr Gln Arg Tyr Asn Ser Ile Asp Glu
740 745 750
Phe Tyr Arg Glu Val Glu Asn Gln Gly Tyr Lys Leu Thr Phe Glu Asn
755 760 765
Ile Ser Glu Ser Tyr Ile Asp Ser Val Val Asn Gln Gly Lys Leu Tyr
770 775 780
Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Ala Tyr Ser Lys Gly Arg
785 790 795 800
Pro Asn Leu His Thr Leu Tyr Trp Lys Ala Leu Phe Asp Glu Arg Asn
805 810 815
Leu Gln Asp Val Val Tyr Lys Leu Asn Gly Glu Ala Glu Leu Phe Tyr
820 825 830
Arg Lys Gln Ser Ile Pro Lys Lys Ile Thr His Pro Ala Lys Glu Ala
835 840 845
Ile Ala Asn Lys Asn Lys Asp Asn Pro Lys Lys Glu Ser Val Phe Glu
850 855 860
Tyr Asp Leu Ile Lys Asp Lys Arg Phe Thr Glu Asp Lys Phe Phe Phe
865 870 875 880
His Cys Pro Ile Thr Ile Asn Phe Lys Ser Ser Gly Ala Asn Lys Phe
885 890 895
Asn Asp Glu Ile Asn Leu Leu Leu Lys Glu Lys Ala Asn Asp Val His
900 905 910
Ile Leu Ser Ile Asp Arg Gly Glu Arg His Leu Ala Tyr Tyr Thr Leu
915 920 925
Val Asp Gly Lys Gly Asn Ile Ile Lys Gln Asp Thr Phe Asn Ile Ile
930 935 940
Gly Asn Asp Arg Met Lys Thr Asn Tyr His Asp Lys Leu Ala Ala Ile
945 950 955 960
Glu Lys Asp Arg Asp Ser Ala Arg Lys Asp Trp Lys Lys Ile Asn Asn
965 970 975
Ile Lys Glu Met Lys Glu Gly Tyr Leu Ser Gln Val Val His Glu Ile
980 985 990
Ala Lys Leu Val Ile Glu Tyr Asn Ala Ile Val Val Phe Glu Asp Leu
995 1000 1005
Asn Phe Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Val
1010 1015 1020
Tyr Gln Lys Leu Glu Lys Met Leu Ile Glu Lys Leu Asn Tyr Leu
1025 1030 1035
Val Phe Lys Asp Asn Glu Phe Asp Lys Thr Gly Gly Val Leu Arg
1040 1045 1050
Ala Tyr Gln Leu Thr Ala Pro Phe Glu Thr Phe Lys Lys Met Gly
1055 1060 1065
Lys Gln Thr Gly Ile Ile Tyr Tyr Val Pro Ala Gly Phe Thr Ser
1070 1075 1080
Lys Ile Cys Pro Val Thr Gly Phe Val Asn Gln Leu Tyr Pro Lys
1085 1090 1095
Tyr Glu Ser Val Ser Lys Ser Gln Glu Phe Phe Ser Lys Phe Asp
1100 1105 1110
Lys Ile Cys Tyr Asn Leu Asp Lys Gly Tyr Phe Glu Phe Ser Phe
1115 1120 1125
Asp Tyr Lys Asn Phe Gly Asp Lys Ala Ala Lys Gly Lys Trp Thr
1130 1135 1140
Ile Ala Ser Phe Gly Ser Arg Leu Ile Asn Phe Arg Asn Ser Asp
1145 1150 1155
Lys Asn His Asn Trp Asp Thr Arg Glu Val Tyr Pro Thr Lys Glu
1160 1165 1170
Leu Glu Lys Leu Leu Lys Asp Tyr Ser Ile Glu Tyr Gly His Gly
1175 1180 1185
Glu Cys Ile Lys Ala Ala Ile Cys Gly Glu Ser Asp Lys Lys Phe
1190 1195 1200
Phe Ala Lys Leu Thr Ser Val Leu Asn Thr Ile Leu Gln Met Arg
1205 1210 1215
Asn Ser Lys Thr Gly Thr Glu Leu Asp Tyr Leu Ile Ser Pro Val
1220 1225 1230
Ala Asp Val Asn Gly Asn Phe Phe Asp Ser Arg Gln Ala Pro Lys
1235 1240 1245
Asn Met Pro Gln Asp Ala Asp Ala Asn Gly Ala Tyr His Ile Gly
1250 1255 1260
Leu Lys Gly Leu Met Leu Leu Gly Arg Ile Lys Asn Asn Gln Glu
1265 1270 1275
Gly Lys Lys Leu Asn Leu Val Ile Lys Asn Glu Glu Tyr Phe Glu
1280 1285 1290
Phe Val Gln Asn Arg Asn Asn Lys Arg Pro Ala Ala Thr Lys Lys
1295 1300 1305
Ala Gly Gln Ala Lys Lys Lys Lys Gly Ser Tyr Pro Tyr Asp Val
1310 1315 1320
Pro Asp Tyr Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Tyr Pro
1325 1330 1335
Tyr Asp Val Pro Asp Tyr Ala
1340 1345
<210> 10
<211> 1522
<212> PRT
<213> 异域菌门细菌GW2011_GWA_33_10
<220>
<221> MISC_FEATURE
<222> (1)..(1522)
<223> PeCpf1; pY007
<400> 10
Met Ser Asn Phe Phe Lys Asn Phe Thr Asn Leu Tyr Glu Leu Ser Lys
1 5 10 15
Thr Leu Arg Phe Glu Leu Lys Pro Val Gly Asp Thr Leu Thr Asn Met
20 25 30
Lys Asp His Leu Glu Tyr Asp Glu Lys Leu Gln Thr Phe Leu Lys Asp
35 40 45
Gln Asn Ile Asp Asp Ala Tyr Gln Ala Leu Lys Pro Gln Phe Asp Glu
50 55 60
Ile His Glu Glu Phe Ile Thr Asp Ser Leu Glu Ser Lys Lys Ala Lys
65 70 75 80
Glu Ile Asp Phe Ser Glu Tyr Leu Asp Leu Phe Gln Glu Lys Lys Glu
85 90 95
Leu Asn Asp Ser Glu Lys Lys Leu Arg Asn Lys Ile Gly Glu Thr Phe
100 105 110
Asn Lys Ala Gly Glu Lys Trp Lys Lys Glu Lys Tyr Pro Gln Tyr Glu
115 120 125
Trp Lys Lys Gly Ser Lys Ile Ala Asn Gly Ala Asp Ile Leu Ser Cys
130 135 140
Gln Asp Met Leu Gln Phe Ile Lys Tyr Lys Asn Pro Glu Asp Glu Lys
145 150 155 160
Ile Lys Asn Tyr Ile Asp Asp Thr Leu Lys Gly Phe Phe Thr Tyr Phe
165 170 175
Gly Gly Phe Asn Gln Asn Arg Ala Asn Tyr Tyr Glu Thr Lys Lys Glu
180 185 190
Ala Ser Thr Ala Val Ala Thr Arg Ile Val His Glu Asn Leu Pro Lys
195 200 205
Phe Cys Asp Asn Val Ile Gln Phe Lys His Ile Ile Lys Arg Lys Lys
210 215 220
Asp Gly Thr Val Glu Lys Thr Glu Arg Lys Thr Glu Tyr Leu Asn Ala
225 230 235 240
Tyr Gln Tyr Leu Lys Asn Asn Asn Lys Ile Thr Gln Ile Lys Asp Ala
245 250 255
Glu Thr Glu Lys Met Ile Glu Ser Thr Pro Ile Ala Glu Lys Ile Phe
260 265 270
Asp Val Tyr Tyr Phe Ser Ser Cys Leu Ser Gln Lys Gln Ile Glu Glu
275 280 285
Tyr Asn Arg Ile Ile Gly His Tyr Asn Leu Leu Ile Asn Leu Tyr Asn
290 295 300
Gln Ala Lys Arg Ser Glu Gly Lys His Leu Ser Ala Asn Glu Lys Lys
305 310 315 320
Tyr Lys Asp Leu Pro Lys Phe Lys Thr Leu Tyr Lys Gln Ile Gly Cys
325 330 335
Gly Lys Lys Lys Asp Leu Phe Tyr Thr Ile Lys Cys Asp Thr Glu Glu
340 345 350
Glu Ala Asn Lys Ser Arg Asn Glu Gly Lys Glu Ser His Ser Val Glu
355 360 365
Glu Ile Ile Asn Lys Ala Gln Glu Ala Ile Asn Lys Tyr Phe Lys Ser
370 375 380
Asn Asn Asp Cys Glu Asn Ile Asn Thr Val Pro Asp Phe Ile Asn Tyr
385 390 395 400
Ile Leu Thr Lys Glu Asn Tyr Glu Gly Val Tyr Trp Ser Lys Ala Ala
405 410 415
Met Asn Thr Ile Ser Asp Lys Tyr Phe Ala Asn Tyr His Asp Leu Gln
420 425 430
Asp Arg Leu Lys Glu Ala Lys Val Phe Gln Lys Ala Asp Lys Lys Ser
435 440 445
Glu Asp Asp Ile Lys Ile Pro Glu Ala Ile Glu Leu Ser Gly Leu Phe
450 455 460
Gly Val Leu Asp Ser Leu Ala Asp Trp Gln Thr Thr Leu Phe Lys Ser
465 470 475 480
Ser Ile Leu Ser Asn Glu Asp Lys Leu Lys Ile Ile Thr Asp Ser Gln
485 490 495
Thr Pro Ser Glu Ala Leu Leu Lys Met Ile Phe Asn Asp Ile Glu Lys
500 505 510
Asn Met Glu Ser Phe Leu Lys Glu Thr Asn Asp Ile Ile Thr Leu Lys
515 520 525
Lys Tyr Lys Gly Asn Lys Glu Gly Thr Glu Lys Ile Lys Gln Trp Phe
530 535 540
Asp Tyr Thr Leu Ala Ile Asn Arg Met Leu Lys Tyr Phe Leu Val Lys
545 550 555 560
Glu Asn Lys Ile Lys Gly Asn Ser Leu Asp Thr Asn Ile Ser Glu Ala
565 570 575
Leu Lys Thr Leu Ile Tyr Ser Asp Asp Ala Glu Trp Phe Lys Trp Tyr
580 585 590
Asp Ala Leu Arg Asn Tyr Leu Thr Gln Lys Pro Gln Asp Glu Ala Lys
595 600 605
Glu Asn Lys Leu Lys Leu Asn Phe Asp Asn Pro Ser Leu Ala Gly Gly
610 615 620
Trp Asp Val Asn Lys Glu Cys Ser Asn Phe Cys Val Ile Leu Lys Asp
625 630 635 640
Lys Asn Glu Lys Lys Tyr Leu Ala Ile Met Lys Lys Gly Glu Asn Thr
645 650 655
Leu Phe Gln Lys Glu Trp Thr Glu Gly Arg Gly Lys Asn Leu Thr Lys
660 665 670
Lys Ser Asn Pro Leu Phe Glu Ile Asn Asn Cys Glu Ile Leu Ser Lys
675 680 685
Met Glu Tyr Asp Phe Trp Ala Asp Val Ser Lys Met Ile Pro Lys Cys
690 695 700
Ser Thr Gln Leu Lys Ala Val Val Asn His Phe Lys Gln Ser Asp Asn
705 710 715 720
Glu Phe Ile Phe Pro Ile Gly Tyr Lys Val Thr Ser Gly Glu Lys Phe
725 730 735
Arg Glu Glu Cys Lys Ile Ser Lys Gln Asp Phe Glu Leu Asn Asn Lys
740 745 750
Val Phe Asn Lys Asn Glu Leu Ser Val Thr Ala Met Arg Tyr Asp Leu
755 760 765
Ser Ser Thr Gln Glu Lys Gln Tyr Ile Lys Ala Phe Gln Lys Glu Tyr
770 775 780
Trp Glu Leu Leu Phe Lys Gln Glu Lys Arg Asp Thr Lys Leu Thr Asn
785 790 795 800
Asn Glu Ile Phe Asn Glu Trp Ile Asn Phe Cys Asn Lys Lys Tyr Ser
805 810 815
Glu Leu Leu Ser Trp Glu Arg Lys Tyr Lys Asp Ala Leu Thr Asn Trp
820 825 830
Ile Asn Phe Cys Lys Tyr Phe Leu Ser Lys Tyr Pro Lys Thr Thr Leu
835 840 845
Phe Asn Tyr Ser Phe Lys Glu Ser Glu Asn Tyr Asn Ser Leu Asp Glu
850 855 860
Phe Tyr Arg Asp Val Asp Ile Cys Ser Tyr Lys Leu Asn Ile Asn Thr
865 870 875 880
Thr Ile Asn Lys Ser Ile Leu Asp Arg Leu Val Glu Glu Gly Lys Leu
885 890 895
Tyr Leu Phe Glu Ile Lys Asn Gln Asp Ser Asn Asp Gly Lys Ser Ile
900 905 910
Gly His Lys Asn Asn Leu His Thr Ile Tyr Trp Asn Ala Ile Phe Glu
915 920 925
Asn Phe Asp Asn Arg Pro Lys Leu Asn Gly Glu Ala Glu Ile Phe Tyr
930 935 940
Arg Lys Ala Ile Ser Lys Asp Lys Leu Gly Ile Val Lys Gly Lys Lys
945 950 955 960
Thr Lys Asn Gly Thr Glu Ile Ile Lys Asn Tyr Arg Phe Ser Lys Glu
965 970 975
Lys Phe Ile Leu His Val Pro Ile Thr Leu Asn Phe Cys Ser Asn Asn
980 985 990
Glu Tyr Val Asn Asp Ile Val Asn Thr Lys Phe Tyr Asn Phe Ser Asn
995 1000 1005
Leu His Phe Leu Gly Ile Asp Arg Gly Glu Lys His Leu Ala Tyr
1010 1015 1020
Tyr Ser Leu Val Asn Lys Asn Gly Glu Ile Val Asp Gln Gly Thr
1025 1030 1035
Leu Asn Leu Pro Phe Thr Asp Lys Asp Gly Asn Gln Arg Ser Ile
1040 1045 1050
Lys Lys Glu Lys Tyr Phe Tyr Asn Lys Gln Glu Asp Lys Trp Glu
1055 1060 1065
Ala Lys Glu Val Asp Cys Trp Asn Tyr Asn Asp Leu Leu Asp Ala
1070 1075 1080
Met Ala Ser Asn Arg Asp Met Ala Arg Lys Asn Trp Gln Arg Ile
1085 1090 1095
Gly Thr Ile Lys Glu Ala Lys Asn Gly Tyr Val Ser Leu Val Ile
1100 1105 1110
Arg Lys Ile Ala Asp Leu Ala Val Asn Asn Glu Arg Pro Ala Phe
1115 1120 1125
Ile Val Leu Glu Asp Leu Asn Thr Gly Phe Lys Arg Ser Arg Gln
1130 1135 1140
Lys Ile Asp Lys Ser Val Tyr Gln Lys Phe Glu Leu Ala Leu Ala
1145 1150 1155
Lys Lys Leu Asn Phe Leu Val Asp Lys Asn Ala Lys Arg Asp Glu
1160 1165 1170
Ile Gly Ser Pro Thr Lys Ala Leu Gln Leu Thr Pro Pro Val Asn
1175 1180 1185
Asn Tyr Gly Asp Ile Glu Asn Lys Lys Gln Ala Gly Ile Met Leu
1190 1195 1200
Tyr Thr Arg Ala Asn Tyr Thr Ser Gln Thr Asp Pro Ala Thr Gly
1205 1210 1215
Trp Arg Lys Thr Ile Tyr Leu Lys Ala Gly Pro Glu Glu Thr Thr
1220 1225 1230
Tyr Lys Lys Asp Gly Lys Ile Lys Asn Lys Ser Val Lys Asp Gln
1235 1240 1245
Ile Ile Glu Thr Phe Thr Asp Ile Gly Phe Asp Gly Lys Asp Tyr
1250 1255 1260
Tyr Phe Glu Tyr Asp Lys Gly Glu Phe Val Asp Glu Lys Thr Gly
1265 1270 1275
Glu Ile Lys Pro Lys Lys Trp Arg Leu Tyr Ser Gly Glu Asn Gly
1280 1285 1290
Lys Ser Leu Asp Arg Phe Arg Gly Glu Arg Glu Lys Asp Lys Tyr
1295 1300 1305
Glu Trp Lys Ile Asp Lys Ile Asp Ile Val Lys Ile Leu Asp Asp
1310 1315 1320
Leu Phe Val Asn Phe Asp Lys Asn Ile Ser Leu Leu Lys Gln Leu
1325 1330 1335
Lys Glu Gly Val Glu Leu Thr Arg Asn Asn Glu His Gly Thr Gly
1340 1345 1350
Glu Ser Leu Arg Phe Ala Ile Asn Leu Ile Gln Gln Ile Arg Asn
1355 1360 1365
Thr Gly Asn Asn Glu Arg Asp Asn Asp Phe Ile Leu Ser Pro Val
1370 1375 1380
Arg Asp Glu Asn Gly Lys His Phe Asp Ser Arg Glu Tyr Trp Asp
1385 1390 1395
Lys Glu Thr Lys Gly Glu Lys Ile Ser Met Pro Ser Ser Gly Asp
1400 1405 1410
Ala Asn Gly Ala Phe Asn Ile Ala Arg Lys Gly Ile Ile Met Asn
1415 1420 1425
Ala His Ile Leu Ala Asn Ser Asp Ser Lys Asp Leu Ser Leu Phe
1430 1435 1440
Val Ser Asp Glu Glu Trp Asp Leu His Leu Asn Asn Lys Thr Glu
1445 1450 1455
Trp Lys Lys Gln Leu Asn Ile Phe Ser Ser Arg Lys Ala Met Ala
1460 1465 1470
Lys Arg Lys Lys Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln
1475 1480 1485
Ala Lys Lys Lys Lys Gly Ser Tyr Pro Tyr Asp Val Pro Asp Tyr
1490 1495 1500
Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Tyr Pro Tyr Asp Val
1505 1510 1515
Pro Asp Tyr Ala
1520
<210> 11
<211> 1397
<212> PRT
<213> 俭菌总门细菌GWC2011_GWC2_44_17
<220>
<221> MISC_FEATURE
<222> (1)..(1397)
<223> PbCpf1; pY008
<400> 11
Met Glu Asn Ile Phe Asp Gln Phe Ile Gly Lys Tyr Ser Leu Ser Lys
1 5 10 15
Thr Leu Arg Phe Glu Leu Lys Pro Val Gly Lys Thr Glu Asp Phe Leu
20 25 30
Lys Ile Asn Lys Val Phe Glu Lys Asp Gln Thr Ile Asp Asp Ser Tyr
35 40 45
Asn Gln Ala Lys Phe Tyr Phe Asp Ser Leu His Gln Lys Phe Ile Asp
50 55 60
Ala Ala Leu Ala Ser Asp Lys Thr Ser Glu Leu Ser Phe Gln Asn Phe
65 70 75 80
Ala Asp Val Leu Glu Lys Gln Asn Lys Ile Ile Leu Asp Lys Lys Arg
85 90 95
Glu Met Gly Ala Leu Arg Lys Arg Asp Lys Asn Ala Val Gly Ile Asp
100 105 110
Arg Leu Gln Lys Glu Ile Asn Asp Ala Glu Asp Ile Ile Gln Lys Glu
115 120 125
Lys Glu Lys Ile Tyr Lys Asp Val Arg Thr Leu Phe Asp Asn Glu Ala
130 135 140
Glu Ser Trp Lys Thr Tyr Tyr Gln Glu Arg Glu Val Asp Gly Lys Lys
145 150 155 160
Ile Thr Phe Ser Lys Ala Asp Leu Lys Gln Lys Gly Ala Asp Phe Leu
165 170 175
Thr Ala Ala Gly Ile Leu Lys Val Leu Lys Tyr Glu Phe Pro Glu Glu
180 185 190
Lys Glu Lys Glu Phe Gln Ala Lys Asn Gln Pro Ser Leu Phe Val Glu
195 200 205
Glu Lys Glu Asn Pro Gly Gln Lys Arg Tyr Ile Phe Asp Ser Phe Asp
210 215 220
Lys Phe Ala Gly Tyr Leu Thr Lys Phe Gln Gln Thr Lys Lys Asn Leu
225 230 235 240
Tyr Ala Ala Asp Gly Thr Ser Thr Ala Val Ala Thr Arg Ile Ala Asp
245 250 255
Asn Phe Ile Ile Phe His Gln Asn Thr Lys Val Phe Arg Asp Lys Tyr
260 265 270
Lys Asn Asn His Thr Asp Leu Gly Phe Asp Glu Glu Asn Ile Phe Glu
275 280 285
Ile Glu Arg Tyr Lys Asn Cys Leu Leu Gln Arg Glu Ile Glu His Ile
290 295 300
Lys Asn Glu Asn Ser Tyr Asn Lys Ile Ile Gly Arg Ile Asn Lys Lys
305 310 315 320
Ile Lys Glu Tyr Arg Asp Gln Lys Ala Lys Asp Thr Lys Leu Thr Lys
325 330 335
Ser Asp Phe Pro Phe Phe Lys Asn Leu Asp Lys Gln Ile Leu Gly Glu
340 345 350
Val Glu Lys Glu Lys Gln Leu Ile Glu Lys Thr Arg Glu Lys Thr Glu
355 360 365
Glu Asp Val Leu Ile Glu Arg Phe Lys Glu Phe Ile Glu Asn Asn Glu
370 375 380
Glu Arg Phe Thr Ala Ala Lys Lys Leu Met Asn Ala Phe Cys Asn Gly
385 390 395 400
Glu Phe Glu Ser Glu Tyr Glu Gly Ile Tyr Leu Lys Asn Lys Ala Ile
405 410 415
Asn Thr Ile Ser Arg Arg Trp Phe Val Ser Asp Arg Asp Phe Glu Leu
420 425 430
Lys Leu Pro Gln Gln Lys Ser Lys Asn Lys Ser Glu Lys Asn Glu Pro
435 440 445
Lys Val Lys Lys Phe Ile Ser Ile Ala Glu Ile Lys Asn Ala Val Glu
450 455 460
Glu Leu Asp Gly Asp Ile Phe Lys Ala Val Phe Tyr Asp Lys Lys Ile
465 470 475 480
Ile Ala Gln Gly Gly Ser Lys Leu Glu Gln Phe Leu Val Ile Trp Lys
485 490 495
Tyr Glu Phe Glu Tyr Leu Phe Arg Asp Ile Glu Arg Glu Asn Gly Glu
500 505 510
Lys Leu Leu Gly Tyr Asp Ser Cys Leu Lys Ile Ala Lys Gln Leu Gly
515 520 525
Ile Phe Pro Gln Glu Lys Glu Ala Arg Glu Lys Ala Thr Ala Val Ile
530 535 540
Lys Asn Tyr Ala Asp Ala Gly Leu Gly Ile Phe Gln Met Met Lys Tyr
545 550 555 560
Phe Ser Leu Asp Asp Lys Asp Arg Lys Asn Thr Pro Gly Gln Leu Ser
565 570 575
Thr Asn Phe Tyr Ala Glu Tyr Asp Gly Tyr Tyr Lys Asp Phe Glu Phe
580 585 590
Ile Lys Tyr Tyr Asn Glu Phe Arg Asn Phe Ile Thr Lys Lys Pro Phe
595 600 605
Asp Glu Asp Lys Ile Lys Leu Asn Phe Glu Asn Gly Ala Leu Leu Lys
610 615 620
Gly Trp Asp Glu Asn Lys Glu Tyr Asp Phe Met Gly Val Ile Leu Lys
625 630 635 640
Lys Glu Gly Arg Leu Tyr Leu Gly Ile Met His Lys Asn His Arg Lys
645 650 655
Leu Phe Gln Ser Met Gly Asn Ala Lys Gly Asp Asn Ala Asn Arg Tyr
660 665 670
Gln Lys Met Ile Tyr Lys Gln Ile Ala Asp Ala Ser Lys Asp Val Pro
675 680 685
Arg Leu Leu Leu Thr Ser Lys Lys Ala Met Glu Lys Phe Lys Pro Ser
690 695 700
Gln Glu Ile Leu Arg Ile Lys Lys Glu Lys Thr Phe Lys Arg Glu Ser
705 710 715 720
Lys Asn Phe Ser Leu Arg Asp Leu His Ala Leu Ile Glu Tyr Tyr Arg
725 730 735
Asn Cys Ile Pro Gln Tyr Ser Asn Trp Ser Phe Tyr Asp Phe Gln Phe
740 745 750
Gln Asp Thr Gly Lys Tyr Gln Asn Ile Lys Glu Phe Thr Asp Asp Val
755 760 765
Gln Lys Tyr Gly Tyr Lys Ile Ser Phe Arg Asp Ile Asp Asp Glu Tyr
770 775 780
Ile Asn Gln Ala Leu Asn Glu Gly Lys Met Tyr Leu Phe Glu Val Val
785 790 795 800
Asn Lys Asp Ile Tyr Asn Thr Lys Asn Gly Ser Lys Asn Leu His Thr
805 810 815
Leu Tyr Phe Glu His Ile Leu Ser Ala Glu Asn Leu Asn Asp Pro Val
820 825 830
Phe Lys Leu Ser Gly Met Ala Glu Ile Phe Gln Arg Gln Pro Ser Val
835 840 845
Asn Glu Arg Glu Lys Ile Thr Thr Gln Lys Asn Gln Cys Ile Leu Asp
850 855 860
Lys Gly Asp Arg Ala Tyr Lys Tyr Arg Arg Tyr Thr Glu Lys Lys Ile
865 870 875 880
Met Phe His Met Ser Leu Val Leu Asn Thr Gly Lys Gly Glu Ile Lys
885 890 895
Gln Val Gln Phe Asn Lys Ile Ile Asn Gln Arg Ile Ser Ser Ser Asp
900 905 910
Asn Glu Met Arg Val Asn Val Ile Gly Ile Asp Arg Gly Glu Lys Asn
915 920 925
Leu Leu Tyr Tyr Ser Val Val Lys Gln Asn Gly Glu Ile Ile Glu Gln
930 935 940
Ala Ser Leu Asn Glu Ile Asn Gly Val Asn Tyr Arg Asp Lys Leu Ile
945 950 955 960
Glu Arg Glu Lys Glu Arg Leu Lys Asn Arg Gln Ser Trp Lys Pro Val
965 970 975
Val Lys Ile Lys Asp Leu Lys Lys Gly Tyr Ile Ser His Val Ile His
980 985 990
Lys Ile Cys Gln Leu Ile Glu Lys Tyr Ser Ala Ile Val Val Leu Glu
995 1000 1005
Asp Leu Asn Met Arg Phe Lys Gln Ile Arg Gly Gly Ile Glu Arg
1010 1015 1020
Ser Val Tyr Gln Gln Phe Glu Lys Ala Leu Ile Asp Lys Leu Gly
1025 1030 1035
Tyr Leu Val Phe Lys Asp Asn Arg Asp Leu Arg Ala Pro Gly Gly
1040 1045 1050
Val Leu Asn Gly Tyr Gln Leu Ser Ala Pro Phe Val Ser Phe Glu
1055 1060 1065
Lys Met Arg Lys Gln Thr Gly Ile Leu Phe Tyr Thr Gln Ala Glu
1070 1075 1080
Tyr Thr Ser Lys Thr Asp Pro Ile Thr Gly Phe Arg Lys Asn Val
1085 1090 1095
Tyr Ile Ser Asn Ser Ala Ser Leu Asp Lys Ile Lys Glu Ala Val
1100 1105 1110
Lys Lys Phe Asp Ala Ile Gly Trp Asp Gly Lys Glu Gln Ser Tyr
1115 1120 1125
Phe Phe Lys Tyr Asn Pro Tyr Asn Leu Ala Asp Glu Lys Tyr Lys
1130 1135 1140
Asn Ser Thr Val Ser Lys Glu Trp Ala Ile Phe Ala Ser Ala Pro
1145 1150 1155
Arg Ile Arg Arg Gln Lys Gly Glu Asp Gly Tyr Trp Lys Tyr Asp
1160 1165 1170
Arg Val Lys Val Asn Glu Glu Phe Glu Lys Leu Leu Lys Val Trp
1175 1180 1185
Asn Phe Val Asn Pro Lys Ala Thr Asp Ile Lys Gln Glu Ile Ile
1190 1195 1200
Lys Lys Glu Lys Ala Gly Asp Leu Gln Gly Glu Lys Glu Leu Asp
1205 1210 1215
Gly Arg Leu Arg Asn Phe Trp His Ser Phe Ile Tyr Leu Phe Asn
1220 1225 1230
Leu Val Leu Glu Leu Arg Asn Ser Phe Ser Leu Gln Ile Lys Ile
1235 1240 1245
Lys Ala Gly Glu Val Ile Ala Val Asp Glu Gly Val Asp Phe Ile
1250 1255 1260
Ala Ser Pro Val Lys Pro Phe Phe Thr Thr Pro Asn Pro Tyr Ile
1265 1270 1275
Pro Ser Asn Leu Cys Trp Leu Ala Val Glu Asn Ala Asp Ala Asn
1280 1285 1290
Gly Ala Tyr Asn Ile Ala Arg Lys Gly Val Met Ile Leu Lys Lys
1295 1300 1305
Ile Arg Glu His Ala Lys Lys Asp Pro Glu Phe Lys Lys Leu Pro
1310 1315 1320
Asn Leu Phe Ile Ser Asn Ala Glu Trp Asp Glu Ala Ala Arg Asp
1325 1330 1335
Trp Gly Lys Tyr Ala Gly Thr Thr Ala Leu Asn Leu Asp His Lys
1340 1345 1350
Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys
1355 1360 1365
Gly Ser Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Tyr Pro Tyr Asp
1370 1375 1380
Val Pro Asp Tyr Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
1385 1390 1395
<210> 12
<211> 1295
<212> PRT
<213> 史密斯氏菌SC_K08D17
<220>
<221> MISC_FEATURE
<222> (1)..(1295)
<223> SsCpf1; pY009
<400> 12
Met Gln Thr Leu Phe Glu Asn Phe Thr Asn Gln Tyr Pro Val Ser Lys
1 5 10 15
Thr Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Lys Asp Phe Ile
20 25 30
Glu Gln Lys Gly Leu Leu Lys Lys Asp Glu Asp Arg Ala Glu Lys Tyr
35 40 45
Lys Lys Val Lys Asn Ile Ile Asp Glu Tyr His Lys Asp Phe Ile Glu
50 55 60
Lys Ser Leu Asn Gly Leu Lys Leu Asp Gly Leu Glu Lys Tyr Lys Thr
65 70 75 80
Leu Tyr Leu Lys Gln Glu Lys Asp Asp Lys Asp Lys Lys Ala Phe Asp
85 90 95
Lys Glu Lys Glu Asn Leu Arg Lys Gln Ile Ala Asn Ala Phe Arg Asn
100 105 110
Asn Glu Lys Phe Lys Thr Leu Phe Ala Lys Glu Leu Ile Lys Asn Asp
115 120 125
Leu Met Ser Phe Ala Cys Glu Glu Asp Lys Lys Asn Val Lys Glu Phe
130 135 140
Glu Ala Phe Thr Thr Tyr Phe Thr Gly Phe His Gln Asn Arg Ala Asn
145 150 155 160
Met Tyr Val Ala Asp Glu Lys Arg Thr Ala Ile Ala Ser Arg Leu Ile
165 170 175
His Glu Asn Leu Pro Lys Phe Ile Asp Asn Ile Lys Ile Phe Glu Lys
180 185 190
Met Lys Lys Glu Ala Pro Glu Leu Leu Ser Pro Phe Asn Gln Thr Leu
195 200 205
Lys Asp Met Lys Asp Val Ile Lys Gly Thr Thr Leu Glu Glu Ile Phe
210 215 220
Ser Leu Asp Tyr Phe Asn Lys Thr Leu Thr Gln Ser Gly Ile Asp Ile
225 230 235 240
Tyr Asn Ser Val Ile Gly Gly Arg Thr Pro Glu Glu Gly Lys Thr Lys
245 250 255
Ile Lys Gly Leu Asn Glu Tyr Ile Asn Thr Asp Phe Asn Gln Lys Gln
260 265 270
Thr Asp Lys Lys Lys Arg Gln Pro Lys Phe Lys Gln Leu Tyr Lys Gln
275 280 285
Ile Leu Ser Asp Arg Gln Ser Leu Ser Phe Ile Ala Glu Ala Phe Lys
290 295 300
Asn Asp Thr Glu Ile Leu Glu Ala Ile Glu Lys Phe Tyr Val Asn Glu
305 310 315 320
Leu Leu His Phe Ser Asn Glu Gly Lys Ser Thr Asn Val Leu Asp Ala
325 330 335
Ile Lys Asn Ala Val Ser Asn Leu Glu Ser Phe Asn Leu Thr Lys Met
340 345 350
Tyr Phe Arg Ser Gly Ala Ser Leu Thr Asp Val Ser Arg Lys Val Phe
355 360 365
Gly Glu Trp Ser Ile Ile Asn Arg Ala Leu Asp Asn Tyr Tyr Ala Thr
370 375 380
Thr Tyr Pro Ile Lys Pro Arg Glu Lys Ser Glu Lys Tyr Glu Glu Arg
385 390 395 400
Lys Glu Lys Trp Leu Lys Gln Asp Phe Asn Val Ser Leu Ile Gln Thr
405 410 415
Ala Ile Asp Glu Tyr Asp Asn Glu Thr Val Lys Gly Lys Asn Ser Gly
420 425 430
Lys Val Ile Ala Asp Tyr Phe Ala Lys Phe Cys Asp Asp Lys Glu Thr
435 440 445
Asp Leu Ile Gln Lys Val Asn Glu Gly Tyr Ile Ala Val Lys Asp Leu
450 455 460
Leu Asn Thr Pro Cys Pro Glu Asn Glu Lys Leu Gly Ser Asn Lys Asp
465 470 475 480
Gln Val Lys Gln Ile Lys Ala Phe Met Asp Ser Ile Met Asp Ile Met
485 490 495
His Phe Val Arg Pro Leu Ser Leu Lys Asp Thr Asp Lys Glu Lys Asp
500 505 510
Glu Thr Phe Tyr Ser Leu Phe Thr Pro Leu Tyr Asp His Leu Thr Gln
515 520 525
Thr Ile Ala Leu Tyr Asn Lys Val Arg Asn Tyr Leu Thr Gln Lys Pro
530 535 540
Tyr Ser Thr Glu Lys Ile Lys Leu Asn Phe Glu Asn Ser Thr Leu Leu
545 550 555 560
Gly Gly Trp Asp Leu Asn Lys Glu Thr Asp Asn Thr Ala Ile Ile Leu
565 570 575
Arg Lys Asp Asn Leu Tyr Tyr Leu Gly Ile Met Asp Lys Arg His Asn
580 585 590
Arg Ile Phe Arg Asn Val Pro Lys Ala Asp Lys Lys Asp Phe Cys Tyr
595 600 605
Glu Lys Met Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro
610 615 620
Lys Val Phe Phe Ser Gln Ser Arg Ile Gln Glu Phe Thr Pro Ser Ala
625 630 635 640
Lys Leu Leu Glu Asn Tyr Ala Asn Glu Thr His Lys Lys Gly Asp Asn
645 650 655
Phe Asn Leu Asn His Cys His Lys Leu Ile Asp Phe Phe Lys Asp Ser
660 665 670
Ile Asn Lys His Glu Asp Trp Lys Asn Phe Asp Phe Arg Phe Ser Ala
675 680 685
Thr Ser Thr Tyr Ala Asp Leu Ser Gly Phe Tyr His Glu Val Glu His
690 695 700
Gln Gly Tyr Lys Ile Ser Phe Gln Ser Val Ala Asp Ser Phe Ile Asp
705 710 715 720
Asp Leu Val Asn Glu Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys
725 730 735
Asp Phe Ser Pro Phe Ser Lys Gly Lys Pro Asn Leu His Thr Leu Tyr
740 745 750
Trp Lys Met Leu Phe Asp Glu Asn Asn Leu Lys Asp Val Val Tyr Lys
755 760 765
Leu Asn Gly Glu Ala Glu Val Phe Tyr Arg Lys Lys Ser Ile Ala Glu
770 775 780
Lys Asn Thr Thr Ile His Lys Ala Asn Glu Ser Ile Ile Asn Lys Asn
785 790 795 800
Pro Asp Asn Pro Lys Ala Thr Ser Thr Phe Asn Tyr Asp Ile Val Lys
805 810 815
Asp Lys Arg Tyr Thr Ile Asp Lys Phe Gln Phe His Ile Pro Ile Thr
820 825 830
Met Asn Phe Lys Ala Glu Gly Ile Phe Asn Met Asn Gln Arg Val Asn
835 840 845
Gln Phe Leu Lys Ala Asn Pro Asp Ile Asn Ile Ile Gly Ile Asp Arg
850 855 860
Gly Glu Arg His Leu Leu Tyr Tyr Ala Leu Ile Asn Gln Lys Gly Lys
865 870 875 880
Ile Leu Lys Gln Asp Thr Leu Asn Val Ile Ala Asn Glu Lys Gln Lys
885 890 895
Val Asp Tyr His Asn Leu Leu Asp Lys Lys Glu Gly Asp Arg Ala Thr
900 905 910
Ala Arg Gln Glu Trp Gly Val Ile Glu Thr Ile Lys Glu Leu Lys Glu
915 920 925
Gly Tyr Leu Ser Gln Val Ile His Lys Leu Thr Asp Leu Met Ile Glu
930 935 940
Asn Asn Ala Ile Ile Val Met Glu Asp Leu Asn Phe Gly Phe Lys Arg
945 950 955 960
Gly Arg Gln Lys Val Glu Lys Gln Val Tyr Gln Lys Phe Glu Lys Met
965 970 975
Leu Ile Asp Lys Leu Asn Tyr Leu Val Asp Lys Asn Lys Lys Ala Asn
980 985 990
Glu Leu Gly Gly Leu Leu Asn Ala Phe Gln Leu Ala Asn Lys Phe Glu
995 1000 1005
Ser Phe Gln Lys Met Gly Lys Gln Asn Gly Phe Ile Phe Tyr Val
1010 1015 1020
Pro Ala Trp Asn Thr Ser Lys Thr Asp Pro Ala Thr Gly Phe Ile
1025 1030 1035
Asp Phe Leu Lys Pro Arg Tyr Glu Asn Leu Asn Gln Ala Lys Asp
1040 1045 1050
Phe Phe Glu Lys Phe Asp Ser Ile Arg Leu Asn Ser Lys Ala Asp
1055 1060 1065
Tyr Phe Glu Phe Ala Phe Asp Phe Lys Asn Phe Thr Glu Lys Ala
1070 1075 1080
Asp Gly Gly Arg Thr Lys Trp Thr Val Cys Thr Thr Asn Glu Asp
1085 1090 1095
Arg Tyr Ala Trp Asn Arg Ala Leu Asn Asn Asn Arg Gly Ser Gln
1100 1105 1110
Glu Lys Tyr Asp Ile Thr Ala Glu Leu Lys Ser Leu Phe Asp Gly
1115 1120 1125
Lys Val Asp Tyr Lys Ser Gly Lys Asp Leu Lys Gln Gln Ile Ala
1130 1135 1140
Ser Gln Glu Ser Ala Asp Phe Phe Lys Ala Leu Met Lys Asn Leu
1145 1150 1155
Ser Ile Thr Leu Ser Leu Arg His Asn Asn Gly Glu Lys Gly Asp
1160 1165 1170
Asn Glu Gln Asp Tyr Ile Leu Ser Pro Val Ala Asp Ser Lys Gly
1175 1180 1185
Arg Phe Phe Asp Ser Arg Lys Ala Asp Asp Asp Met Pro Lys Asn
1190 1195 1200
Ala Asp Ala Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Leu Trp
1205 1210 1215
Cys Leu Glu Gln Ile Ser Lys Thr Asp Asp Leu Lys Lys Val Lys
1220 1225 1230
Leu Ala Ile Ser Asn Lys Glu Trp Leu Glu Phe Val Gln Thr Leu
1235 1240 1245
Lys Gly Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys
1250 1255 1260
Lys Lys Lys Gly Ser Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Tyr
1265 1270 1275
Pro Tyr Asp Val Pro Asp Tyr Ala Tyr Pro Tyr Asp Val Pro Asp
1280 1285 1290
Tyr Ala
1295
<210> 13
<211> 1352
<212> PRT
<213> 氨基酸球菌BV3L6
<220>
<221> MISC_FEATURE
<222> (1)..(1352)
<223> AsCpf1; pY010
<400> 13
Met Thr Gln Phe Glu Gly Phe Thr Asn Leu Tyr Gln Val Ser Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Lys His Ile Gln
20 25 30
Glu Gln Gly Phe Ile Glu Glu Asp Lys Ala Arg Asn Asp His Tyr Lys
35 40 45
Glu Leu Lys Pro Ile Ile Asp Arg Ile Tyr Lys Thr Tyr Ala Asp Gln
50 55 60
Cys Leu Gln Leu Val Gln Leu Asp Trp Glu Asn Leu Ser Ala Ala Ile
65 70 75 80
Asp Ser Tyr Arg Lys Glu Lys Thr Glu Glu Thr Arg Asn Ala Leu Ile
85 90 95
Glu Glu Gln Ala Thr Tyr Arg Asn Ala Ile His Asp Tyr Phe Ile Gly
100 105 110
Arg Thr Asp Asn Leu Thr Asp Ala Ile Asn Lys Arg His Ala Glu Ile
115 120 125
Tyr Lys Gly Leu Phe Lys Ala Glu Leu Phe Asn Gly Lys Val Leu Lys
130 135 140
Gln Leu Gly Thr Val Thr Thr Thr Glu His Glu Asn Ala Leu Leu Arg
145 150 155 160
Ser Phe Asp Lys Phe Thr Thr Tyr Phe Ser Gly Phe Tyr Glu Asn Arg
165 170 175
Lys Asn Val Phe Ser Ala Glu Asp Ile Ser Thr Ala Ile Pro His Arg
180 185 190
Ile Val Gln Asp Asn Phe Pro Lys Phe Lys Glu Asn Cys His Ile Phe
195 200 205
Thr Arg Leu Ile Thr Ala Val Pro Ser Leu Arg Glu His Phe Glu Asn
210 215 220
Val Lys Lys Ala Ile Gly Ile Phe Val Ser Thr Ser Ile Glu Glu Val
225 230 235 240
Phe Ser Phe Pro Phe Tyr Asn Gln Leu Leu Thr Gln Thr Gln Ile Asp
245 250 255
Leu Tyr Asn Gln Leu Leu Gly Gly Ile Ser Arg Glu Ala Gly Thr Glu
260 265 270
Lys Ile Lys Gly Leu Asn Glu Val Leu Asn Leu Ala Ile Gln Lys Asn
275 280 285
Asp Glu Thr Ala His Ile Ile Ala Ser Leu Pro His Arg Phe Ile Pro
290 295 300
Leu Phe Lys Gln Ile Leu Ser Asp Arg Asn Thr Leu Ser Phe Ile Leu
305 310 315 320
Glu Glu Phe Lys Ser Asp Glu Glu Val Ile Gln Ser Phe Cys Lys Tyr
325 330 335
Lys Thr Leu Leu Arg Asn Glu Asn Val Leu Glu Thr Ala Glu Ala Leu
340 345 350
Phe Asn Glu Leu Asn Ser Ile Asp Leu Thr His Ile Phe Ile Ser His
355 360 365
Lys Lys Leu Glu Thr Ile Ser Ser Ala Leu Cys Asp His Trp Asp Thr
370 375 380
Leu Arg Asn Ala Leu Tyr Glu Arg Arg Ile Ser Glu Leu Thr Gly Lys
385 390 395 400
Ile Thr Lys Ser Ala Lys Glu Lys Val Gln Arg Ser Leu Lys His Glu
405 410 415
Asp Ile Asn Leu Gln Glu Ile Ile Ser Ala Ala Gly Lys Glu Leu Ser
420 425 430
Glu Ala Phe Lys Gln Lys Thr Ser Glu Ile Leu Ser His Ala His Ala
435 440 445
Ala Leu Asp Gln Pro Leu Pro Thr Thr Leu Lys Lys Gln Glu Glu Lys
450 455 460
Glu Ile Leu Lys Ser Gln Leu Asp Ser Leu Leu Gly Leu Tyr His Leu
465 470 475 480
Leu Asp Trp Phe Ala Val Asp Glu Ser Asn Glu Val Asp Pro Glu Phe
485 490 495
Ser Ala Arg Leu Thr Gly Ile Lys Leu Glu Met Glu Pro Ser Leu Ser
500 505 510
Phe Tyr Asn Lys Ala Arg Asn Tyr Ala Thr Lys Lys Pro Tyr Ser Val
515 520 525
Glu Lys Phe Lys Leu Asn Phe Gln Met Pro Thr Leu Ala Ser Gly Trp
530 535 540
Asp Val Asn Lys Glu Lys Asn Asn Gly Ala Ile Leu Phe Val Lys Asn
545 550 555 560
Gly Leu Tyr Tyr Leu Gly Ile Met Pro Lys Gln Lys Gly Arg Tyr Lys
565 570 575
Ala Leu Ser Phe Glu Pro Thr Glu Lys Thr Ser Glu Gly Phe Asp Lys
580 585 590
Met Tyr Tyr Asp Tyr Phe Pro Asp Ala Ala Lys Met Ile Pro Lys Cys
595 600 605
Ser Thr Gln Leu Lys Ala Val Thr Ala His Phe Gln Thr His Thr Thr
610 615 620
Pro Ile Leu Leu Ser Asn Asn Phe Ile Glu Pro Leu Glu Ile Thr Lys
625 630 635 640
Glu Ile Tyr Asp Leu Asn Asn Pro Glu Lys Glu Pro Lys Lys Phe Gln
645 650 655
Thr Ala Tyr Ala Lys Lys Thr Gly Asp Gln Lys Gly Tyr Arg Glu Ala
660 665 670
Leu Cys Lys Trp Ile Asp Phe Thr Arg Asp Phe Leu Ser Lys Tyr Thr
675 680 685
Lys Thr Thr Ser Ile Asp Leu Ser Ser Leu Arg Pro Ser Ser Gln Tyr
690 695 700
Lys Asp Leu Gly Glu Tyr Tyr Ala Glu Leu Asn Pro Leu Leu Tyr His
705 710 715 720
Ile Ser Phe Gln Arg Ile Ala Glu Lys Glu Ile Met Asp Ala Val Glu
725 730 735
Thr Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ala Lys
740 745 750
Gly His His Gly Lys Pro Asn Leu His Thr Leu Tyr Trp Thr Gly Leu
755 760 765
Phe Ser Pro Glu Asn Leu Ala Lys Thr Ser Ile Lys Leu Asn Gly Gln
770 775 780
Ala Glu Leu Phe Tyr Arg Pro Lys Ser Arg Met Lys Arg Met Ala His
785 790 795 800
Arg Leu Gly Glu Lys Met Leu Asn Lys Lys Leu Lys Asp Gln Lys Thr
805 810 815
Pro Ile Pro Asp Thr Leu Tyr Gln Glu Leu Tyr Asp Tyr Val Asn His
820 825 830
Arg Leu Ser His Asp Leu Ser Asp Glu Ala Arg Ala Leu Leu Pro Asn
835 840 845
Val Ile Thr Lys Glu Val Ser His Glu Ile Ile Lys Asp Arg Arg Phe
850 855 860
Thr Ser Asp Lys Phe Phe Phe His Val Pro Ile Thr Leu Asn Tyr Gln
865 870 875 880
Ala Ala Asn Ser Pro Ser Lys Phe Asn Gln Arg Val Asn Ala Tyr Leu
885 890 895
Lys Glu His Pro Glu Thr Pro Ile Ile Gly Ile Asp Arg Gly Glu Arg
900 905 910
Asn Leu Ile Tyr Ile Thr Val Ile Asp Ser Thr Gly Lys Ile Leu Glu
915 920 925
Gln Arg Ser Leu Asn Thr Ile Gln Gln Phe Asp Tyr Gln Lys Lys Leu
930 935 940
Asp Asn Arg Glu Lys Glu Arg Val Ala Ala Arg Gln Ala Trp Ser Val
945 950 955 960
Val Gly Thr Ile Lys Asp Leu Lys Gln Gly Tyr Leu Ser Gln Val Ile
965 970 975
His Glu Ile Val Asp Leu Met Ile His Tyr Gln Ala Val Val Val Leu
980 985 990
Glu Asn Leu Asn Phe Gly Phe Lys Ser Lys Arg Thr Gly Ile Ala Glu
995 1000 1005
Lys Ala Val Tyr Gln Gln Phe Glu Lys Met Leu Ile Asp Lys Leu
1010 1015 1020
Asn Cys Leu Val Leu Lys Asp Tyr Pro Ala Glu Lys Val Gly Gly
1025 1030 1035
Val Leu Asn Pro Tyr Gln Leu Thr Asp Gln Phe Thr Ser Phe Ala
1040 1045 1050
Lys Met Gly Thr Gln Ser Gly Phe Leu Phe Tyr Val Pro Ala Pro
1055 1060 1065
Tyr Thr Ser Lys Ile Asp Pro Leu Thr Gly Phe Val Asp Pro Phe
1070 1075 1080
Val Trp Lys Thr Ile Lys Asn His Glu Ser Arg Lys His Phe Leu
1085 1090 1095
Glu Gly Phe Asp Phe Leu His Tyr Asp Val Lys Thr Gly Asp Phe
1100 1105 1110
Ile Leu His Phe Lys Met Asn Arg Asn Leu Ser Phe Gln Arg Gly
1115 1120 1125
Leu Pro Gly Phe Met Pro Ala Trp Asp Ile Val Phe Glu Lys Asn
1130 1135 1140
Glu Thr Gln Phe Asp Ala Lys Gly Thr Pro Phe Ile Ala Gly Lys
1145 1150 1155
Arg Ile Val Pro Val Ile Glu Asn His Arg Phe Thr Gly Arg Tyr
1160 1165 1170
Arg Asp Leu Tyr Pro Ala Asn Glu Leu Ile Ala Leu Leu Glu Glu
1175 1180 1185
Lys Gly Ile Val Phe Arg Asp Gly Ser Asn Ile Leu Pro Lys Leu
1190 1195 1200
Leu Glu Asn Asp Asp Ser His Ala Ile Asp Thr Met Val Ala Leu
1205 1210 1215
Ile Arg Ser Val Leu Gln Met Arg Asn Ser Asn Ala Ala Thr Gly
1220 1225 1230
Glu Asp Tyr Ile Asn Ser Pro Val Arg Asp Leu Asn Gly Val Cys
1235 1240 1245
Phe Asp Ser Arg Phe Gln Asn Pro Glu Trp Pro Met Asp Ala Asp
1250 1255 1260
Ala Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Gln Leu Leu Leu
1265 1270 1275
Asn His Leu Lys Glu Ser Lys Asp Leu Lys Leu Gln Asn Gly Ile
1280 1285 1290
Ser Asn Gln Asp Trp Leu Ala Tyr Ile Gln Glu Leu Arg Asn Lys
1295 1300 1305
Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys
1310 1315 1320
Gly Ser Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Tyr Pro Tyr Asp
1325 1330 1335
Val Pro Asp Tyr Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
1340 1345 1350
<210> 14
<211> 1251
<212> PRT
<213> 毛螺菌科细菌MA2020
<220>
<221> MISC_FEATURE
<222> (1)..(1251)
<223> Lb2Cpf1; pY011
<400> 14
Met Tyr Tyr Glu Ser Leu Thr Lys Gln Tyr Pro Val Ser Lys Thr Ile
1 5 10 15
Arg Asn Glu Leu Ile Pro Ile Gly Lys Thr Leu Asp Asn Ile Arg Gln
20 25 30
Asn Asn Ile Leu Glu Ser Asp Val Lys Arg Lys Gln Asn Tyr Glu His
35 40 45
Val Lys Gly Ile Leu Asp Glu Tyr His Lys Gln Leu Ile Asn Glu Ala
50 55 60
Leu Asp Asn Cys Thr Leu Pro Ser Leu Lys Ile Ala Ala Glu Ile Tyr
65 70 75 80
Leu Lys Asn Gln Lys Glu Val Ser Asp Arg Glu Asp Phe Asn Lys Thr
85 90 95
Gln Asp Leu Leu Arg Lys Glu Val Val Glu Lys Leu Lys Ala His Glu
100 105 110
Asn Phe Thr Lys Ile Gly Lys Lys Asp Ile Leu Asp Leu Leu Glu Lys
115 120 125
Leu Pro Ser Ile Ser Glu Asp Asp Tyr Asn Ala Leu Glu Ser Phe Arg
130 135 140
Asn Phe Tyr Thr Tyr Phe Thr Ser Tyr Asn Lys Val Arg Glu Asn Leu
145 150 155 160
Tyr Ser Asp Lys Glu Lys Ser Ser Thr Val Ala Tyr Arg Leu Ile Asn
165 170 175
Glu Asn Phe Pro Lys Phe Leu Asp Asn Val Lys Ser Tyr Arg Phe Val
180 185 190
Lys Thr Ala Gly Ile Leu Ala Asp Gly Leu Gly Glu Glu Glu Gln Asp
195 200 205
Ser Leu Phe Ile Val Glu Thr Phe Asn Lys Thr Leu Thr Gln Asp Gly
210 215 220
Ile Asp Thr Tyr Asn Ser Gln Val Gly Lys Ile Asn Ser Ser Ile Asn
225 230 235 240
Leu Tyr Asn Gln Lys Asn Gln Lys Ala Asn Gly Phe Arg Lys Ile Pro
245 250 255
Lys Met Lys Met Leu Tyr Lys Gln Ile Leu Ser Asp Arg Glu Glu Ser
260 265 270
Phe Ile Asp Glu Phe Gln Ser Asp Glu Val Leu Ile Asp Asn Val Glu
275 280 285
Ser Tyr Gly Ser Val Leu Ile Glu Ser Leu Lys Ser Ser Lys Val Ser
290 295 300
Ala Phe Phe Asp Ala Leu Arg Glu Ser Lys Gly Lys Asn Val Tyr Val
305 310 315 320
Lys Asn Asp Leu Ala Lys Thr Ala Met Ser Asn Ile Val Phe Glu Asn
325 330 335
Trp Arg Thr Phe Asp Asp Leu Leu Asn Gln Glu Tyr Asp Leu Ala Asn
340 345 350
Glu Asn Lys Lys Lys Asp Asp Lys Tyr Phe Glu Lys Arg Gln Lys Glu
355 360 365
Leu Lys Lys Asn Lys Ser Tyr Ser Leu Glu His Leu Cys Asn Leu Ser
370 375 380
Glu Asp Ser Cys Asn Leu Ile Glu Asn Tyr Ile His Gln Ile Ser Asp
385 390 395 400
Asp Ile Glu Asn Ile Ile Ile Asn Asn Glu Thr Phe Leu Arg Ile Val
405 410 415
Ile Asn Glu His Asp Arg Ser Arg Lys Leu Ala Lys Asn Arg Lys Ala
420 425 430
Val Lys Ala Ile Lys Asp Phe Leu Asp Ser Ile Lys Val Leu Glu Arg
435 440 445
Glu Leu Lys Leu Ile Asn Ser Ser Gly Gln Glu Leu Glu Lys Asp Leu
450 455 460
Ile Val Tyr Ser Ala His Glu Glu Leu Leu Val Glu Leu Lys Gln Val
465 470 475 480
Asp Ser Leu Tyr Asn Met Thr Arg Asn Tyr Leu Thr Lys Lys Pro Phe
485 490 495
Ser Thr Glu Lys Val Lys Leu Asn Phe Asn Arg Ser Thr Leu Leu Asn
500 505 510
Gly Trp Asp Arg Asn Lys Glu Thr Asp Asn Leu Gly Val Leu Leu Leu
515 520 525
Lys Asp Gly Lys Tyr Tyr Leu Gly Ile Met Asn Thr Ser Ala Asn Lys
530 535 540
Ala Phe Val Asn Pro Pro Val Ala Lys Thr Glu Lys Val Phe Lys Lys
545 550 555 560
Val Asp Tyr Lys Leu Leu Pro Val Pro Asn Gln Met Leu Pro Lys Val
565 570 575
Phe Phe Ala Lys Ser Asn Ile Asp Phe Tyr Asn Pro Ser Ser Glu Ile
580 585 590
Tyr Ser Asn Tyr Lys Lys Gly Thr His Lys Lys Gly Asn Met Phe Ser
595 600 605
Leu Glu Asp Cys His Asn Leu Ile Asp Phe Phe Lys Glu Ser Ile Ser
610 615 620
Lys His Glu Asp Trp Ser Lys Phe Gly Phe Lys Phe Ser Asp Thr Ala
625 630 635 640
Ser Tyr Asn Asp Ile Ser Glu Phe Tyr Arg Glu Val Glu Lys Gln Gly
645 650 655
Tyr Lys Leu Thr Tyr Thr Asp Ile Asp Glu Thr Tyr Ile Asn Asp Leu
660 665 670
Ile Glu Arg Asn Glu Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe
675 680 685
Ser Met Tyr Ser Lys Gly Lys Leu Asn Leu His Thr Leu Tyr Phe Met
690 695 700
Met Leu Phe Asp Gln Arg Asn Ile Asp Asp Val Val Tyr Lys Leu Asn
705 710 715 720
Gly Glu Ala Glu Val Phe Tyr Arg Pro Ala Ser Ile Ser Glu Asp Glu
725 730 735
Leu Ile Ile His Lys Ala Gly Glu Glu Ile Lys Asn Lys Asn Pro Asn
740 745 750
Arg Ala Arg Thr Lys Glu Thr Ser Thr Phe Ser Tyr Asp Ile Val Lys
755 760 765
Asp Lys Arg Tyr Ser Lys Asp Lys Phe Thr Leu His Ile Pro Ile Thr
770 775 780
Met Asn Phe Gly Val Asp Glu Val Lys Arg Phe Asn Asp Ala Val Asn
785 790 795 800
Ser Ala Ile Arg Ile Asp Glu Asn Val Asn Val Ile Gly Ile Asp Arg
805 810 815
Gly Glu Arg Asn Leu Leu Tyr Val Val Val Ile Asp Ser Lys Gly Asn
820 825 830
Ile Leu Glu Gln Ile Ser Leu Asn Ser Ile Ile Asn Lys Glu Tyr Asp
835 840 845
Ile Glu Thr Asp Tyr His Ala Leu Leu Asp Glu Arg Glu Gly Gly Arg
850 855 860
Asp Lys Ala Arg Lys Asp Trp Asn Thr Val Glu Asn Ile Arg Asp Leu
865 870 875 880
Lys Ala Gly Tyr Leu Ser Gln Val Val Asn Val Val Ala Lys Leu Val
885 890 895
Leu Lys Tyr Asn Ala Ile Ile Cys Leu Glu Asp Leu Asn Phe Gly Phe
900 905 910
Lys Arg Gly Arg Gln Lys Val Glu Lys Gln Val Tyr Gln Lys Phe Glu
915 920 925
Lys Met Leu Ile Asp Lys Leu Asn Tyr Leu Val Ile Asp Lys Ser Arg
930 935 940
Glu Gln Thr Ser Pro Lys Glu Leu Gly Gly Ala Leu Asn Ala Leu Gln
945 950 955 960
Leu Thr Ser Lys Phe Lys Ser Phe Lys Glu Leu Gly Lys Gln Ser Gly
965 970 975
Val Ile Tyr Tyr Val Pro Ala Tyr Leu Thr Ser Lys Ile Asp Pro Thr
980 985 990
Thr Gly Phe Ala Asn Leu Phe Tyr Met Lys Cys Glu Asn Val Glu Lys
995 1000 1005
Ser Lys Arg Phe Phe Asp Gly Phe Asp Phe Ile Arg Phe Asn Ala
1010 1015 1020
Leu Glu Asn Val Phe Glu Phe Gly Phe Asp Tyr Arg Ser Phe Thr
1025 1030 1035
Gln Arg Ala Cys Gly Ile Asn Ser Lys Trp Thr Val Cys Thr Asn
1040 1045 1050
Gly Glu Arg Ile Ile Lys Tyr Arg Asn Pro Asp Lys Asn Asn Met
1055 1060 1065
Phe Asp Glu Lys Val Val Val Val Thr Asp Glu Met Lys Asn Leu
1070 1075 1080
Phe Glu Gln Tyr Lys Ile Pro Tyr Glu Asp Gly Arg Asn Val Lys
1085 1090 1095
Asp Met Ile Ile Ser Asn Glu Glu Ala Glu Phe Tyr Arg Arg Leu
1100 1105 1110
Tyr Arg Leu Leu Gln Gln Thr Leu Gln Met Arg Asn Ser Thr Ser
1115 1120 1125
Asp Gly Thr Arg Asp Tyr Ile Ile Ser Pro Val Lys Asn Lys Arg
1130 1135 1140
Glu Ala Tyr Phe Asn Ser Glu Leu Ser Asp Gly Ser Val Pro Lys
1145 1150 1155
Asp Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala Arg Lys Gly Leu
1160 1165 1170
Trp Val Leu Glu Gln Ile Arg Gln Lys Ser Glu Gly Glu Lys Ile
1175 1180 1185
Asn Leu Ala Met Thr Asn Ala Glu Trp Leu Glu Tyr Ala Gln Thr
1190 1195 1200
His Leu Leu Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala
1205 1210 1215
Lys Lys Lys Lys Gly Ser Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
1220 1225 1230
Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Tyr Pro Tyr Asp Val Pro
1235 1240 1245
Asp Tyr Ala
1250
<210> 15
<211> 1283
<212> PRT
<213> 白蚁产甲烷菌暂定种(Candidatus Methanoplasma termitum)
<220>
<221> MISC_FEATURE
<222> (1)..(1283)
<223> CMtCpf1; pY012
<400> 15
Met Asn Asn Tyr Asp Glu Phe Thr Lys Leu Tyr Pro Ile Gln Lys Thr
1 5 10 15
Ile Arg Phe Glu Leu Lys Pro Gln Gly Arg Thr Met Glu His Leu Glu
20 25 30
Thr Phe Asn Phe Phe Glu Glu Asp Arg Asp Arg Ala Glu Lys Tyr Lys
35 40 45
Ile Leu Lys Glu Ala Ile Asp Glu Tyr His Lys Lys Phe Ile Asp Glu
50 55 60
His Leu Thr Asn Met Ser Leu Asp Trp Asn Ser Leu Lys Gln Ile Ser
65 70 75 80
Glu Lys Tyr Tyr Lys Ser Arg Glu Glu Lys Asp Lys Lys Val Phe Leu
85 90 95
Ser Glu Gln Lys Arg Met Arg Gln Glu Ile Val Ser Glu Phe Lys Lys
100 105 110
Asp Asp Arg Phe Lys Asp Leu Phe Ser Lys Lys Leu Phe Ser Glu Leu
115 120 125
Leu Lys Glu Glu Ile Tyr Lys Lys Gly Asn His Gln Glu Ile Asp Ala
130 135 140
Leu Lys Ser Phe Asp Lys Phe Ser Gly Tyr Phe Ile Gly Leu His Glu
145 150 155 160
Asn Arg Lys Asn Met Tyr Ser Asp Gly Asp Glu Ile Thr Ala Ile Ser
165 170 175
Asn Arg Ile Val Asn Glu Asn Phe Pro Lys Phe Leu Asp Asn Leu Gln
180 185 190
Lys Tyr Gln Glu Ala Arg Lys Lys Tyr Pro Glu Trp Ile Ile Lys Ala
195 200 205
Glu Ser Ala Leu Val Ala His Asn Ile Lys Met Asp Glu Val Phe Ser
210 215 220
Leu Glu Tyr Phe Asn Lys Val Leu Asn Gln Glu Gly Ile Gln Arg Tyr
225 230 235 240
Asn Leu Ala Leu Gly Gly Tyr Val Thr Lys Ser Gly Glu Lys Met Met
245 250 255
Gly Leu Asn Asp Ala Leu Asn Leu Ala His Gln Ser Glu Lys Ser Ser
260 265 270
Lys Gly Arg Ile His Met Thr Pro Leu Phe Lys Gln Ile Leu Ser Glu
275 280 285
Lys Glu Ser Phe Ser Tyr Ile Pro Asp Val Phe Thr Glu Asp Ser Gln
290 295 300
Leu Leu Pro Ser Ile Gly Gly Phe Phe Ala Gln Ile Glu Asn Asp Lys
305 310 315 320
Asp Gly Asn Ile Phe Asp Arg Ala Leu Glu Leu Ile Ser Ser Tyr Ala
325 330 335
Glu Tyr Asp Thr Glu Arg Ile Tyr Ile Arg Gln Ala Asp Ile Asn Arg
340 345 350
Val Ser Asn Val Ile Phe Gly Glu Trp Gly Thr Leu Gly Gly Leu Met
355 360 365
Arg Glu Tyr Lys Ala Asp Ser Ile Asn Asp Ile Asn Leu Glu Arg Thr
370 375 380
Cys Lys Lys Val Asp Lys Trp Leu Asp Ser Lys Glu Phe Ala Leu Ser
385 390 395 400
Asp Val Leu Glu Ala Ile Lys Arg Thr Gly Asn Asn Asp Ala Phe Asn
405 410 415
Glu Tyr Ile Ser Lys Met Arg Thr Ala Arg Glu Lys Ile Asp Ala Ala
420 425 430
Arg Lys Glu Met Lys Phe Ile Ser Glu Lys Ile Ser Gly Asp Glu Glu
435 440 445
Ser Ile His Ile Ile Lys Thr Leu Leu Asp Ser Val Gln Gln Phe Leu
450 455 460
His Phe Phe Asn Leu Phe Lys Ala Arg Gln Asp Ile Pro Leu Asp Gly
465 470 475 480
Ala Phe Tyr Ala Glu Phe Asp Glu Val His Ser Lys Leu Phe Ala Ile
485 490 495
Val Pro Leu Tyr Asn Lys Val Arg Asn Tyr Leu Thr Lys Asn Asn Leu
500 505 510
Asn Thr Lys Lys Ile Lys Leu Asn Phe Lys Asn Pro Thr Leu Ala Asn
515 520 525
Gly Trp Asp Gln Asn Lys Val Tyr Asp Tyr Ala Ser Leu Ile Phe Leu
530 535 540
Arg Asp Gly Asn Tyr Tyr Leu Gly Ile Ile Asn Pro Lys Arg Lys Lys
545 550 555 560
Asn Ile Lys Phe Glu Gln Gly Ser Gly Asn Gly Pro Phe Tyr Arg Lys
565 570 575
Met Val Tyr Lys Gln Ile Pro Gly Pro Asn Lys Asn Leu Pro Arg Val
580 585 590
Phe Leu Thr Ser Thr Lys Gly Lys Lys Glu Tyr Lys Pro Ser Lys Glu
595 600 605
Ile Ile Glu Gly Tyr Glu Ala Asp Lys His Ile Arg Gly Asp Lys Phe
610 615 620
Asp Leu Asp Phe Cys His Lys Leu Ile Asp Phe Phe Lys Glu Ser Ile
625 630 635 640
Glu Lys His Lys Asp Trp Ser Lys Phe Asn Phe Tyr Phe Ser Pro Thr
645 650 655
Glu Ser Tyr Gly Asp Ile Ser Glu Phe Tyr Leu Asp Val Glu Lys Gln
660 665 670
Gly Tyr Arg Met His Phe Glu Asn Ile Ser Ala Glu Thr Ile Asp Glu
675 680 685
Tyr Val Glu Lys Gly Asp Leu Phe Leu Phe Gln Ile Tyr Asn Lys Asp
690 695 700
Phe Val Lys Ala Ala Thr Gly Lys Lys Asp Met His Thr Ile Tyr Trp
705 710 715 720
Asn Ala Ala Phe Ser Pro Glu Asn Leu Gln Asp Val Val Val Lys Leu
725 730 735
Asn Gly Glu Ala Glu Leu Phe Tyr Arg Asp Lys Ser Asp Ile Lys Glu
740 745 750
Ile Val His Arg Glu Gly Glu Ile Leu Val Asn Arg Thr Tyr Asn Gly
755 760 765
Arg Thr Pro Val Pro Asp Lys Ile His Lys Lys Leu Thr Asp Tyr His
770 775 780
Asn Gly Arg Thr Lys Asp Leu Gly Glu Ala Lys Glu Tyr Leu Asp Lys
785 790 795 800
Val Arg Tyr Phe Lys Ala His Tyr Asp Ile Thr Lys Asp Arg Arg Tyr
805 810 815
Leu Asn Asp Lys Ile Tyr Phe His Val Pro Leu Thr Leu Asn Phe Lys
820 825 830
Ala Asn Gly Lys Lys Asn Leu Asn Lys Met Val Ile Glu Lys Phe Leu
835 840 845
Ser Asp Glu Lys Ala His Ile Ile Gly Ile Asp Arg Gly Glu Arg Asn
850 855 860
Leu Leu Tyr Tyr Ser Ile Ile Asp Arg Ser Gly Lys Ile Ile Asp Gln
865 870 875 880
Gln Ser Leu Asn Val Ile Asp Gly Phe Asp Tyr Arg Glu Lys Leu Asn
885 890 895
Gln Arg Glu Ile Glu Met Lys Asp Ala Arg Gln Ser Trp Asn Ala Ile
900 905 910
Gly Lys Ile Lys Asp Leu Lys Glu Gly Tyr Leu Ser Lys Ala Val His
915 920 925
Glu Ile Thr Lys Met Ala Ile Gln Tyr Asn Ala Ile Val Val Met Glu
930 935 940
Glu Leu Asn Tyr Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln
945 950 955 960
Ile Tyr Gln Lys Phe Glu Asn Met Leu Ile Asp Lys Met Asn Tyr Leu
965 970 975
Val Phe Lys Asp Ala Pro Asp Glu Ser Pro Gly Gly Val Leu Asn Ala
980 985 990
Tyr Gln Leu Thr Asn Pro Leu Glu Ser Phe Ala Lys Leu Gly Lys Gln
995 1000 1005
Thr Gly Ile Leu Phe Tyr Val Pro Ala Ala Tyr Thr Ser Lys Ile
1010 1015 1020
Asp Pro Thr Thr Gly Phe Val Asn Leu Phe Asn Thr Ser Ser Lys
1025 1030 1035
Thr Asn Ala Gln Glu Arg Lys Glu Phe Leu Gln Lys Phe Glu Ser
1040 1045 1050
Ile Ser Tyr Ser Ala Lys Asp Gly Gly Ile Phe Ala Phe Ala Phe
1055 1060 1065
Asp Tyr Arg Lys Phe Gly Thr Ser Lys Thr Asp His Lys Asn Val
1070 1075 1080
Trp Thr Ala Tyr Thr Asn Gly Glu Arg Met Arg Tyr Ile Lys Glu
1085 1090 1095
Lys Lys Arg Asn Glu Leu Phe Asp Pro Ser Lys Glu Ile Lys Glu
1100 1105 1110
Ala Leu Thr Ser Ser Gly Ile Lys Tyr Asp Gly Gly Gln Asn Ile
1115 1120 1125
Leu Pro Asp Ile Leu Arg Ser Asn Asn Asn Gly Leu Ile Tyr Thr
1130 1135 1140
Met Tyr Ser Ser Phe Ile Ala Ala Ile Gln Met Arg Val Tyr Asp
1145 1150 1155
Gly Lys Glu Asp Tyr Ile Ile Ser Pro Ile Lys Asn Ser Lys Gly
1160 1165 1170
Glu Phe Phe Arg Thr Asp Pro Lys Arg Arg Glu Leu Pro Ile Asp
1175 1180 1185
Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala Leu Arg Gly Glu Leu
1190 1195 1200
Thr Met Arg Ala Ile Ala Glu Lys Phe Asp Pro Asp Ser Glu Lys
1205 1210 1215
Met Ala Lys Leu Glu Leu Lys His Lys Asp Trp Phe Glu Phe Met
1220 1225 1230
Gln Thr Arg Gly Asp Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly
1235 1240 1245
Gln Ala Lys Lys Lys Lys Gly Ser Tyr Pro Tyr Asp Val Pro Asp
1250 1255 1260
Tyr Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Tyr Pro Tyr Asp
1265 1270 1275
Val Pro Asp Tyr Ala
1280
<210> 16
<211> 1327
<212> PRT
<213> 挑剔真杆菌
<220>
<221> MISC_FEATURE
<222> (1)..(1327)
<223> EeCpf1; pY013
<400> 16
Met Asn Gly Asn Arg Ser Ile Val Tyr Arg Glu Phe Val Gly Val Ile
1 5 10 15
Pro Val Ala Lys Thr Leu Arg Asn Glu Leu Arg Pro Val Gly His Thr
20 25 30
Gln Glu His Ile Ile Gln Asn Gly Leu Ile Gln Glu Asp Glu Leu Arg
35 40 45
Gln Glu Lys Ser Thr Glu Leu Lys Asn Ile Met Asp Asp Tyr Tyr Arg
50 55 60
Glu Tyr Ile Asp Lys Ser Leu Ser Gly Val Thr Asp Leu Asp Phe Thr
65 70 75 80
Leu Leu Phe Glu Leu Met Asn Leu Val Gln Ser Ser Pro Ser Lys Asp
85 90 95
Asn Lys Lys Ala Leu Glu Lys Glu Gln Ser Lys Met Arg Glu Gln Ile
100 105 110
Cys Thr His Leu Gln Ser Asp Ser Asn Tyr Lys Asn Ile Phe Asn Ala
115 120 125
Lys Leu Leu Lys Glu Ile Leu Pro Asp Phe Ile Lys Asn Tyr Asn Gln
130 135 140
Tyr Asp Val Lys Asp Lys Ala Gly Lys Leu Glu Thr Leu Ala Leu Phe
145 150 155 160
Asn Gly Phe Ser Thr Tyr Phe Thr Asp Phe Phe Glu Lys Arg Lys Asn
165 170 175
Val Phe Thr Lys Glu Ala Val Ser Thr Ser Ile Ala Tyr Arg Ile Val
180 185 190
His Glu Asn Ser Leu Ile Phe Leu Ala Asn Met Thr Ser Tyr Lys Lys
195 200 205
Ile Ser Glu Lys Ala Leu Asp Glu Ile Glu Val Ile Glu Lys Asn Asn
210 215 220
Gln Asp Lys Met Gly Asp Trp Glu Leu Asn Gln Ile Phe Asn Pro Asp
225 230 235 240
Phe Tyr Asn Met Val Leu Ile Gln Ser Gly Ile Asp Phe Tyr Asn Glu
245 250 255
Ile Cys Gly Val Val Asn Ala His Met Asn Leu Tyr Cys Gln Gln Thr
260 265 270
Lys Asn Asn Tyr Asn Leu Phe Lys Met Arg Lys Leu His Lys Gln Ile
275 280 285
Leu Ala Tyr Thr Ser Thr Ser Phe Glu Val Pro Lys Met Phe Glu Asp
290 295 300
Asp Met Ser Val Tyr Asn Ala Val Asn Ala Phe Ile Asp Glu Thr Glu
305 310 315 320
Lys Gly Asn Ile Ile Gly Lys Leu Lys Asp Ile Val Asn Lys Tyr Asp
325 330 335
Glu Leu Asp Glu Lys Arg Ile Tyr Ile Ser Lys Asp Phe Tyr Glu Thr
340 345 350
Leu Ser Cys Phe Met Ser Gly Asn Trp Asn Leu Ile Thr Gly Cys Val
355 360 365
Glu Asn Phe Tyr Asp Glu Asn Ile His Ala Lys Gly Lys Ser Lys Glu
370 375 380
Glu Lys Val Lys Lys Ala Val Lys Glu Asp Lys Tyr Lys Ser Ile Asn
385 390 395 400
Asp Val Asn Asp Leu Val Glu Lys Tyr Ile Asp Glu Lys Glu Arg Asn
405 410 415
Glu Phe Lys Asn Ser Asn Ala Lys Gln Tyr Ile Arg Glu Ile Ser Asn
420 425 430
Ile Ile Thr Asp Thr Glu Thr Ala His Leu Glu Tyr Asp Asp His Ile
435 440 445
Ser Leu Ile Glu Ser Glu Glu Lys Ala Asp Glu Met Lys Lys Arg Leu
450 455 460
Asp Met Tyr Met Asn Met Tyr His Trp Ala Lys Ala Phe Ile Val Asp
465 470 475 480
Glu Val Leu Asp Arg Asp Glu Met Phe Tyr Ser Asp Ile Asp Asp Ile
485 490 495
Tyr Asn Ile Leu Glu Asn Ile Val Pro Leu Tyr Asn Arg Val Arg Asn
500 505 510
Tyr Val Thr Gln Lys Pro Tyr Asn Ser Lys Lys Ile Lys Leu Asn Phe
515 520 525
Gln Ser Pro Thr Leu Ala Asn Gly Trp Ser Gln Ser Lys Glu Phe Asp
530 535 540
Asn Asn Ala Ile Ile Leu Ile Arg Asp Asn Lys Tyr Tyr Leu Ala Ile
545 550 555 560
Phe Asn Ala Lys Asn Lys Pro Asp Lys Lys Ile Ile Gln Gly Asn Ser
565 570 575
Asp Lys Lys Asn Asp Asn Asp Tyr Lys Lys Met Val Tyr Asn Leu Leu
580 585 590
Pro Gly Ala Asn Lys Met Leu Pro Lys Val Phe Leu Ser Lys Lys Gly
595 600 605
Ile Glu Thr Phe Lys Pro Ser Asp Tyr Ile Ile Ser Gly Tyr Asn Ala
610 615 620
His Lys His Ile Lys Thr Ser Glu Asn Phe Asp Ile Ser Phe Cys Arg
625 630 635 640
Asp Leu Ile Asp Tyr Phe Lys Asn Ser Ile Glu Lys His Ala Glu Trp
645 650 655
Arg Lys Tyr Glu Phe Lys Phe Ser Ala Thr Asp Ser Tyr Ser Asp Ile
660 665 670
Ser Glu Phe Tyr Arg Glu Val Glu Met Gln Gly Tyr Arg Ile Asp Trp
675 680 685
Thr Tyr Ile Ser Glu Ala Asp Ile Asn Lys Leu Asp Glu Glu Gly Lys
690 695 700
Ile Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ala Glu Asn Ser Thr
705 710 715 720
Gly Lys Glu Asn Leu His Thr Met Tyr Phe Lys Asn Ile Phe Ser Glu
725 730 735
Glu Asn Leu Lys Asp Ile Ile Ile Lys Leu Asn Gly Gln Ala Glu Leu
740 745 750
Phe Tyr Arg Arg Ala Ser Val Lys Asn Pro Val Lys His Lys Lys Asp
755 760 765
Ser Val Leu Val Asn Lys Thr Tyr Lys Asn Gln Leu Asp Asn Gly Asp
770 775 780
Val Val Arg Ile Pro Ile Pro Asp Asp Ile Tyr Asn Glu Ile Tyr Lys
785 790 795 800
Met Tyr Asn Gly Tyr Ile Lys Glu Ser Asp Leu Ser Glu Ala Ala Lys
805 810 815
Glu Tyr Leu Asp Lys Val Glu Val Arg Thr Ala Gln Lys Asp Ile Val
820 825 830
Lys Asp Tyr Arg Tyr Thr Val Asp Lys Tyr Phe Ile His Thr Pro Ile
835 840 845
Thr Ile Asn Tyr Lys Val Thr Ala Arg Asn Asn Val Asn Asp Met Val
850 855 860
Val Lys Tyr Ile Ala Gln Asn Asp Asp Ile His Val Ile Gly Ile Asp
865 870 875 880
Arg Gly Glu Arg Asn Leu Ile Tyr Ile Ser Val Ile Asp Ser His Gly
885 890 895
Asn Ile Val Lys Gln Lys Ser Tyr Asn Ile Leu Asn Asn Tyr Asp Tyr
900 905 910
Lys Lys Lys Leu Val Glu Lys Glu Lys Thr Arg Glu Tyr Ala Arg Lys
915 920 925
Asn Trp Lys Ser Ile Gly Asn Ile Lys Glu Leu Lys Glu Gly Tyr Ile
930 935 940
Ser Gly Val Val His Glu Ile Ala Met Leu Ile Val Glu Tyr Asn Ala
945 950 955 960
Ile Ile Ala Met Glu Asp Leu Asn Tyr Gly Phe Lys Arg Gly Arg Phe
965 970 975
Lys Val Glu Arg Gln Val Tyr Gln Lys Phe Glu Ser Met Leu Ile Asn
980 985 990
Lys Leu Asn Tyr Phe Ala Ser Lys Glu Lys Ser Val Asp Glu Pro Gly
995 1000 1005
Gly Leu Leu Lys Gly Tyr Gln Leu Thr Tyr Val Pro Asp Asn Ile
1010 1015 1020
Lys Asn Leu Gly Lys Gln Cys Gly Val Ile Phe Tyr Val Pro Ala
1025 1030 1035
Ala Phe Thr Ser Lys Ile Asp Pro Ser Thr Gly Phe Ile Ser Ala
1040 1045 1050
Phe Asn Phe Lys Ser Ile Ser Thr Asn Ala Ser Arg Lys Gln Phe
1055 1060 1065
Phe Met Gln Phe Asp Glu Ile Arg Tyr Cys Ala Glu Lys Asp Met
1070 1075 1080
Phe Ser Phe Gly Phe Asp Tyr Asn Asn Phe Asp Thr Tyr Asn Ile
1085 1090 1095
Thr Met Gly Lys Thr Gln Trp Thr Val Tyr Thr Asn Gly Glu Arg
1100 1105 1110
Leu Gln Ser Glu Phe Asn Asn Ala Arg Arg Thr Gly Lys Thr Lys
1115 1120 1125
Ser Ile Asn Leu Thr Glu Thr Ile Lys Leu Leu Leu Glu Asp Asn
1130 1135 1140
Glu Ile Asn Tyr Ala Asp Gly His Asp Ile Arg Ile Asp Met Glu
1145 1150 1155
Lys Met Asp Glu Asp Lys Lys Ser Glu Phe Phe Ala Gln Leu Leu
1160 1165 1170
Ser Leu Tyr Lys Leu Thr Val Gln Met Arg Asn Ser Tyr Thr Glu
1175 1180 1185
Ala Glu Glu Gln Glu Asn Gly Ile Ser Tyr Asp Lys Ile Ile Ser
1190 1195 1200
Pro Val Ile Asn Asp Glu Gly Glu Phe Phe Asp Ser Asp Asn Tyr
1205 1210 1215
Lys Glu Ser Asp Asp Lys Glu Cys Lys Met Pro Lys Asp Ala Asp
1220 1225 1230
Ala Asn Gly Ala Tyr Cys Ile Ala Leu Lys Gly Leu Tyr Glu Val
1235 1240 1245
Leu Lys Ile Lys Ser Glu Trp Thr Glu Asp Gly Phe Asp Arg Asn
1250 1255 1260
Cys Leu Lys Leu Pro His Ala Glu Trp Leu Asp Phe Ile Gln Asn
1265 1270 1275
Lys Arg Tyr Glu Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln
1280 1285 1290
Ala Lys Lys Lys Lys Gly Ser Tyr Pro Tyr Asp Val Pro Asp Tyr
1295 1300 1305
Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Tyr Pro Tyr Asp Val
1310 1315 1320
Pro Asp Tyr Ala
1325
<210> 17
<211> 1418
<212> PRT
<213> 牛眼莫拉氏菌237
<220>
<221> MISC_FEATURE
<222> (1)..(1418)
<223> MbCpf1; pY014
<400> 17
Met Leu Phe Gln Asp Phe Thr His Leu Tyr Pro Leu Ser Lys Thr Val
1 5 10 15
Arg Phe Glu Leu Lys Pro Ile Asp Arg Thr Leu Glu His Ile His Ala
20 25 30
Lys Asn Phe Leu Ser Gln Asp Glu Thr Met Ala Asp Met His Gln Lys
35 40 45
Val Lys Val Ile Leu Asp Asp Tyr His Arg Asp Phe Ile Ala Asp Met
50 55 60
Met Gly Glu Val Lys Leu Thr Lys Leu Ala Glu Phe Tyr Asp Val Tyr
65 70 75 80
Leu Lys Phe Arg Lys Asn Pro Lys Asp Asp Glu Leu Gln Lys Gln Leu
85 90 95
Lys Asp Leu Gln Ala Val Leu Arg Lys Glu Ile Val Lys Pro Ile Gly
100 105 110
Asn Gly Gly Lys Tyr Lys Ala Gly Tyr Asp Arg Leu Phe Gly Ala Lys
115 120 125
Leu Phe Lys Asp Gly Lys Glu Leu Gly Asp Leu Ala Lys Phe Val Ile
130 135 140
Ala Gln Glu Gly Glu Ser Ser Pro Lys Leu Ala His Leu Ala His Phe
145 150 155 160
Glu Lys Phe Ser Thr Tyr Phe Thr Gly Phe His Asp Asn Arg Lys Asn
165 170 175
Met Tyr Ser Asp Glu Asp Lys His Thr Ala Ile Ala Tyr Arg Leu Ile
180 185 190
His Glu Asn Leu Pro Arg Phe Ile Asp Asn Leu Gln Ile Leu Thr Thr
195 200 205
Ile Lys Gln Lys His Ser Ala Leu Tyr Asp Gln Ile Ile Asn Glu Leu
210 215 220
Thr Ala Ser Gly Leu Asp Val Ser Leu Ala Ser His Leu Asp Gly Tyr
225 230 235 240
His Lys Leu Leu Thr Gln Glu Gly Ile Thr Ala Tyr Asn Thr Leu Leu
245 250 255
Gly Gly Ile Ser Gly Glu Ala Gly Ser Pro Lys Ile Gln Gly Ile Asn
260 265 270
Glu Leu Ile Asn Ser His His Asn Gln His Cys His Lys Ser Glu Arg
275 280 285
Ile Ala Lys Leu Arg Pro Leu His Lys Gln Ile Leu Ser Asp Gly Met
290 295 300
Ser Val Ser Phe Leu Pro Ser Lys Phe Ala Asp Asp Ser Glu Met Cys
305 310 315 320
Gln Ala Val Asn Glu Phe Tyr Arg His Tyr Ala Asp Val Phe Ala Lys
325 330 335
Val Gln Ser Leu Phe Asp Gly Phe Asp Asp His Gln Lys Asp Gly Ile
340 345 350
Tyr Val Glu His Lys Asn Leu Asn Glu Leu Ser Lys Gln Ala Phe Gly
355 360 365
Asp Phe Ala Leu Leu Gly Arg Val Leu Asp Gly Tyr Tyr Val Asp Val
370 375 380
Val Asn Pro Glu Phe Asn Glu Arg Phe Ala Lys Ala Lys Thr Asp Asn
385 390 395 400
Ala Lys Ala Lys Leu Thr Lys Glu Lys Asp Lys Phe Ile Lys Gly Val
405 410 415
His Ser Leu Ala Ser Leu Glu Gln Ala Ile Glu His Tyr Thr Ala Arg
420 425 430
His Asp Asp Glu Ser Val Gln Ala Gly Lys Leu Gly Gln Tyr Phe Lys
435 440 445
His Gly Leu Ala Gly Val Asp Asn Pro Ile Gln Lys Ile His Asn Asn
450 455 460
His Ser Thr Ile Lys Gly Phe Leu Glu Arg Glu Arg Pro Ala Gly Glu
465 470 475 480
Arg Ala Leu Pro Lys Ile Lys Ser Gly Lys Asn Pro Glu Met Thr Gln
485 490 495
Leu Arg Gln Leu Lys Glu Leu Leu Asp Asn Ala Leu Asn Val Ala His
500 505 510
Phe Ala Lys Leu Leu Thr Thr Lys Thr Thr Leu Asp Asn Gln Asp Gly
515 520 525
Asn Phe Tyr Gly Glu Phe Gly Val Leu Tyr Asp Glu Leu Ala Lys Ile
530 535 540
Pro Thr Leu Tyr Asn Lys Val Arg Asp Tyr Leu Ser Gln Lys Pro Phe
545 550 555 560
Ser Thr Glu Lys Tyr Lys Leu Asn Phe Gly Asn Pro Thr Leu Leu Asn
565 570 575
Gly Trp Asp Leu Asn Lys Glu Lys Asp Asn Phe Gly Val Ile Leu Gln
580 585 590
Lys Asp Gly Cys Tyr Tyr Leu Ala Leu Leu Asp Lys Ala His Lys Lys
595 600 605
Val Phe Asp Asn Ala Pro Asn Thr Gly Lys Ser Ile Tyr Gln Lys Met
610 615 620
Ile Tyr Lys Tyr Leu Glu Val Arg Lys Gln Phe Pro Lys Val Phe Phe
625 630 635 640
Ser Lys Glu Ala Ile Ala Ile Asn Tyr His Pro Ser Lys Glu Leu Val
645 650 655
Glu Ile Lys Asp Lys Gly Arg Gln Arg Ser Asp Asp Glu Arg Leu Lys
660 665 670
Leu Tyr Arg Phe Ile Leu Glu Cys Leu Lys Ile His Pro Lys Tyr Asp
675 680 685
Lys Lys Phe Glu Gly Ala Ile Gly Asp Ile Gln Leu Phe Lys Lys Asp
690 695 700
Lys Lys Gly Arg Glu Val Pro Ile Ser Glu Lys Asp Leu Phe Asp Lys
705 710 715 720
Ile Asn Gly Ile Phe Ser Ser Lys Pro Lys Leu Glu Met Glu Asp Phe
725 730 735
Phe Ile Gly Glu Phe Lys Arg Tyr Asn Pro Ser Gln Asp Leu Val Asp
740 745 750
Gln Tyr Asn Ile Tyr Lys Lys Ile Asp Ser Asn Asp Asn Arg Lys Lys
755 760 765
Glu Asn Phe Tyr Asn Asn His Pro Lys Phe Lys Lys Asp Leu Val Arg
770 775 780
Tyr Tyr Tyr Glu Ser Met Cys Lys His Glu Glu Trp Glu Glu Ser Phe
785 790 795 800
Glu Phe Ser Lys Lys Leu Gln Asp Ile Gly Cys Tyr Val Asp Val Asn
805 810 815
Glu Leu Phe Thr Glu Ile Glu Thr Arg Arg Leu Asn Tyr Lys Ile Ser
820 825 830
Phe Cys Asn Ile Asn Ala Asp Tyr Ile Asp Glu Leu Val Glu Gln Gly
835 840 845
Gln Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Pro Lys Ala
850 855 860
His Gly Lys Pro Asn Leu His Thr Leu Tyr Phe Lys Ala Leu Phe Ser
865 870 875 880
Glu Asp Asn Leu Ala Asp Pro Ile Tyr Lys Leu Asn Gly Glu Ala Gln
885 890 895
Ile Phe Tyr Arg Lys Ala Ser Leu Asp Met Asn Glu Thr Thr Ile His
900 905 910
Arg Ala Gly Glu Val Leu Glu Asn Lys Asn Pro Asp Asn Pro Lys Lys
915 920 925
Arg Gln Phe Val Tyr Asp Ile Ile Lys Asp Lys Arg Tyr Thr Gln Asp
930 935 940
Lys Phe Met Leu His Val Pro Ile Thr Met Asn Phe Gly Val Gln Gly
945 950 955 960
Met Thr Ile Lys Glu Phe Asn Lys Lys Val Asn Gln Ser Ile Gln Gln
965 970 975
Tyr Asp Glu Val Asn Val Ile Gly Ile Asp Arg Gly Glu Arg His Leu
980 985 990
Leu Tyr Leu Thr Val Ile Asn Ser Lys Gly Glu Ile Leu Glu Gln Cys
995 1000 1005
Ser Leu Asn Asp Ile Thr Thr Ala Ser Ala Asn Gly Thr Gln Met
1010 1015 1020
Thr Thr Pro Tyr His Lys Ile Leu Asp Lys Arg Glu Ile Glu Arg
1025 1030 1035
Leu Asn Ala Arg Val Gly Trp Gly Glu Ile Glu Thr Ile Lys Glu
1040 1045 1050
Leu Lys Ser Gly Tyr Leu Ser His Val Val His Gln Ile Ser Gln
1055 1060 1065
Leu Met Leu Lys Tyr Asn Ala Ile Val Val Leu Glu Asp Leu Asn
1070 1075 1080
Phe Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Ile Tyr
1085 1090 1095
Gln Asn Phe Glu Asn Ala Leu Ile Lys Lys Leu Asn His Leu Val
1100 1105 1110
Leu Lys Asp Lys Ala Asp Asp Glu Ile Gly Ser Tyr Lys Asn Ala
1115 1120 1125
Leu Gln Leu Thr Asn Asn Phe Thr Asp Leu Lys Ser Ile Gly Lys
1130 1135 1140
Gln Thr Gly Phe Leu Phe Tyr Val Pro Ala Trp Asn Thr Ser Lys
1145 1150 1155
Ile Asp Pro Glu Thr Gly Phe Val Asp Leu Leu Lys Pro Arg Tyr
1160 1165 1170
Glu Asn Ile Ala Gln Ser Gln Ala Phe Phe Gly Lys Phe Asp Lys
1175 1180 1185
Ile Cys Tyr Asn Ala Asp Lys Asp Tyr Phe Glu Phe His Ile Asp
1190 1195 1200
Tyr Ala Lys Phe Thr Asp Lys Ala Lys Asn Ser Arg Gln Ile Trp
1205 1210 1215
Thr Ile Cys Ser His Gly Asp Lys Arg Tyr Val Tyr Asp Lys Thr
1220 1225 1230
Ala Asn Gln Asn Lys Gly Ala Ala Lys Gly Ile Asn Val Asn Asp
1235 1240 1245
Glu Leu Lys Ser Leu Phe Ala Arg His His Ile Asn Glu Lys Gln
1250 1255 1260
Pro Asn Leu Val Met Asp Ile Cys Gln Asn Asn Asp Lys Glu Phe
1265 1270 1275
His Lys Ser Leu Met Tyr Leu Leu Lys Thr Leu Leu Ala Leu Arg
1280 1285 1290
Tyr Ser Asn Ala Ser Ser Asp Glu Asp Phe Ile Leu Ser Pro Val
1295 1300 1305
Ala Asn Asp Glu Gly Val Phe Phe Asn Ser Ala Leu Ala Asp Asp
1310 1315 1320
Thr Gln Pro Gln Asn Ala Asp Ala Asn Gly Ala Tyr His Ile Ala
1325 1330 1335
Leu Lys Gly Leu Trp Leu Leu Asn Glu Leu Lys Asn Ser Asp Asp
1340 1345 1350
Leu Asn Lys Val Lys Leu Ala Ile Asp Asn Gln Thr Trp Leu Asn
1355 1360 1365
Phe Ala Gln Asn Arg Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly
1370 1375 1380
Gln Ala Lys Lys Lys Lys Gly Ser Tyr Pro Tyr Asp Val Pro Asp
1385 1390 1395
Tyr Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Tyr Pro Tyr Asp
1400 1405 1410
Val Pro Asp Tyr Ala
1415
<210> 18
<211> 1308
<212> PRT
<213> 稻田氏钩端螺旋体
<220>
<221> MISC_FEATURE
<222> (1)..(1308)
<223> LiCpf1; pY015
<400> 18
Met Glu Asp Tyr Ser Gly Phe Val Asn Ile Tyr Ser Ile Gln Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Lys Pro Val Gly Lys Thr Leu Glu His Ile Glu
20 25 30
Lys Lys Gly Phe Leu Lys Lys Asp Lys Ile Arg Ala Glu Asp Tyr Lys
35 40 45
Ala Val Lys Lys Ile Ile Asp Lys Tyr His Arg Ala Tyr Ile Glu Glu
50 55 60
Val Phe Asp Ser Val Leu His Gln Lys Lys Lys Lys Asp Lys Thr Arg
65 70 75 80
Phe Ser Thr Gln Phe Ile Lys Glu Ile Lys Glu Phe Ser Glu Leu Tyr
85 90 95
Tyr Lys Thr Glu Lys Asn Ile Pro Asp Lys Glu Arg Leu Glu Ala Leu
100 105 110
Ser Glu Lys Leu Arg Lys Met Leu Val Gly Ala Phe Lys Gly Glu Phe
115 120 125
Ser Glu Glu Val Ala Glu Lys Tyr Lys Asn Leu Phe Ser Lys Glu Leu
130 135 140
Ile Arg Asn Glu Ile Glu Lys Phe Cys Glu Thr Asp Glu Glu Arg Lys
145 150 155 160
Gln Val Ser Asn Phe Lys Ser Phe Thr Thr Tyr Phe Thr Gly Phe His
165 170 175
Ser Asn Arg Gln Asn Ile Tyr Ser Asp Glu Lys Lys Ser Thr Ala Ile
180 185 190
Gly Tyr Arg Ile Ile His Gln Asn Leu Pro Lys Phe Leu Asp Asn Leu
195 200 205
Lys Ile Ile Glu Ser Ile Gln Arg Arg Phe Lys Asp Phe Pro Trp Ser
210 215 220
Asp Leu Lys Lys Asn Leu Lys Lys Ile Asp Lys Asn Ile Lys Leu Thr
225 230 235 240
Glu Tyr Phe Ser Ile Asp Gly Phe Val Asn Val Leu Asn Gln Lys Gly
245 250 255
Ile Asp Ala Tyr Asn Thr Ile Leu Gly Gly Lys Ser Glu Glu Ser Gly
260 265 270
Glu Lys Ile Gln Gly Leu Asn Glu Tyr Ile Asn Leu Tyr Arg Gln Lys
275 280 285
Asn Asn Ile Asp Arg Lys Asn Leu Pro Asn Val Lys Ile Leu Phe Lys
290 295 300
Gln Ile Leu Gly Asp Arg Glu Thr Lys Ser Phe Ile Pro Glu Ala Phe
305 310 315 320
Pro Asp Asp Gln Ser Val Leu Asn Ser Ile Thr Glu Phe Ala Lys Tyr
325 330 335
Leu Lys Leu Asp Lys Lys Lys Lys Ser Ile Ile Ala Glu Leu Lys Lys
340 345 350
Phe Leu Ser Ser Phe Asn Arg Tyr Glu Leu Asp Gly Ile Tyr Leu Ala
355 360 365
Asn Asp Asn Ser Leu Ala Ser Ile Ser Thr Phe Leu Phe Asp Asp Trp
370 375 380
Ser Phe Ile Lys Lys Ser Val Ser Phe Lys Tyr Asp Glu Ser Val Gly
385 390 395 400
Asp Pro Lys Lys Lys Ile Lys Ser Pro Leu Lys Tyr Glu Lys Glu Lys
405 410 415
Glu Lys Trp Leu Lys Gln Lys Tyr Tyr Thr Ile Ser Phe Leu Asn Asp
420 425 430
Ala Ile Glu Ser Tyr Ser Lys Ser Gln Asp Glu Lys Arg Val Lys Ile
435 440 445
Arg Leu Glu Ala Tyr Phe Ala Glu Phe Lys Ser Lys Asp Asp Ala Lys
450 455 460
Lys Gln Phe Asp Leu Leu Glu Arg Ile Glu Glu Ala Tyr Ala Ile Val
465 470 475 480
Glu Pro Leu Leu Gly Ala Glu Tyr Pro Arg Asp Arg Asn Leu Lys Ala
485 490 495
Asp Lys Lys Glu Val Gly Lys Ile Lys Asp Phe Leu Asp Ser Ile Lys
500 505 510
Ser Leu Gln Phe Phe Leu Lys Pro Leu Leu Ser Ala Glu Ile Phe Asp
515 520 525
Glu Lys Asp Leu Gly Phe Tyr Asn Gln Leu Glu Gly Tyr Tyr Glu Glu
530 535 540
Ile Asp Ser Ile Gly His Leu Tyr Asn Lys Val Arg Asn Tyr Leu Thr
545 550 555 560
Gly Lys Ile Tyr Ser Lys Glu Lys Phe Lys Leu Asn Phe Glu Asn Ser
565 570 575
Thr Leu Leu Lys Gly Trp Asp Glu Asn Arg Glu Val Ala Asn Leu Cys
580 585 590
Val Ile Phe Arg Glu Asp Gln Lys Tyr Tyr Leu Gly Val Met Asp Lys
595 600 605
Glu Asn Asn Thr Ile Leu Ser Asp Ile Pro Lys Val Lys Pro Asn Glu
610 615 620
Leu Phe Tyr Glu Lys Met Val Tyr Lys Leu Ile Pro Thr Pro His Met
625 630 635 640
Gln Leu Pro Arg Ile Ile Phe Ser Ser Asp Asn Leu Ser Ile Tyr Asn
645 650 655
Pro Ser Lys Ser Ile Leu Lys Ile Arg Glu Ala Lys Ser Phe Lys Glu
660 665 670
Gly Lys Asn Phe Lys Leu Lys Asp Cys His Lys Phe Ile Asp Phe Tyr
675 680 685
Lys Glu Ser Ile Ser Lys Asn Glu Asp Trp Ser Arg Phe Asp Phe Lys
690 695 700
Phe Ser Lys Thr Ser Ser Tyr Glu Asn Ile Ser Glu Phe Tyr Arg Glu
705 710 715 720
Val Glu Arg Gln Gly Tyr Asn Leu Asp Phe Lys Lys Val Ser Lys Phe
725 730 735
Tyr Ile Asp Ser Leu Val Glu Asp Gly Lys Leu Tyr Leu Phe Gln Ile
740 745 750
Tyr Asn Lys Asp Phe Ser Ile Phe Ser Lys Gly Lys Pro Asn Leu His
755 760 765
Thr Ile Tyr Phe Arg Ser Leu Phe Ser Lys Glu Asn Leu Lys Asp Val
770 775 780
Cys Leu Lys Leu Asn Gly Glu Ala Glu Met Phe Phe Arg Lys Lys Ser
785 790 795 800
Ile Asn Tyr Asp Glu Lys Lys Lys Arg Glu Gly His His Pro Glu Leu
805 810 815
Phe Glu Lys Leu Lys Tyr Pro Ile Leu Lys Asp Lys Arg Tyr Ser Glu
820 825 830
Asp Lys Phe Gln Phe His Leu Pro Ile Ser Leu Asn Phe Lys Ser Lys
835 840 845
Glu Arg Leu Asn Phe Asn Leu Lys Val Asn Glu Phe Leu Lys Arg Asn
850 855 860
Lys Asp Ile Asn Ile Ile Gly Ile Asp Arg Gly Glu Arg Asn Leu Leu
865 870 875 880
Tyr Leu Val Met Ile Asn Gln Lys Gly Glu Ile Leu Lys Gln Thr Leu
885 890 895
Leu Asp Ser Met Gln Ser Gly Lys Gly Arg Pro Glu Ile Asn Tyr Lys
900 905 910
Glu Lys Leu Gln Glu Lys Glu Ile Glu Arg Asp Lys Ala Arg Lys Ser
915 920 925
Trp Gly Thr Val Glu Asn Ile Lys Glu Leu Lys Glu Gly Tyr Leu Ser
930 935 940
Ile Val Ile His Gln Ile Ser Lys Leu Met Val Glu Asn Asn Ala Ile
945 950 955 960
Val Val Leu Glu Asp Leu Asn Ile Gly Phe Lys Arg Gly Arg Gln Lys
965 970 975
Val Glu Arg Gln Val Tyr Gln Lys Phe Glu Lys Met Leu Ile Asp Lys
980 985 990
Leu Asn Phe Leu Val Phe Lys Glu Asn Lys Pro Thr Glu Pro Gly Gly
995 1000 1005
Val Leu Lys Ala Tyr Gln Leu Thr Asp Glu Phe Gln Ser Phe Glu
1010 1015 1020
Lys Leu Ser Lys Gln Thr Gly Phe Leu Phe Tyr Val Pro Ser Trp
1025 1030 1035
Asn Thr Ser Lys Ile Asp Pro Arg Thr Gly Phe Ile Asp Phe Leu
1040 1045 1050
His Pro Ala Tyr Glu Asn Ile Glu Lys Ala Lys Gln Trp Ile Asn
1055 1060 1065
Lys Phe Asp Ser Ile Arg Phe Asn Ser Lys Met Asp Trp Phe Glu
1070 1075 1080
Phe Thr Ala Asp Thr Arg Lys Phe Ser Glu Asn Leu Met Leu Gly
1085 1090 1095
Lys Asn Arg Val Trp Val Ile Cys Thr Thr Asn Val Glu Arg Tyr
1100 1105 1110
Phe Thr Ser Lys Thr Ala Asn Ser Ser Ile Gln Tyr Asn Ser Ile
1115 1120 1125
Gln Ile Thr Glu Lys Leu Lys Glu Leu Phe Val Asp Ile Pro Phe
1130 1135 1140
Ser Asn Gly Gln Asp Leu Lys Pro Glu Ile Leu Arg Lys Asn Asp
1145 1150 1155
Ala Val Phe Phe Lys Ser Leu Leu Phe Tyr Ile Lys Thr Thr Leu
1160 1165 1170
Ser Leu Arg Gln Asn Asn Gly Lys Lys Gly Glu Glu Glu Lys Asp
1175 1180 1185
Phe Ile Leu Ser Pro Val Val Asp Ser Lys Gly Arg Phe Phe Asn
1190 1195 1200
Ser Leu Glu Ala Ser Asp Asp Glu Pro Lys Asp Ala Asp Ala Asn
1205 1210 1215
Gly Ala Tyr His Ile Ala Leu Lys Gly Leu Met Asn Leu Leu Val
1220 1225 1230
Leu Asn Glu Thr Lys Glu Glu Asn Leu Ser Arg Pro Lys Trp Lys
1235 1240 1245
Ile Lys Asn Lys Asp Trp Leu Glu Phe Val Trp Glu Arg Asn Arg
1250 1255 1260
Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys
1265 1270 1275
Lys Gly Ser Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Tyr Pro Tyr
1280 1285 1290
Asp Val Pro Asp Tyr Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
1295 1300 1305
<210> 19
<211> 1272
<212> PRT
<213> 毛螺菌科细菌ND2006
<220>
<221> MISC_FEATURE
<222> (1)..(1272)
<223> LbCpf1; pY016
<400> 19
Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys Thr Leu
1 5 10 15
Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile Asp Asn
20 25 30
Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr Lys Gly
35 40 45
Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn Asp Val
50 55 60
Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile Ser Leu Phe
65 70 75 80
Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu Glu Asn Leu
85 90 95
Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys Gly Asn Glu
100 105 110
Gly Tyr Lys Ser Leu Phe Lys Lys Asp Ile Ile Glu Thr Ile Leu Pro
115 120 125
Glu Phe Leu Asp Asp Lys Asp Glu Ile Ala Leu Val Asn Ser Phe Asn
130 135 140
Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg Glu Asn Met
145 150 155 160
Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg Cys Ile Asn
165 170 175
Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe Glu Lys Val
180 185 190
Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys Glu Lys Ile
195 200 205
Leu Asn Ser Asp Tyr Asp Val Glu Asp Phe Phe Glu Gly Glu Phe Phe
210 215 220
Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn Ala Ile Ile
225 230 235 240
Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly Leu Asn Glu
245 250 255
Tyr Ile Asn Leu Tyr Asn Gln Lys Thr Lys Gln Lys Leu Pro Lys Phe
260 265 270
Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser Leu Ser Phe
275 280 285
Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu Val Phe Arg
290 295 300
Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile Lys Lys Leu
305 310 315 320
Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala Gly Ile Phe
325 330 335
Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp Ile Phe Gly
340 345 350
Glu Trp Asn Val Ile Arg Asp Lys Trp Asn Ala Glu Tyr Asp Asp Ile
355 360 365
His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu Asp Asp Arg
370 375 380
Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu Gln Leu Gln
385 390 395 400
Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu Lys Glu Ile
405 410 415
Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly Ser Ser Glu
420 425 430
Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu Lys Lys Asn
435 440 445
Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser Val Lys Ser
450 455 460
Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys Glu Thr Asn
465 470 475 480
Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr Asp Ile Leu
485 490 495
Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr Val Thr Gln
500 505 510
Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln Asn Pro Gln
515 520 525
Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr Arg Ala Thr
530 535 540
Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met Asp Lys Lys
545 550 555 560
Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val Asn Gly Asn
565 570 575
Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met Leu
580 585 590
Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr Asn Pro Ser
595 600 605
Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys Lys Gly Asp
610 615 620
Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe Phe Lys Asp
625 630 635 640
Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp Phe Asn Phe
645 650 655
Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr Arg Glu Val
660 665 670
Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser Lys Lys Glu
675 680 685
Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe Gln Ile Tyr
690 695 700
Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn Leu His Thr
705 710 715 720
Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly Gln Ile Arg
725 730 735
Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser Leu Lys Lys
740 745 750
Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala Asn Lys Asn
755 760 765
Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp Val Tyr Lys
770 775 780
Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile Pro Ile Ala
785 790 795 800
Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr Glu Val Arg
805 810 815
Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly Ile Asp Arg
820 825 830
Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly Lys Gly Asn
835 840 845
Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn Phe Asn Gly
850 855 860
Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys Lys Glu Lys
865 870 875 880
Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu Asn Ile Lys
885 890 895
Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys Ile Cys Glu
900 905 910
Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Glu Asp Leu Asn Ser
915 920 925
Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val Tyr Gln Lys
930 935 940
Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met Val Asp Lys Lys
945 950 955 960
Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys Gly Tyr Gln Ile Thr
965 970 975
Asn Lys Phe Glu Ser Phe Lys Ser Met Ser Thr Gln Asn Gly Phe Ile
980 985 990
Phe Tyr Ile Pro Ala Trp Leu Thr Ser Lys Ile Asp Pro Ser Thr Gly
995 1000 1005
Phe Val Asn Leu Leu Lys Thr Lys Tyr Thr Ser Ile Ala Asp Ser
1010 1015 1020
Lys Lys Phe Ile Ser Ser Phe Asp Arg Ile Met Tyr Val Pro Glu
1025 1030 1035
Glu Asp Leu Phe Glu Phe Ala Leu Asp Tyr Lys Asn Phe Ser Arg
1040 1045 1050
Thr Asp Ala Asp Tyr Ile Lys Lys Trp Lys Leu Tyr Ser Tyr Gly
1055 1060 1065
Asn Arg Ile Arg Ile Phe Arg Asn Pro Lys Lys Asn Asn Val Phe
1070 1075 1080
Asp Trp Glu Glu Val Cys Leu Thr Ser Ala Tyr Lys Glu Leu Phe
1085 1090 1095
Asn Lys Tyr Gly Ile Asn Tyr Gln Gln Gly Asp Ile Arg Ala Leu
1100 1105 1110
Leu Cys Glu Gln Ser Asp Lys Ala Phe Tyr Ser Ser Phe Met Ala
1115 1120 1125
Leu Met Ser Leu Met Leu Gln Met Arg Asn Ser Ile Thr Gly Arg
1130 1135 1140
Thr Asp Val Asp Phe Leu Ile Ser Pro Val Lys Asn Ser Asp Gly
1145 1150 1155
Ile Phe Tyr Asp Ser Arg Asn Tyr Glu Ala Gln Glu Asn Ala Ile
1160 1165 1170
Leu Pro Lys Asn Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala Arg
1175 1180 1185
Lys Val Leu Trp Ala Ile Gly Gln Phe Lys Lys Ala Glu Asp Glu
1190 1195 1200
Lys Leu Asp Lys Val Lys Ile Ala Ile Ser Asn Lys Glu Trp Leu
1205 1210 1215
Glu Tyr Ala Gln Thr Ser Val Lys His Lys Arg Pro Ala Ala Thr
1220 1225 1230
Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys Gly Ser Tyr Pro Tyr
1235 1240 1245
Asp Val Pro Asp Tyr Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
1250 1255 1260
Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
1265 1270
<210> 20
<211> 1305
<212> PRT
<213> 狗齿龈卟啉单胞菌
<220>
<221> MISC_FEATURE
<222> (1)..(1305)
<223> PcCpf1; pY017
<400> 20
Met Asp Ser Leu Lys Asp Phe Thr Asn Leu Tyr Pro Val Ser Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Lys Pro Val Gly Lys Thr Leu Glu Asn Ile Glu
20 25 30
Lys Ala Gly Ile Leu Lys Glu Asp Glu His Arg Ala Glu Ser Tyr Arg
35 40 45
Arg Val Lys Lys Ile Ile Asp Thr Tyr His Lys Val Phe Ile Asp Ser
50 55 60
Ser Leu Glu Asn Met Ala Lys Met Gly Ile Glu Asn Glu Ile Lys Ala
65 70 75 80
Met Leu Gln Ser Phe Cys Glu Leu Tyr Lys Lys Asp His Arg Thr Glu
85 90 95
Gly Glu Asp Lys Ala Leu Asp Lys Ile Arg Ala Val Leu Arg Gly Leu
100 105 110
Ile Val Gly Ala Phe Thr Gly Val Cys Gly Arg Arg Glu Asn Thr Val
115 120 125
Gln Asn Glu Lys Tyr Glu Ser Leu Phe Lys Glu Lys Leu Ile Lys Glu
130 135 140
Ile Leu Pro Asp Phe Val Leu Ser Thr Glu Ala Glu Ser Leu Pro Phe
145 150 155 160
Ser Val Glu Glu Ala Thr Arg Ser Leu Lys Glu Phe Asp Ser Phe Thr
165 170 175
Ser Tyr Phe Ala Gly Phe Tyr Glu Asn Arg Lys Asn Ile Tyr Ser Thr
180 185 190
Lys Pro Gln Ser Thr Ala Ile Ala Tyr Arg Leu Ile His Glu Asn Leu
195 200 205
Pro Lys Phe Ile Asp Asn Ile Leu Val Phe Gln Lys Ile Lys Glu Pro
210 215 220
Ile Ala Lys Glu Leu Glu His Ile Arg Ala Asp Phe Ser Ala Gly Gly
225 230 235 240
Tyr Ile Lys Lys Asp Glu Arg Leu Glu Asp Ile Phe Ser Leu Asn Tyr
245 250 255
Tyr Ile His Val Leu Ser Gln Ala Gly Ile Glu Lys Tyr Asn Ala Leu
260 265 270
Ile Gly Lys Ile Val Thr Glu Gly Asp Gly Glu Met Lys Gly Leu Asn
275 280 285
Glu His Ile Asn Leu Tyr Asn Gln Gln Arg Gly Arg Glu Asp Arg Leu
290 295 300
Pro Leu Phe Arg Pro Leu Tyr Lys Gln Ile Leu Ser Asp Arg Glu Gln
305 310 315 320
Leu Ser Tyr Leu Pro Glu Ser Phe Glu Lys Asp Glu Glu Leu Leu Arg
325 330 335
Ala Leu Lys Glu Phe Tyr Asp His Ile Ala Glu Asp Ile Leu Gly Arg
340 345 350
Thr Gln Gln Leu Met Thr Ser Ile Ser Glu Tyr Asp Leu Ser Arg Ile
355 360 365
Tyr Val Arg Asn Asp Ser Gln Leu Thr Asp Ile Ser Lys Lys Met Leu
370 375 380
Gly Asp Trp Asn Ala Ile Tyr Met Ala Arg Glu Arg Ala Tyr Asp His
385 390 395 400
Glu Gln Ala Pro Lys Arg Ile Thr Ala Lys Tyr Glu Arg Asp Arg Ile
405 410 415
Lys Ala Leu Lys Gly Glu Glu Ser Ile Ser Leu Ala Asn Leu Asn Ser
420 425 430
Cys Ile Ala Phe Leu Asp Asn Val Arg Asp Cys Arg Val Asp Thr Tyr
435 440 445
Leu Ser Thr Leu Gly Gln Lys Glu Gly Pro His Gly Leu Ser Asn Leu
450 455 460
Val Glu Asn Val Phe Ala Ser Tyr His Glu Ala Glu Gln Leu Leu Ser
465 470 475 480
Phe Pro Tyr Pro Glu Glu Asn Asn Leu Ile Gln Asp Lys Asp Asn Val
485 490 495
Val Leu Ile Lys Asn Leu Leu Asp Asn Ile Ser Asp Leu Gln Arg Phe
500 505 510
Leu Lys Pro Leu Trp Gly Met Gly Asp Glu Pro Asp Lys Asp Glu Arg
515 520 525
Phe Tyr Gly Glu Tyr Asn Tyr Ile Arg Gly Ala Leu Asp Gln Val Ile
530 535 540
Pro Leu Tyr Asn Lys Val Arg Asn Tyr Leu Thr Arg Lys Pro Tyr Ser
545 550 555 560
Thr Arg Lys Val Lys Leu Asn Phe Gly Asn Ser Gln Leu Leu Ser Gly
565 570 575
Trp Asp Arg Asn Lys Glu Lys Asp Asn Ser Cys Val Ile Leu Arg Lys
580 585 590
Gly Gln Asn Phe Tyr Leu Ala Ile Met Asn Asn Arg His Lys Arg Ser
595 600 605
Phe Glu Asn Lys Met Leu Pro Glu Tyr Lys Glu Gly Glu Pro Tyr Phe
610 615 620
Glu Lys Met Asp Tyr Lys Phe Leu Pro Asp Pro Asn Lys Met Leu Pro
625 630 635 640
Lys Val Phe Leu Ser Lys Lys Gly Ile Glu Ile Tyr Lys Pro Ser Pro
645 650 655
Lys Leu Leu Glu Gln Tyr Gly His Gly Thr His Lys Lys Gly Asp Thr
660 665 670
Phe Ser Met Asp Asp Leu His Glu Leu Ile Asp Phe Phe Lys His Ser
675 680 685
Ile Glu Ala His Glu Asp Trp Lys Gln Phe Gly Phe Lys Phe Ser Asp
690 695 700
Thr Ala Thr Tyr Glu Asn Val Ser Ser Phe Tyr Arg Glu Val Glu Asp
705 710 715 720
Gln Gly Tyr Lys Leu Ser Phe Arg Lys Val Ser Glu Ser Tyr Val Tyr
725 730 735
Ser Leu Ile Asp Gln Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys
740 745 750
Asp Phe Ser Pro Cys Ser Lys Gly Thr Pro Asn Leu His Thr Leu Tyr
755 760 765
Trp Arg Met Leu Phe Asp Glu Arg Asn Leu Ala Asp Val Ile Tyr Lys
770 775 780
Leu Asp Gly Lys Ala Glu Ile Phe Phe Arg Glu Lys Ser Leu Lys Asn
785 790 795 800
Asp His Pro Thr His Pro Ala Gly Lys Pro Ile Lys Lys Lys Ser Arg
805 810 815
Gln Lys Lys Gly Glu Glu Ser Leu Phe Glu Tyr Asp Leu Val Lys Asp
820 825 830
Arg Arg Tyr Thr Met Asp Lys Phe Gln Phe His Val Pro Ile Thr Met
835 840 845
Asn Phe Lys Cys Ser Ala Gly Ser Lys Val Asn Asp Met Val Asn Ala
850 855 860
His Ile Arg Glu Ala Lys Asp Met His Val Ile Gly Ile Asp Arg Gly
865 870 875 880
Glu Arg Asn Leu Leu Tyr Ile Cys Val Ile Asp Ser Arg Gly Thr Ile
885 890 895
Leu Asp Gln Ile Ser Leu Asn Thr Ile Asn Asp Ile Asp Tyr His Asp
900 905 910
Leu Leu Glu Ser Arg Asp Lys Asp Arg Gln Gln Glu His Arg Asn Trp
915 920 925
Gln Thr Ile Glu Gly Ile Lys Glu Leu Lys Gln Gly Tyr Leu Ser Gln
930 935 940
Ala Val His Arg Ile Ala Glu Leu Met Val Ala Tyr Lys Ala Val Val
945 950 955 960
Ala Leu Glu Asp Leu Asn Met Gly Phe Lys Arg Gly Arg Gln Lys Val
965 970 975
Glu Ser Ser Val Tyr Gln Gln Phe Glu Lys Gln Leu Ile Asp Lys Leu
980 985 990
Asn Tyr Leu Val Asp Lys Lys Lys Arg Pro Glu Asp Ile Gly Gly Leu
995 1000 1005
Leu Arg Ala Tyr Gln Phe Thr Ala Pro Phe Lys Ser Phe Lys Glu
1010 1015 1020
Met Gly Lys Gln Asn Gly Phe Leu Phe Tyr Ile Pro Ala Trp Asn
1025 1030 1035
Thr Ser Asn Ile Asp Pro Thr Thr Gly Phe Val Asn Leu Phe His
1040 1045 1050
Val Gln Tyr Glu Asn Val Asp Lys Ala Lys Ser Phe Phe Gln Lys
1055 1060 1065
Phe Asp Ser Ile Ser Tyr Asn Pro Lys Lys Asp Trp Phe Glu Phe
1070 1075 1080
Ala Phe Asp Tyr Lys Asn Phe Thr Lys Lys Ala Glu Gly Ser Arg
1085 1090 1095
Ser Met Trp Ile Leu Cys Thr His Gly Ser Arg Ile Lys Asn Phe
1100 1105 1110
Arg Asn Ser Gln Lys Asn Gly Gln Trp Asp Ser Glu Glu Phe Ala
1115 1120 1125
Leu Thr Glu Ala Phe Lys Ser Leu Phe Val Arg Tyr Glu Ile Asp
1130 1135 1140
Tyr Thr Ala Asp Leu Lys Thr Ala Ile Val Asp Glu Lys Gln Lys
1145 1150 1155
Asp Phe Phe Val Asp Leu Leu Lys Leu Phe Lys Leu Thr Val Gln
1160 1165 1170
Met Arg Asn Ser Trp Lys Glu Lys Asp Leu Asp Tyr Leu Ile Ser
1175 1180 1185
Pro Val Ala Gly Ala Asp Gly Arg Phe Phe Asp Thr Arg Glu Gly
1190 1195 1200
Asn Lys Ser Leu Pro Lys Asp Ala Asp Ala Asn Gly Ala Tyr Asn
1205 1210 1215
Ile Ala Leu Lys Gly Leu Trp Ala Leu Arg Gln Ile Arg Gln Thr
1220 1225 1230
Ser Glu Gly Gly Lys Leu Lys Leu Ala Ile Ser Asn Lys Glu Trp
1235 1240 1245
Leu Gln Phe Val Gln Glu Arg Ser Tyr Glu Lys Asp Lys Arg Pro
1250 1255 1260
Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys Gly Ser
1265 1270 1275
Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Tyr Pro Tyr Asp Val Pro
1280 1285 1290
Asp Tyr Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
1295 1300 1305
<210> 21
<211> 1368
<212> PRT
<213> 解糖胨普雷沃氏菌
<220>
<221> MISC_FEATURE
<222> (1)..(1368)
<223> PdCpf1; pY018
<400> 21
Met Glu Asn Tyr Gln Glu Phe Thr Asn Leu Phe Gln Leu Asn Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Lys Pro Ile Gly Lys Thr Cys Glu Leu Leu Glu
20 25 30
Glu Gly Lys Ile Phe Ala Ser Gly Ser Phe Leu Glu Lys Asp Lys Val
35 40 45
Arg Ala Asp Asn Val Ser Tyr Val Lys Lys Glu Ile Asp Lys Lys His
50 55 60
Lys Ile Phe Ile Glu Glu Thr Leu Ser Ser Phe Ser Ile Ser Asn Asp
65 70 75 80
Leu Leu Lys Gln Tyr Phe Asp Cys Tyr Asn Glu Leu Lys Ala Phe Lys
85 90 95
Lys Asp Cys Lys Ser Asp Glu Glu Glu Val Lys Lys Thr Ala Leu Arg
100 105 110
Asn Lys Cys Thr Ser Ile Gln Arg Ala Met Arg Glu Ala Ile Ser Gln
115 120 125
Ala Phe Leu Lys Ser Pro Gln Lys Lys Leu Leu Ala Ile Lys Asn Leu
130 135 140
Ile Glu Asn Val Phe Lys Ala Asp Glu Asn Val Gln His Phe Ser Glu
145 150 155 160
Phe Thr Ser Tyr Phe Ser Gly Phe Glu Thr Asn Arg Glu Asn Phe Tyr
165 170 175
Ser Asp Glu Glu Lys Ser Thr Ser Ile Ala Tyr Arg Leu Val His Asp
180 185 190
Asn Leu Pro Ile Phe Ile Lys Asn Ile Tyr Ile Phe Glu Lys Leu Lys
195 200 205
Glu Gln Phe Asp Ala Lys Thr Leu Ser Glu Ile Phe Glu Asn Tyr Lys
210 215 220
Leu Tyr Val Ala Gly Ser Ser Leu Asp Glu Val Phe Ser Leu Glu Tyr
225 230 235 240
Phe Asn Asn Thr Leu Thr Gln Lys Gly Ile Asp Asn Tyr Asn Ala Val
245 250 255
Ile Gly Lys Ile Val Lys Glu Asp Lys Gln Glu Ile Gln Gly Leu Asn
260 265 270
Glu His Ile Asn Leu Tyr Asn Gln Lys His Lys Asp Arg Arg Leu Pro
275 280 285
Phe Phe Ile Ser Leu Lys Lys Gln Ile Leu Ser Asp Arg Glu Ala Leu
290 295 300
Ser Trp Leu Pro Asp Met Phe Lys Asn Asp Ser Glu Val Ile Lys Ala
305 310 315 320
Leu Lys Gly Phe Tyr Ile Glu Asp Gly Phe Glu Asn Asn Val Leu Thr
325 330 335
Pro Leu Ala Thr Leu Leu Ser Ser Leu Asp Lys Tyr Asn Leu Asn Gly
340 345 350
Ile Phe Ile Arg Asn Asn Glu Ala Leu Ser Ser Leu Ser Gln Asn Val
355 360 365
Tyr Arg Asn Phe Ser Ile Asp Glu Ala Ile Asp Ala Asn Ala Glu Leu
370 375 380
Gln Thr Phe Asn Asn Tyr Glu Leu Ile Ala Asn Ala Leu Arg Ala Lys
385 390 395 400
Ile Lys Lys Glu Thr Lys Gln Gly Arg Lys Ser Phe Glu Lys Tyr Glu
405 410 415
Glu Tyr Ile Asp Lys Lys Val Lys Ala Ile Asp Ser Leu Ser Ile Gln
420 425 430
Glu Ile Asn Glu Leu Val Glu Asn Tyr Val Ser Glu Phe Asn Ser Asn
435 440 445
Ser Gly Asn Met Pro Arg Lys Val Glu Asp Tyr Phe Ser Leu Met Arg
450 455 460
Lys Gly Asp Phe Gly Ser Asn Asp Leu Ile Glu Asn Ile Lys Thr Lys
465 470 475 480
Leu Ser Ala Ala Glu Lys Leu Leu Gly Thr Lys Tyr Gln Glu Thr Ala
485 490 495
Lys Asp Ile Phe Lys Lys Asp Glu Asn Ser Lys Leu Ile Lys Glu Leu
500 505 510
Leu Asp Ala Thr Lys Gln Phe Gln His Phe Ile Lys Pro Leu Leu Gly
515 520 525
Thr Gly Glu Glu Ala Asp Arg Asp Leu Val Phe Tyr Gly Asp Phe Leu
530 535 540
Pro Leu Tyr Glu Lys Phe Glu Glu Leu Thr Leu Leu Tyr Asn Lys Val
545 550 555 560
Arg Asn Arg Leu Thr Gln Lys Pro Tyr Ser Lys Asp Lys Ile Arg Leu
565 570 575
Cys Phe Asn Lys Pro Lys Leu Met Thr Gly Trp Val Asp Ser Lys Thr
580 585 590
Glu Lys Ser Asp Asn Gly Thr Gln Tyr Gly Gly Tyr Leu Phe Arg Lys
595 600 605
Lys Asn Glu Ile Gly Glu Tyr Asp Tyr Phe Leu Gly Ile Ser Ser Lys
610 615 620
Ala Gln Leu Phe Arg Lys Asn Glu Ala Val Ile Gly Asp Tyr Glu Arg
625 630 635 640
Leu Asp Tyr Tyr Gln Pro Lys Ala Asn Thr Ile Tyr Gly Ser Ala Tyr
645 650 655
Glu Gly Glu Asn Ser Tyr Lys Glu Asp Lys Lys Arg Leu Asn Lys Val
660 665 670
Ile Ile Ala Tyr Ile Glu Gln Ile Lys Gln Thr Asn Ile Lys Lys Ser
675 680 685
Ile Ile Glu Ser Ile Ser Lys Tyr Pro Asn Ile Ser Asp Asp Asp Lys
690 695 700
Val Thr Pro Ser Ser Leu Leu Glu Lys Ile Lys Lys Val Ser Ile Asp
705 710 715 720
Ser Tyr Asn Gly Ile Leu Ser Phe Lys Ser Phe Gln Ser Val Asn Lys
725 730 735
Glu Val Ile Asp Asn Leu Leu Lys Thr Ile Ser Pro Leu Lys Asn Lys
740 745 750
Ala Glu Phe Leu Asp Leu Ile Asn Lys Asp Tyr Gln Ile Phe Thr Glu
755 760 765
Val Gln Ala Val Ile Asp Glu Ile Cys Lys Gln Lys Thr Phe Ile Tyr
770 775 780
Phe Pro Ile Ser Asn Val Glu Leu Glu Lys Glu Met Gly Asp Lys Asp
785 790 795 800
Lys Pro Leu Cys Leu Phe Gln Ile Ser Asn Lys Asp Leu Ser Phe Ala
805 810 815
Lys Thr Phe Ser Ala Asn Leu Arg Lys Lys Arg Gly Ala Glu Asn Leu
820 825 830
His Thr Met Leu Phe Lys Ala Leu Met Glu Gly Asn Gln Asp Asn Leu
835 840 845
Asp Leu Gly Ser Gly Ala Ile Phe Tyr Arg Ala Lys Ser Leu Asp Gly
850 855 860
Asn Lys Pro Thr His Pro Ala Asn Glu Ala Ile Lys Cys Arg Asn Val
865 870 875 880
Ala Asn Lys Asp Lys Val Ser Leu Phe Thr Tyr Asp Ile Tyr Lys Asn
885 890 895
Arg Arg Tyr Met Glu Asn Lys Phe Leu Phe His Leu Ser Ile Val Gln
900 905 910
Asn Tyr Lys Ala Ala Asn Asp Ser Ala Gln Leu Asn Ser Ser Ala Thr
915 920 925
Glu Tyr Ile Arg Lys Ala Asp Asp Leu His Ile Ile Gly Ile Asp Arg
930 935 940
Gly Glu Arg Asn Leu Leu Tyr Tyr Ser Val Ile Asp Met Lys Gly Asn
945 950 955 960
Ile Val Glu Gln Asp Ser Leu Asn Ile Ile Arg Asn Asn Asp Leu Glu
965 970 975
Thr Asp Tyr His Asp Leu Leu Asp Lys Arg Glu Lys Glu Arg Lys Ala
980 985 990
Asn Arg Gln Asn Trp Glu Ala Val Glu Gly Ile Lys Asp Leu Lys Lys
995 1000 1005
Gly Tyr Leu Ser Gln Ala Val His Gln Ile Ala Gln Leu Met Leu
1010 1015 1020
Lys Tyr Asn Ala Ile Ile Ala Leu Glu Asp Leu Gly Gln Met Phe
1025 1030 1035
Val Thr Arg Gly Gln Lys Ile Glu Lys Ala Val Tyr Gln Gln Phe
1040 1045 1050
Glu Lys Ser Leu Val Asp Lys Leu Ser Tyr Leu Val Asp Lys Lys
1055 1060 1065
Arg Pro Tyr Asn Glu Leu Gly Gly Ile Leu Lys Ala Tyr Gln Leu
1070 1075 1080
Ala Ser Ser Ile Thr Lys Asn Asn Ser Asp Lys Gln Asn Gly Phe
1085 1090 1095
Leu Phe Tyr Val Pro Ala Trp Asn Thr Ser Lys Ile Asp Pro Val
1100 1105 1110
Thr Gly Phe Thr Asp Leu Leu Arg Pro Lys Ala Met Thr Ile Lys
1115 1120 1125
Glu Ala Gln Asp Phe Phe Gly Ala Phe Asp Asn Ile Ser Tyr Asn
1130 1135 1140
Asp Lys Gly Tyr Phe Glu Phe Glu Thr Asn Tyr Asp Lys Phe Lys
1145 1150 1155
Ile Arg Met Lys Ser Ala Gln Thr Arg Trp Thr Ile Cys Thr Phe
1160 1165 1170
Gly Asn Arg Ile Lys Arg Lys Lys Asp Lys Asn Tyr Trp Asn Tyr
1175 1180 1185
Glu Glu Val Glu Leu Thr Glu Glu Phe Lys Lys Leu Phe Lys Asp
1190 1195 1200
Ser Asn Ile Asp Tyr Glu Asn Cys Asn Leu Lys Glu Glu Ile Gln
1205 1210 1215
Asn Lys Asp Asn Arg Lys Phe Phe Asp Asp Leu Ile Lys Leu Leu
1220 1225 1230
Gln Leu Thr Leu Gln Met Arg Asn Ser Asp Asp Lys Gly Asn Asp
1235 1240 1245
Tyr Ile Ile Ser Pro Val Ala Asn Ala Glu Gly Gln Phe Phe Asp
1250 1255 1260
Ser Arg Asn Gly Asp Lys Lys Leu Pro Leu Asp Ala Asp Ala Asn
1265 1270 1275
Gly Ala Tyr Asn Ile Ala Arg Lys Gly Leu Trp Asn Ile Arg Gln
1280 1285 1290
Ile Lys Gln Thr Lys Asn Asp Lys Lys Leu Asn Leu Ser Ile Ser
1295 1300 1305
Ser Thr Glu Trp Leu Asp Phe Val Arg Glu Lys Pro Tyr Leu Lys
1310 1315 1320
Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys
1325 1330 1335
Lys Gly Ser Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Tyr Pro Tyr
1340 1345 1350
Asp Val Pro Asp Tyr Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
1355 1360 1365
<210> 22
<211> 1291
<212> PRT
<213> 猕猴卟啉单胞菌
<220>
<221> MISC_FEATURE
<222> (1)..(1291)
<223> PmCpf1; pY09
<400> 22
Met Lys Thr Gln His Phe Phe Glu Asp Phe Thr Ser Leu Tyr Ser Leu
1 5 10 15
Ser Lys Thr Ile Arg Phe Glu Leu Lys Pro Ile Gly Lys Thr Leu Glu
20 25 30
Asn Ile Lys Lys Asn Gly Leu Ile Arg Arg Asp Glu Gln Arg Leu Asp
35 40 45
Asp Tyr Glu Lys Leu Lys Lys Val Ile Asp Glu Tyr His Glu Asp Phe
50 55 60
Ile Ala Asn Ile Leu Ser Ser Phe Ser Phe Ser Glu Glu Ile Leu Gln
65 70 75 80
Ser Tyr Ile Gln Asn Leu Ser Glu Ser Glu Ala Arg Ala Lys Ile Glu
85 90 95
Lys Thr Met Arg Asp Thr Leu Ala Lys Ala Phe Ser Glu Asp Glu Arg
100 105 110
Tyr Lys Ser Ile Phe Lys Lys Glu Leu Val Lys Lys Asp Ile Pro Val
115 120 125
Trp Cys Pro Ala Tyr Lys Ser Leu Cys Lys Lys Phe Asp Asn Phe Thr
130 135 140
Thr Ser Leu Val Pro Phe His Glu Asn Arg Lys Asn Leu Tyr Thr Ser
145 150 155 160
Asn Glu Ile Thr Ala Ser Ile Pro Tyr Arg Ile Val His Val Asn Leu
165 170 175
Pro Lys Phe Ile Gln Asn Ile Glu Ala Leu Cys Glu Leu Gln Lys Lys
180 185 190
Met Gly Ala Asp Leu Tyr Leu Glu Met Met Glu Asn Leu Arg Asn Val
195 200 205
Trp Pro Ser Phe Val Lys Thr Pro Asp Asp Leu Cys Asn Leu Lys Thr
210 215 220
Tyr Asn His Leu Met Val Gln Ser Ser Ile Ser Glu Tyr Asn Arg Phe
225 230 235 240
Val Gly Gly Tyr Ser Thr Glu Asp Gly Thr Lys His Gln Gly Ile Asn
245 250 255
Glu Trp Ile Asn Ile Tyr Arg Gln Arg Asn Lys Glu Met Arg Leu Pro
260 265 270
Gly Leu Val Phe Leu His Lys Gln Ile Leu Ala Lys Val Asp Ser Ser
275 280 285
Ser Phe Ile Ser Asp Thr Leu Glu Asn Asp Asp Gln Val Phe Cys Val
290 295 300
Leu Arg Gln Phe Arg Lys Leu Phe Trp Asn Thr Val Ser Ser Lys Glu
305 310 315 320
Asp Asp Ala Ala Ser Leu Lys Asp Leu Phe Cys Gly Leu Ser Gly Tyr
325 330 335
Asp Pro Glu Ala Ile Tyr Val Ser Asp Ala His Leu Ala Thr Ile Ser
340 345 350
Lys Asn Ile Phe Asp Arg Trp Asn Tyr Ile Ser Asp Ala Ile Arg Arg
355 360 365
Lys Thr Glu Val Leu Met Pro Arg Lys Lys Glu Ser Val Glu Arg Tyr
370 375 380
Ala Glu Lys Ile Ser Lys Gln Ile Lys Lys Arg Gln Ser Tyr Ser Leu
385 390 395 400
Ala Glu Leu Asp Asp Leu Leu Ala His Tyr Ser Glu Glu Ser Leu Pro
405 410 415
Ala Gly Phe Ser Leu Leu Ser Tyr Phe Thr Ser Leu Gly Gly Gln Lys
420 425 430
Tyr Leu Val Ser Asp Gly Glu Val Ile Leu Tyr Glu Glu Gly Ser Asn
435 440 445
Ile Trp Asp Glu Val Leu Ile Ala Phe Arg Asp Leu Gln Val Ile Leu
450 455 460
Asp Lys Asp Phe Thr Glu Lys Lys Leu Gly Lys Asp Glu Glu Ala Val
465 470 475 480
Ser Val Ile Lys Lys Ala Leu Asp Ser Ala Leu Arg Leu Arg Lys Phe
485 490 495
Phe Asp Leu Leu Ser Gly Thr Gly Ala Glu Ile Arg Arg Asp Ser Ser
500 505 510
Phe Tyr Ala Leu Tyr Thr Asp Arg Met Asp Lys Leu Lys Gly Leu Leu
515 520 525
Lys Met Tyr Asp Lys Val Arg Asn Tyr Leu Thr Lys Lys Pro Tyr Ser
530 535 540
Ile Glu Lys Phe Lys Leu His Phe Asp Asn Pro Ser Leu Leu Ser Gly
545 550 555 560
Trp Asp Lys Asn Lys Glu Leu Asn Asn Leu Ser Val Ile Phe Arg Gln
565 570 575
Asn Gly Tyr Tyr Tyr Leu Gly Ile Met Thr Pro Lys Gly Lys Asn Leu
580 585 590
Phe Lys Thr Leu Pro Lys Leu Gly Ala Glu Glu Met Phe Tyr Glu Lys
595 600 605
Met Glu Tyr Lys Gln Ile Ala Glu Pro Met Leu Met Leu Pro Lys Val
610 615 620
Phe Phe Pro Lys Lys Thr Lys Pro Ala Phe Ala Pro Asp Gln Ser Val
625 630 635 640
Val Asp Ile Tyr Asn Lys Lys Thr Phe Lys Thr Gly Gln Lys Gly Phe
645 650 655
Asn Lys Lys Asp Leu Tyr Arg Leu Ile Asp Phe Tyr Lys Glu Ala Leu
660 665 670
Thr Val His Glu Trp Lys Leu Phe Asn Phe Ser Phe Ser Pro Thr Glu
675 680 685
Gln Tyr Arg Asn Ile Gly Glu Phe Phe Asp Glu Val Arg Glu Gln Ala
690 695 700
Tyr Lys Val Ser Met Val Asn Val Pro Ala Ser Tyr Ile Asp Glu Ala
705 710 715 720
Val Glu Asn Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe
725 730 735
Ser Pro Tyr Ser Lys Gly Ile Pro Asn Leu His Thr Leu Tyr Trp Lys
740 745 750
Ala Leu Phe Ser Glu Gln Asn Gln Ser Arg Val Tyr Lys Leu Cys Gly
755 760 765
Gly Gly Glu Leu Phe Tyr Arg Lys Ala Ser Leu His Met Gln Asp Thr
770 775 780
Thr Val His Pro Lys Gly Ile Ser Ile His Lys Lys Asn Leu Asn Lys
785 790 795 800
Lys Gly Glu Thr Ser Leu Phe Asn Tyr Asp Leu Val Lys Asp Lys Arg
805 810 815
Phe Thr Glu Asp Lys Phe Phe Phe His Val Pro Ile Ser Ile Asn Tyr
820 825 830
Lys Asn Lys Lys Ile Thr Asn Val Asn Gln Met Val Arg Asp Tyr Ile
835 840 845
Ala Gln Asn Asp Asp Leu Gln Ile Ile Gly Ile Asp Arg Gly Glu Arg
850 855 860
Asn Leu Leu Tyr Ile Ser Arg Ile Asp Thr Arg Gly Asn Leu Leu Glu
865 870 875 880
Gln Phe Ser Leu Asn Val Ile Glu Ser Asp Lys Gly Asp Leu Arg Thr
885 890 895
Asp Tyr Gln Lys Ile Leu Gly Asp Arg Glu Gln Glu Arg Leu Arg Arg
900 905 910
Arg Gln Glu Trp Lys Ser Ile Glu Ser Ile Lys Asp Leu Lys Asp Gly
915 920 925
Tyr Met Ser Gln Val Val His Lys Ile Cys Asn Met Val Val Glu His
930 935 940
Lys Ala Ile Val Val Leu Glu Asn Leu Asn Leu Ser Phe Met Lys Gly
945 950 955 960
Arg Lys Lys Val Glu Lys Ser Val Tyr Glu Lys Phe Glu Arg Met Leu
965 970 975
Val Asp Lys Leu Asn Tyr Leu Val Val Asp Lys Lys Asn Leu Ser Asn
980 985 990
Glu Pro Gly Gly Leu Tyr Ala Ala Tyr Gln Leu Thr Asn Pro Leu Phe
995 1000 1005
Ser Phe Glu Glu Leu His Arg Tyr Pro Gln Ser Gly Ile Leu Phe
1010 1015 1020
Phe Val Asp Pro Trp Asn Thr Ser Leu Thr Asp Pro Ser Thr Gly
1025 1030 1035
Phe Val Asn Leu Leu Gly Arg Ile Asn Tyr Thr Asn Val Gly Asp
1040 1045 1050
Ala Arg Lys Phe Phe Asp Arg Phe Asn Ala Ile Arg Tyr Asp Gly
1055 1060 1065
Lys Gly Asn Ile Leu Phe Asp Leu Asp Leu Ser Arg Phe Asp Val
1070 1075 1080
Arg Val Glu Thr Gln Arg Lys Leu Trp Thr Leu Thr Thr Phe Gly
1085 1090 1095
Ser Arg Ile Ala Lys Ser Lys Lys Ser Gly Lys Trp Met Val Glu
1100 1105 1110
Arg Ile Glu Asn Leu Ser Leu Cys Phe Leu Glu Leu Phe Glu Gln
1115 1120 1125
Phe Asn Ile Gly Tyr Arg Val Glu Lys Asp Leu Lys Lys Ala Ile
1130 1135 1140
Leu Ser Gln Asp Arg Lys Glu Phe Tyr Val Arg Leu Ile Tyr Leu
1145 1150 1155
Phe Asn Leu Met Met Gln Ile Arg Asn Ser Asp Gly Glu Glu Asp
1160 1165 1170
Tyr Ile Leu Ser Pro Ala Leu Asn Glu Lys Asn Leu Gln Phe Asp
1175 1180 1185
Ser Arg Leu Ile Glu Ala Lys Asp Leu Pro Val Asp Ala Asp Ala
1190 1195 1200
Asn Gly Ala Tyr Asn Val Ala Arg Lys Gly Leu Met Val Val Gln
1205 1210 1215
Arg Ile Lys Arg Gly Asp His Glu Ser Ile His Arg Ile Gly Arg
1220 1225 1230
Ala Gln Trp Leu Arg Tyr Val Gln Glu Gly Ile Val Glu Lys Arg
1235 1240 1245
Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys Gly
1250 1255 1260
Ser Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Tyr Pro Tyr Asp Val
1265 1270 1275
Pro Asp Tyr Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
1280 1285 1290
<210> 23
<211> 399
<212> PRT
<213> 蜃楼弗朗西斯菌亚种
<220>
<221> MISC_FEATURE
<222> (1)..(399)
<223> Genbank ABZ87876 Cpf1
<400> 23
Met Phe Tyr Leu Lys Gln Ile Leu Ser Asp Thr Glu Ser Lys Ser Phe
1 5 10 15
Val Ile Asp Lys Leu Glu Asp Asp Ser Asp Val Val Thr Thr Ile Gln
20 25 30
Ser Phe Tyr Glu Gln Ile Ala Ala Phe Lys Thr Leu Glu Glu Lys Ser
35 40 45
Ile Lys Glu Thr Leu Ser Leu Leu Phe Asp Asp Leu Lys Ala Gln Lys
50 55 60
Leu Asp Leu Ser Lys Ile Tyr Phe Lys Asn Asp Lys Ser Leu Thr Asp
65 70 75 80
Leu Ser Gln Gln Val Phe Asp Asp Tyr Ser Leu Ile Gly Thr Ser Val
85 90 95
Leu Glu Tyr Ile Thr Gln Gln Val Ala Pro Lys Asn Leu Asp Asn Pro
100 105 110
Ser Lys Lys Glu Gln Glu Leu Ile Ala Lys Lys Thr Glu Lys Ala Lys
115 120 125
Tyr Leu Ser Leu Glu Thr Ile Arg Asp Ala Leu Asn Glu Phe Asn Lys
130 135 140
His Arg Asp Ile Asp Lys Gln Cys Arg Phe Glu Glu Ile Phe Ala Ser
145 150 155 160
Phe Ala Asp Ile Pro Val Leu Phe Asp Glu Ile Ala Gln Asn Lys Asn
165 170 175
Asn Leu Ala Gln Ile Ser Ile Lys Tyr Gln Asn Gln Gly Lys Lys Asp
180 185 190
Leu Leu Gln Thr Ser Ala Glu Val Asp Val Lys Ala Ile Lys Asp Leu
195 200 205
Leu Asp Gln Thr Asn Asn Leu Leu His Lys Leu Lys Ile Phe His Ile
210 215 220
Thr Gln Ser Glu Asp Lys Ala Asn Ile Leu Asp Lys Asp Glu His Phe
225 230 235 240
Tyr Leu Val Phe Asp Glu Cys Tyr Phe Glu Leu Ala Asn Ile Val Pro
245 250 255
Leu Tyr Asn Lys Ile Arg Asn Tyr Ile Thr Gln Lys Pro Tyr Ser Asp
260 265 270
Glu Lys Phe Lys Leu Asn Phe Glu Asn Ser Thr Leu Ala Asn Gly Trp
275 280 285
Asp Lys Asn Lys Glu Pro Asp Asn Thr Ala Ile Leu Phe Ile Lys Asp
290 295 300
Asp Lys Tyr Tyr Leu Gly Val Met Asn Lys Lys Asn Asn Lys Ile Phe
305 310 315 320
Asp Asp Lys Ala Ile Lys Glu Asn Lys Gly Glu Gly Tyr Lys Lys Val
325 330 335
Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro Lys Val Phe
340 345 350
Phe Ser Ala Lys Ser Ile Asn Phe Tyr Asn Pro Ser Glu Asp Ile Leu
355 360 365
Arg Ile Arg Asn His Ser Thr His Thr Lys Asn Gly Ser Pro Gln Lys
370 375 380
Asp Met Lys Asn Leu Ser Leu Ile Leu Lys Ile Ala Glu Asn Leu
385 390 395
<210> 24
<211> 485
<212> PRT
<213> 蜃楼弗朗西斯菌亚种
<220>
<221> MISC_FEATURE
<222> (1)..(485)
<223> Genbank ABZ87877 Cpf1
<400> 24
Met Val Tyr Lys Leu Asn Gly Glu Ala Glu Leu Phe Tyr Arg Lys Gln
1 5 10 15
Ser Ile Pro Lys Lys Ile Thr His Pro Ala Lys Gln Ala Ile Ala Asn
20 25 30
Lys Asn Lys Asp Asn Pro Lys Lys Glu Ser Phe Phe Glu Tyr Asp Leu
35 40 45
Ile Lys Asp Lys Arg Phe Thr Glu Asp Lys Phe Phe Phe His Cys Pro
50 55 60
Ile Thr Met Asn Phe Lys Ser Ser Gly Ala Asn Lys Phe Asn Asp Glu
65 70 75 80
Val Asn Leu Leu Leu Lys Glu Lys Ala Asn Asp Val His Ile Leu Ser
85 90 95
Ile Asp Arg Gly Glu Arg His Leu Ala Tyr Tyr Thr Leu Val Asp Ser
100 105 110
Lys Gly Asn Ile Ile Lys Gln Asp Thr Phe Asn Ile Ile Gly Asn Asp
115 120 125
Arg Met Lys Thr Asn Tyr His Asp Lys Leu Ala Ala Ile Glu Lys Asp
130 135 140
Arg Glu Ser Ala Arg Lys Asp Trp Lys Lys Ile Asn Asn Ile Lys Glu
145 150 155 160
Met Lys Glu Gly Tyr Leu Ser Gln Val Val His Glu Ile Ala Lys Leu
165 170 175
Val Ile Glu Tyr Asn Ala Ile Val Val Phe Glu Asp Leu Asn Phe Gly
180 185 190
Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Val Tyr Gln Lys Leu
195 200 205
Glu Lys Met Leu Ile Glu Lys Leu Asn Tyr Leu Val Phe Lys Asp Asn
210 215 220
Glu Phe Asp Lys Ala Gly Gly Val Leu Arg Ala Tyr Gln Leu Thr Ala
225 230 235 240
Pro Phe Glu Thr Phe Lys Lys Met Gly Lys Gln Thr Gly Val Ile Tyr
245 250 255
Tyr Val Pro Ala Gly Phe Thr Ser Lys Ile Cys Pro Val Thr Gly Phe
260 265 270
Val Asn Gln Leu Tyr Pro Lys Tyr Glu Ser Val Ser Lys Ser Gln Glu
275 280 285
Phe Phe Ser Lys Phe Asp Lys Ile Cys Tyr Asn Leu Asp Lys Gly Tyr
290 295 300
Phe Glu Phe Ser Phe Asp Tyr Lys Asn Phe Gly Asp Lys Ala Ala Lys
305 310 315 320
Gly Lys Trp Thr Ile Ala Ser Phe Gly Ser Arg Leu Ile Asn Phe Arg
325 330 335
Asn Ser Asp Lys Asn His Asn Trp Asp Thr Arg Glu Val Tyr Pro Thr
340 345 350
Lys Glu Leu Glu Lys Leu Leu Lys Asp Tyr Ser Ile Glu Tyr Gly His
355 360 365
Gly Glu Cys Ile Lys Ala Ala Ile Cys Gly Glu Ser Asp Lys Lys Phe
370 375 380
Phe Ala Lys Leu Thr Ser Ile Leu Asn Ser Ile Leu Gln Met Arg Asn
385 390 395 400
Ser Lys Thr Gly Thr Glu Leu Asp Tyr Leu Ile Ser Pro Val Ala Asp
405 410 415
Val Asn Gly Asn Phe Phe Asp Ser Arg His Ala Pro Lys Asn Met Pro
420 425 430
Gln Asp Ala Asp Ala Asn Gly Ala Tyr His Ile Gly Leu Lys Gly Leu
435 440 445
Met Leu Leu Tyr Arg Ile Lys Asn Asn Gln Asp Gly Lys Lys Leu Asn
450 455 460
Leu Val Ile Lys Asn Glu Glu Tyr Phe Glu Phe Val Gln Asn Arg Asn
465 470 475 480
Lys Ser Ser Lys Ile
485
<210> 25
<211> 939
<212> PRT
<213> 蜃楼弗朗西斯菌亚种
<220>
<221> MISC_FEATURE
<222> (1)..(939)
<223> Genbank AJI56734 Cpf1
<400> 25
Met Asn Leu Tyr Ser Asn Leu Thr Asn Lys Tyr Ser Leu Ser Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Ile Pro Gln Gly Glu Thr Leu Glu Asn Ile Lys
20 25 30
Ala Arg Gly Leu Ile Leu Asp Asp Glu Lys Arg Ala Lys Asp Tyr Lys
35 40 45
Lys Ala Lys Gln Ile Ile Asp Lys Tyr His Gln Phe Phe Ile Glu Glu
50 55 60
Ile Leu Ser Ser Val Cys Ile Ser Glu Asp Leu Leu Gln Asn Tyr Ser
65 70 75 80
Asp Val Tyr Phe Lys Leu Lys Lys Ser Asp Asp Asp Asn Leu Gln Lys
85 90 95
Asp Phe Lys Ser Ala Lys Asp Thr Ile Lys Lys His Ile Ser Arg Tyr
100 105 110
Ile Asn Asp Ser Glu Lys Phe Lys Asn Leu Phe Asn Gln Asn Leu Ile
115 120 125
Asp Ala Lys Lys Gly Gln Glu Ser Asp Leu Ile Leu Trp Leu Lys Gln
130 135 140
Ser Lys Asp Asn Gly Ile Glu Leu Phe Lys Ala Asn Ser Asp Ile Thr
145 150 155 160
Asp Ile Asp Glu Ala Leu Glu Ile Ile Lys Ser Phe Lys Gly Trp Thr
165 170 175
Thr Tyr Phe Lys Gly Phe His Glu Asn Arg Lys Asn Val Tyr Ser Ser
180 185 190
Asp Asp Ile Pro Thr Ser Ile Ile Tyr Arg Ile Val Asp Asp Asn Leu
195 200 205
Pro Lys Phe Ile Glu Asn Lys Ala Lys Tyr Glu Asn Leu Lys Asp Lys
210 215 220
Ala Pro Glu Ala Ile Asn Tyr Glu Gln Ile Lys Lys Asp Leu Ala Glu
225 230 235 240
Glu Leu Thr Phe Asp Ile Asp Tyr Lys Thr Ser Glu Val Asn Gln Arg
245 250 255
Val Phe Ser Leu Asp Glu Val Phe Glu Ile Ala Asn Phe Asn Asn Tyr
260 265 270
Leu Asn Gln Ser Gly Ile Thr Lys Phe Asn Thr Ile Ile Gly Gly Lys
275 280 285
Phe Val Asn Gly Glu Asn Thr Lys Arg Lys Gly Ile Asn Glu Tyr Ile
290 295 300
Asn Leu Tyr Ser Gln Gln Ile Asn Asp Lys Thr Leu Lys Lys Tyr Lys
305 310 315 320
Met Ser Val Leu Phe Lys Gln Ile Leu Ser Asp Thr Glu Ser Lys Ser
325 330 335
Phe Val Ile Asp Lys Leu Glu Asp Asp Ser Asp Val Val Thr Thr Met
340 345 350
Gln Ser Phe Tyr Glu Gln Ile Ala Ala Phe Lys Thr Leu Glu Glu Lys
355 360 365
Ser Ile Lys Glu Thr Leu Ser Leu Leu Phe Asp Asp Leu Lys Ala Gln
370 375 380
Lys Leu Asp Leu Ser Lys Ile Tyr Phe Lys Asn Asp Lys Ser Leu Thr
385 390 395 400
Asp Leu Ser Gln Gln Val Phe Asp Asp Tyr Ser Val Ile Gly Thr Ala
405 410 415
Val Leu Glu Tyr Ile Thr Gln Gln Val Ala Pro Lys Asn Leu Asp Asn
420 425 430
Pro Ser Lys Lys Glu Gln Asp Leu Ile Ala Lys Lys Thr Glu Lys Ala
435 440 445
Lys Tyr Leu Ser Leu Glu Thr Ile Lys Leu Ala Leu Glu Glu Phe Asn
450 455 460
Lys Tyr Arg Asp Ile Asp Lys Gln Cys Arg Phe Glu Glu Ile Phe Ala
465 470 475 480
Ser Phe Ala Asp Ile Pro Val Leu Phe Asp Glu Ile Ala Gln Asn Lys
485 490 495
Asn Asn Leu Ala Gln Ile Ser Ile Lys Tyr Gln Asn Gln Gly Lys Lys
500 505 510
Asp Leu Leu Gln Thr Ser Ala Glu Val Asp Val Lys Ala Ile Lys Asp
515 520 525
Leu Leu Asp Gln Thr Asn Asn Leu Leu His Lys Leu Lys Ile Phe His
530 535 540
Ile Thr Gln Ser Glu Asp Lys Ala Asn Ile Leu Asp Lys Asp Glu His
545 550 555 560
Phe Tyr Leu Val Phe Asp Glu Cys Tyr Phe Glu Leu Ala Asn Ile Val
565 570 575
Ala Leu Tyr Asn Lys Ile Arg Asn Tyr Ile Thr Gln Lys Pro Tyr Ser
580 585 590
Asp Glu Lys Phe Lys Leu Asn Phe Glu Asn Ser Thr Leu Ala Asn Gly
595 600 605
Trp Asp Lys Asn Lys Glu Pro Asp Asn Thr Ala Ile Leu Phe Ile Lys
610 615 620
Asp Asp Lys Tyr Tyr Leu Gly Val Met Asn Lys Lys Asn Asn Lys Ile
625 630 635 640
Phe Asp Asp Lys Ala Ile Lys Glu Asn Lys Gly Glu Gly Tyr Lys Lys
645 650 655
Val Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro Lys Val
660 665 670
Phe Phe Ser Ala Lys Ser Ile Asn Phe Tyr Asn Pro Ser Glu Asp Ile
675 680 685
Leu Arg Ile Arg Asn His Ser Thr His Thr Lys Asn Gly Ser Pro Gln
690 695 700
Lys Gly Tyr Glu Lys Leu Glu Phe Asn Ile Glu Asp Cys Arg Lys Phe
705 710 715 720
Ile Asp Phe Tyr Lys His Ser Ile Ser Arg His Pro Glu Trp Lys Asp
725 730 735
Phe Gly Phe Arg Phe Ser Asp Thr Lys Lys Tyr Asn Ser Ile Asp Glu
740 745 750
Phe Tyr Arg Glu Val Glu Asn Gln Gly Tyr Lys Leu Thr Phe Glu Asn
755 760 765
Ile Ser Glu Ser Tyr Ile Asp Ser Leu Val Asp Glu Gly Lys Leu Tyr
770 775 780
Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Val Tyr Ser Lys Gly Lys
785 790 795 800
Pro Asn Leu His Thr Leu Tyr Trp Lys Ala Leu Phe Asp Glu Arg Asn
805 810 815
Leu Gln Asp Val Val Tyr Lys Leu Asn Gly Glu Ala Glu Leu Phe Tyr
820 825 830
Arg Lys Gln Ser Ile Pro Lys Lys Ile Thr His Pro Ala Lys Glu Ala
835 840 845
Ile Ala Asn Lys Asn Lys Asp Asn Pro Lys Lys Glu Ser Ile Phe Glu
850 855 860
Tyr Asp Leu Ile Lys Asp Lys Arg Phe Thr Glu Asp Lys Phe Phe Phe
865 870 875 880
His Cys Pro Ile Thr Ile Asn Phe Lys Ser Ser Gly Ala Asn Lys Phe
885 890 895
Asn Asp Glu Ile Asn Leu Leu Leu Lys Glu Lys Ala Asn Asp Val His
900 905 910
Ile Leu Ser Ile Asp Arg Gly Glu Arg His Leu Ala Tyr Tyr Thr Leu
915 920 925
Val Asp Gly Lys Gly Asn Ile Ile Cys Lys Asn
930 935
<210> 26
<211> 1261
<212> PRT
<213> 牛眼莫拉氏菌
<220>
<221> MISC_FEATURE
<222> (1)..(1261)
<223> Genbank AKG08867 Cpf1
<400> 26
Met Leu Phe Gln Asp Phe Thr His Leu Tyr Pro Leu Ser Lys Thr Val
1 5 10 15
Arg Phe Glu Leu Lys Pro Ile Gly Lys Thr Leu Glu His Ile His Ala
20 25 30
Lys Asn Phe Leu Asn Gln Asp Glu Thr Met Ala Asp Met Tyr Gln Lys
35 40 45
Val Lys Ala Ile Leu Asp Asp Tyr His Arg Asp Phe Ile Ala Asp Met
50 55 60
Met Gly Glu Val Lys Leu Thr Lys Leu Ala Glu Phe Tyr Asp Val Tyr
65 70 75 80
Leu Lys Phe Arg Lys Asn Pro Lys Asp Asp Gly Leu Gln Lys Gln Leu
85 90 95
Lys Asp Leu Gln Ala Val Leu Arg Lys Glu Ile Val Lys Pro Ile Gly
100 105 110
Asn Gly Gly Lys Tyr Lys Ala Gly Tyr Asp Arg Leu Phe Gly Ala Lys
115 120 125
Leu Phe Lys Asp Gly Lys Glu Leu Gly Asp Leu Ala Lys Phe Val Ile
130 135 140
Ala Gln Glu Gly Glu Ser Ser Pro Lys Leu Ala His Leu Ala His Phe
145 150 155 160
Glu Lys Phe Ser Thr Tyr Phe Thr Gly Phe His Asp Asn Arg Lys Asn
165 170 175
Met Tyr Ser Asp Glu Asp Lys His Thr Ala Ile Ala Tyr Arg Leu Ile
180 185 190
His Glu Asn Leu Pro Arg Phe Ile Asp Asn Leu Gln Ile Leu Ala Thr
195 200 205
Ile Lys Gln Lys His Ser Ala Leu Tyr Asp Gln Ile Ile Asn Glu Leu
210 215 220
Thr Ala Ser Gly Leu Asp Val Ser Leu Ala Ser His Leu Asp Gly Tyr
225 230 235 240
His Lys Leu Leu Thr Gln Glu Gly Ile Thr Ala Tyr Asn Thr Leu Leu
245 250 255
Gly Gly Ile Ser Gly Glu Ala Gly Ser Arg Lys Ile Gln Gly Ile Asn
260 265 270
Glu Leu Ile Asn Ser His His Asn Gln His Cys His Lys Ser Glu Arg
275 280 285
Ile Ala Lys Leu Arg Pro Leu His Lys Gln Ile Leu Ser Asp Gly Met
290 295 300
Gly Val Ser Phe Leu Pro Ser Lys Phe Ala Asp Asp Ser Glu Val Cys
305 310 315 320
Gln Ala Val Asn Glu Phe Tyr Arg His Tyr Ala Asp Val Phe Ala Lys
325 330 335
Val Gln Ser Leu Phe Asp Gly Phe Asp Asp Tyr Gln Lys Asp Gly Ile
340 345 350
Tyr Val Glu Tyr Lys Asn Leu Asn Glu Leu Ser Lys Gln Ala Phe Gly
355 360 365
Asp Phe Ala Leu Leu Gly Arg Val Leu Asp Gly Tyr Tyr Val Asp Val
370 375 380
Val Asn Pro Glu Phe Asn Glu Arg Phe Ala Lys Ala Lys Thr Asp Asn
385 390 395 400
Ala Lys Ala Lys Leu Thr Lys Glu Lys Asp Lys Phe Ile Lys Gly Val
405 410 415
His Ser Leu Ala Ser Leu Glu Gln Ala Ile Glu His Tyr Thr Ala Arg
420 425 430
His Asp Asp Glu Ser Val Gln Ala Gly Lys Leu Gly Gln Tyr Phe Lys
435 440 445
His Gly Leu Ala Gly Val Asp Asn Pro Ile Gln Lys Ile His Asn Asn
450 455 460
His Ser Thr Ile Lys Gly Phe Leu Glu Arg Glu Arg Pro Ala Gly Glu
465 470 475 480
Arg Ala Leu Pro Lys Ile Lys Ser Asp Lys Ser Pro Glu Ile Arg Gln
485 490 495
Leu Lys Glu Leu Leu Asp Asn Ala Leu Asn Val Ala His Phe Ala Lys
500 505 510
Leu Leu Thr Thr Lys Thr Thr Leu His Asn Gln Asp Gly Asn Phe Tyr
515 520 525
Gly Glu Phe Gly Ala Leu Tyr Asp Glu Leu Ala Lys Ile Ala Thr Leu
530 535 540
Tyr Asn Lys Val Arg Asp Tyr Leu Ser Gln Lys Pro Phe Ser Thr Glu
545 550 555 560
Lys Tyr Lys Leu Asn Phe Gly Asn Pro Thr Leu Leu Asn Gly Trp Asp
565 570 575
Leu Asn Lys Glu Lys Asp Asn Phe Gly Val Ile Leu Gln Lys Asp Gly
580 585 590
Cys Tyr Tyr Leu Ala Leu Leu Asp Lys Ala His Lys Lys Val Phe Asp
595 600 605
Asn Ala Pro Asn Thr Gly Lys Ser Val Tyr Gln Lys Met Ile Tyr Lys
610 615 620
Leu Leu Pro Gly Pro Asn Lys Met Leu Pro Lys Val Phe Phe Ala Lys
625 630 635 640
Ser Asn Leu Asp Tyr Tyr Asn Pro Ser Ala Glu Leu Leu Asp Lys Tyr
645 650 655
Ala Gln Gly Thr His Lys Lys Gly Asp Asn Phe Asn Leu Lys Asp Cys
660 665 670
His Ala Leu Ile Asp Phe Phe Lys Ala Gly Ile Asn Lys His Pro Glu
675 680 685
Trp Gln His Phe Gly Phe Lys Phe Ser Pro Thr Ser Ser Tyr Gln Asp
690 695 700
Leu Ser Asp Phe Tyr Arg Glu Val Glu Pro Gln Gly Tyr Gln Val Lys
705 710 715 720
Phe Val Asp Ile Asn Ala Asp Tyr Ile Asn Glu Leu Val Glu Gln Gly
725 730 735
Gln Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Pro Lys Ala
740 745 750
His Gly Lys Pro Asn Leu His Thr Leu Tyr Phe Lys Ala Leu Phe Ser
755 760 765
Glu Asp Asn Leu Val Asn Pro Ile Tyr Lys Leu Asn Gly Glu Ala Glu
770 775 780
Ile Phe Tyr Arg Lys Ala Ser Leu Asp Met Asn Glu Thr Thr Ile His
785 790 795 800
Arg Ala Gly Glu Val Leu Glu Asn Lys Asn Pro Asp Asn Pro Lys Lys
805 810 815
Arg Gln Phe Val Tyr Asp Ile Ile Lys Asp Lys Arg Tyr Thr Gln Asp
820 825 830
Lys Phe Met Leu His Val Pro Ile Thr Met Asn Phe Gly Val Gln Gly
835 840 845
Met Thr Ile Lys Glu Phe Asn Lys Lys Val Asn Gln Ser Ile Gln Gln
850 855 860
Tyr Asp Glu Val Asn Val Ile Gly Ile Asp Arg Gly Glu Arg His Leu
865 870 875 880
Leu Tyr Leu Thr Val Ile Asn Ser Lys Gly Glu Ile Leu Glu Gln Arg
885 890 895
Ser Leu Asn Asp Ile Thr Thr Ala Ser Ala Asn Gly Thr Gln Met Thr
900 905 910
Thr Pro Tyr His Lys Ile Leu Asp Lys Arg Glu Ile Glu Arg Leu Asn
915 920 925
Ala Arg Val Gly Trp Gly Glu Ile Glu Thr Ile Lys Glu Leu Lys Ser
930 935 940
Gly Tyr Leu Ser His Val Val His Gln Ile Ser Gln Leu Met Leu Lys
945 950 955 960
Tyr Asn Ala Ile Val Val Leu Glu Asp Leu Asn Phe Gly Phe Lys Arg
965 970 975
Gly Arg Phe Lys Val Glu Lys Gln Ile Tyr Gln Asn Phe Glu Asn Ala
980 985 990
Leu Ile Lys Lys Leu Asn His Leu Val Leu Lys Asp Lys Ala Asp Asp
995 1000 1005
Glu Ile Gly Ser Tyr Lys Asn Ala Leu Gln Leu Thr Asn Asn Phe
1010 1015 1020
Thr Asp Leu Lys Ser Ile Gly Lys Gln Thr Gly Phe Leu Phe Tyr
1025 1030 1035
Val Pro Ala Trp Asn Thr Ser Lys Ile Asp Pro Glu Thr Gly Phe
1040 1045 1050
Val Asp Leu Leu Lys Pro Arg Tyr Glu Asn Ile Ala Gln Ser Gln
1055 1060 1065
Ala Phe Phe Gly Lys Phe Asp Lys Ile Cys Tyr Asn Ala Asp Arg
1070 1075 1080
Gly Tyr Phe Glu Phe His Ile Asp Tyr Ala Lys Phe Asn Asp Lys
1085 1090 1095
Ala Lys Asn Ser Arg Gln Ile Trp Lys Ile Cys Ser His Gly Asp
1100 1105 1110
Lys Arg Tyr Val Tyr Asp Lys Thr Ala Asn Gln Asn Lys Gly Ala
1115 1120 1125
Thr Ile Gly Val Asn Val Asn Asp Glu Leu Lys Ser Leu Phe Thr
1130 1135 1140
Arg Tyr His Ile Asn Asp Lys Gln Pro Asn Leu Val Met Asp Ile
1145 1150 1155
Cys Gln Asn Asn Asp Lys Glu Phe His Lys Ser Leu Met Tyr Leu
1160 1165 1170
Leu Lys Thr Leu Leu Ala Leu Arg Tyr Ser Asn Ala Ser Ser Asp
1175 1180 1185
Glu Asp Phe Ile Leu Ser Pro Val Ala Asn Asp Glu Gly Val Phe
1190 1195 1200
Phe Asn Ser Ala Leu Ala Asp Asp Thr Gln Pro Gln Asn Ala Asp
1205 1210 1215
Ala Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Leu Trp Leu Leu
1220 1225 1230
Asn Glu Leu Lys Asn Ser Asp Asp Leu Asn Lys Val Lys Leu Ala
1235 1240 1245
Ile Asp Asn Gln Thr Trp Leu Asn Phe Ala Gln Asn Arg
1250 1255 1260
<210> 27
<211> 1282
<212> PRT
<213> 挑剔真杆菌
<220>
<221> MISC_FEATURE
<222> (1)..(1282)
<223> Genbank CDA41776 Cpf1
<400> 27
Met Asn Gly Asn Arg Ser Ile Val Tyr Arg Glu Phe Val Gly Val Thr
1 5 10 15
Pro Val Ala Lys Thr Leu Arg Asn Glu Leu Arg Pro Val Gly His Thr
20 25 30
Gln Glu His Ile Ile Gln Asn Gly Leu Ile Gln Glu Asp Glu Leu Arg
35 40 45
Gln Glu Lys Ser Thr Glu Leu Lys Asn Ile Met Asp Asp Tyr Tyr Arg
50 55 60
Glu Tyr Ile Asp Lys Ser Leu Ser Gly Leu Thr Asp Leu Asp Phe Thr
65 70 75 80
Leu Leu Phe Glu Leu Met Asn Ser Val Gln Ser Ser Leu Ser Lys Asp
85 90 95
Asn Lys Lys Ala Leu Glu Lys Glu His Asn Lys Met Arg Glu Gln Ile
100 105 110
Cys Thr His Leu Gln Ser Asp Ser Asp Tyr Lys Asn Met Phe Asn Ala
115 120 125
Lys Leu Phe Lys Glu Ile Leu Pro Asp Phe Ile Lys Asn Tyr Asn Gln
130 135 140
Tyr Asp Val Lys Asp Lys Ala Gly Lys Leu Glu Thr Leu Ala Leu Phe
145 150 155 160
Asn Gly Phe Ser Thr Tyr Phe Thr Asp Phe Phe Glu Lys Arg Lys Asn
165 170 175
Val Phe Thr Lys Glu Ala Val Ser Thr Ser Ile Ala Tyr Arg Ile Val
180 185 190
His Glu Asn Ser Leu Ile Phe Leu Ala Asn Met Thr Ser Tyr Lys Lys
195 200 205
Ile Ser Glu Lys Ala Leu Asp Glu Ile Glu Val Ile Glu Lys Asn Asn
210 215 220
Gln Asp Lys Met Gly Asp Trp Glu Leu Asn Gln Ile Phe Asn Pro Asp
225 230 235 240
Phe Tyr Asn Met Val Leu Ile Gln Ser Gly Ile Asp Phe Tyr Asn Glu
245 250 255
Ile Cys Gly Val Val Asn Ala His Met Asn Leu Tyr Cys Gln Gln Thr
260 265 270
Lys Asn Asn Tyr Asn Leu Phe Lys Met Arg Lys Leu His Lys Gln Ile
275 280 285
Leu Ala Tyr Thr Ser Thr Ser Phe Glu Val Pro Lys Met Phe Glu Asp
290 295 300
Asp Met Ser Val Tyr Asn Ala Val Asn Ala Phe Ile Asp Glu Thr Glu
305 310 315 320
Lys Gly Asn Ile Ile Gly Lys Leu Lys Asp Ile Val Asn Lys Tyr Asp
325 330 335
Glu Leu Asp Glu Lys Arg Ile Tyr Ile Ser Lys Asp Phe Tyr Glu Thr
340 345 350
Leu Ser Cys Phe Met Ser Gly Asn Trp Asn Leu Ile Thr Gly Cys Val
355 360 365
Glu Asn Phe Tyr Asp Glu Asn Ile His Ala Lys Gly Lys Ser Lys Glu
370 375 380
Glu Lys Val Lys Lys Ala Val Lys Glu Asp Lys Tyr Lys Ser Ile Asn
385 390 395 400
Asp Val Asn Asp Leu Val Glu Lys Tyr Ile Asp Glu Lys Glu Arg Asn
405 410 415
Glu Phe Lys Asn Ser Asn Ala Lys Gln Tyr Ile Arg Glu Ile Ser Asn
420 425 430
Ile Ile Thr Asp Thr Glu Thr Ala His Leu Glu Tyr Asp Glu His Ile
435 440 445
Ser Leu Ile Glu Ser Glu Glu Lys Ala Asp Glu Ile Lys Lys Arg Leu
450 455 460
Asp Met Tyr Met Asn Met Tyr His Trp Val Lys Ala Phe Ile Val Asp
465 470 475 480
Glu Val Leu Asp Arg Asp Glu Met Phe Tyr Ser Asp Ile Asp Asp Ile
485 490 495
Tyr Asn Ile Leu Glu Asn Ile Val Pro Leu Tyr Asn Arg Val Arg Asn
500 505 510
Tyr Val Thr Gln Lys Pro Tyr Thr Ser Lys Lys Ile Lys Leu Asn Phe
515 520 525
Gln Ser Pro Thr Leu Ala Asn Gly Trp Ser Gln Ser Lys Glu Phe Asp
530 535 540
Asn Asn Ala Ile Ile Leu Ile Arg Asp Asn Lys Tyr Tyr Leu Ala Ile
545 550 555 560
Phe Asn Ala Lys Asn Lys Pro Asp Lys Lys Ile Ile Gln Gly Asn Ser
565 570 575
Asp Lys Lys Asn Asp Asn Asp Tyr Lys Lys Met Val Tyr Asn Leu Leu
580 585 590
Pro Gly Ala Asn Lys Met Leu Pro Lys Val Phe Leu Ser Lys Lys Gly
595 600 605
Ile Glu Thr Phe Lys Pro Ser Asp Tyr Ile Ile Ser Gly Tyr Asn Ala
610 615 620
His Lys His Ile Lys Thr Ser Glu Asn Phe Asp Ile Ser Phe Cys Arg
625 630 635 640
Asp Leu Ile Asp Tyr Phe Lys Asn Ser Ile Glu Lys His Ala Glu Trp
645 650 655
Arg Lys Tyr Glu Phe Lys Phe Ser Ala Thr Asp Ser Tyr Asn Asp Ile
660 665 670
Ser Glu Phe Tyr Arg Glu Val Glu Met Gln Gly Tyr Arg Ile Asp Trp
675 680 685
Thr Tyr Ile Ser Glu Ala Asp Ile Asn Lys Leu Asp Glu Glu Gly Lys
690 695 700
Ile Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ala Glu Asn Ser Thr
705 710 715 720
Gly Lys Glu Asn Leu His Thr Met Tyr Phe Lys Asn Ile Phe Ser Glu
725 730 735
Glu Asn Leu Lys Asn Ile Val Ile Lys Leu Asn Gly Gln Ala Glu Leu
740 745 750
Phe Tyr Arg Lys Ala Ser Val Lys Asn Pro Val Lys His Lys Lys Asp
755 760 765
Ser Val Leu Val Asn Lys Thr Tyr Lys Asn Gln Leu Asp Asn Gly Asp
770 775 780
Val Val Arg Ile Pro Ile Pro Asp Asp Ile Tyr Asn Glu Ile Tyr Lys
785 790 795 800
Met Tyr Asn Gly Tyr Ile Lys Glu Ser Asp Leu Ser Glu Ala Ala Lys
805 810 815
Glu Tyr Leu Asp Lys Val Glu Val Arg Thr Ala Gln Lys Asp Ile Val
820 825 830
Lys Asp Tyr Arg Tyr Thr Val Asp Lys Tyr Phe Ile His Thr Pro Ile
835 840 845
Thr Ile Asn Tyr Lys Val Thr Ala Arg Asn Asn Val Asn Asp Met Ala
850 855 860
Val Lys Tyr Ile Ala Gln Asn Asp Asp Ile His Val Ile Gly Ile Asp
865 870 875 880
Arg Gly Glu Arg Asn Leu Ile Tyr Ile Ser Val Ile Asp Ser His Gly
885 890 895
Asn Ile Val Lys Gln Lys Ser Tyr Asn Ile Leu Asn Asn Tyr Asp Tyr
900 905 910
Lys Lys Lys Leu Val Glu Lys Glu Lys Thr Arg Glu Tyr Ala Arg Lys
915 920 925
Asn Trp Lys Ser Ile Gly Asn Ile Lys Glu Leu Lys Glu Gly Tyr Ile
930 935 940
Ser Gly Val Val His Glu Ile Ala Met Leu Met Val Glu Tyr Asn Ala
945 950 955 960
Ile Ile Ala Met Glu Asp Leu Asn Tyr Gly Phe Lys Arg Gly Arg Phe
965 970 975
Lys Val Glu Arg Gln Val Tyr Gln Lys Phe Glu Ser Met Leu Ile Asn
980 985 990
Lys Leu Asn Tyr Phe Ala Ser Lys Gly Lys Ser Val Asp Glu Pro Gly
995 1000 1005
Gly Leu Leu Lys Gly Tyr Gln Leu Thr Tyr Val Pro Asp Asn Ile
1010 1015 1020
Lys Asn Leu Gly Lys Gln Cys Gly Val Ile Phe Tyr Val Pro Ala
1025 1030 1035
Ala Phe Thr Ser Lys Ile Asp Pro Ser Thr Gly Phe Ile Ser Ala
1040 1045 1050
Phe Asn Phe Lys Ser Ile Ser Thr Asn Ala Ser Arg Lys Gln Phe
1055 1060 1065
Phe Met Gln Phe Asp Glu Ile Arg Tyr Cys Ala Glu Lys Asp Met
1070 1075 1080
Phe Ser Phe Gly Phe Asp Tyr Asn Asn Phe Asp Thr Tyr Asn Ile
1085 1090 1095
Thr Met Gly Lys Thr Gln Trp Thr Val Tyr Thr Asn Gly Glu Arg
1100 1105 1110
Leu Gln Ser Glu Phe Asn Asn Ala Arg Arg Thr Gly Lys Thr Lys
1115 1120 1125
Ser Ile Asn Leu Thr Glu Thr Ile Lys Leu Leu Leu Glu Asp Asn
1130 1135 1140
Glu Ile Asn Tyr Ala Asp Gly His Asp Val Arg Ile Asp Met Glu
1145 1150 1155
Lys Met Tyr Glu Asp Lys Asn Ser Glu Phe Phe Ala Gln Leu Leu
1160 1165 1170
Ser Leu Tyr Lys Leu Thr Val Gln Met Arg Asn Ser Tyr Thr Glu
1175 1180 1185
Ala Glu Glu Gln Glu Lys Gly Ile Ser Tyr Asp Lys Ile Ile Ser
1190 1195 1200
Pro Val Ile Asn Asp Glu Gly Glu Phe Phe Asp Ser Asp Asn Tyr
1205 1210 1215
Lys Glu Ser Asp Asp Lys Glu Cys Lys Met Pro Lys Asp Ala Asp
1220 1225 1230
Ala Asn Gly Ala Tyr Cys Ile Ala Leu Lys Gly Leu Tyr Glu Val
1235 1240 1245
Leu Lys Ile Lys Ser Glu Trp Thr Glu Asp Gly Phe Asp Arg Asn
1250 1255 1260
Cys Leu Lys Leu Pro His Ala Glu Trp Leu Asp Phe Ile Gln Asn
1265 1270 1275
Lys Arg Tyr Glu
1280
<210> 28
<211> 1263
<212> PRT
<213> 直肠真杆菌
<220>
<221> MISC_FEATURE
<222> (1)..(1263)
<223> Genbank CUM80100 Cpf1
<400> 28
Met Asn Asn Gly Thr Asn Asn Phe Gln Asn Phe Ile Gly Ile Ser Ser
1 5 10 15
Leu Gln Lys Thr Leu Arg Asn Ala Leu Ile Pro Thr Glu Thr Thr Gln
20 25 30
Gln Phe Ile Val Lys Asn Gly Ile Ile Lys Glu Asp Glu Leu Arg Gly
35 40 45
Glu Asn Arg Gln Ile Leu Lys Asp Ile Met Asp Asp Tyr Tyr Arg Gly
50 55 60
Phe Ile Ser Glu Thr Leu Ser Ser Ile Asp Asp Ile Asp Trp Thr Ser
65 70 75 80
Leu Phe Glu Lys Met Glu Ile Gln Leu Lys Asn Gly Asp Asn Lys Asp
85 90 95
Thr Leu Ile Lys Glu Gln Ala Glu Lys Arg Lys Ala Ile Tyr Lys Lys
100 105 110
Phe Ala Asp Asp Asp Arg Phe Lys Asn Met Phe Ser Ala Lys Leu Ile
115 120 125
Ser Asp Ile Leu Pro Glu Phe Val Ile His Asn Asn Asn Tyr Ser Ala
130 135 140
Ser Glu Lys Glu Glu Lys Thr Gln Val Ile Lys Leu Phe Ser Arg Phe
145 150 155 160
Ala Thr Ser Phe Lys Asp Tyr Phe Lys Asn Arg Ala Asn Cys Phe Ser
165 170 175
Ala Asp Asp Ile Ser Ser Ser Ser Cys His Arg Ile Val Asn Asp Asn
180 185 190
Ala Glu Ile Phe Phe Ser Asn Ala Leu Val Tyr Arg Arg Ile Val Lys
195 200 205
Asn Leu Ser Asn Asp Asp Ile Asn Lys Ile Ser Gly Asp Met Lys Asp
210 215 220
Ser Leu Lys Glu Met Ser Leu Asp Glu Ile Tyr Ser Tyr Glu Lys Tyr
225 230 235 240
Gly Glu Phe Ile Thr Gln Glu Gly Ile Ser Phe Tyr Asn Asp Ile Cys
245 250 255
Gly Lys Val Asn Ser Phe Met Asn Leu Tyr Cys Gln Lys Asn Lys Glu
260 265 270
Asn Lys Asn Leu Tyr Lys Leu Arg Lys Leu His Lys Gln Ile Leu Cys
275 280 285
Ile Ala Asp Thr Ser Tyr Glu Val Pro Tyr Lys Phe Glu Ser Asp Glu
290 295 300
Glu Val Tyr Gln Ser Val Asn Gly Phe Leu Asp Asn Ile Ser Ser Lys
305 310 315 320
His Ile Val Glu Arg Leu Arg Lys Ile Gly Asp Asn Tyr Asn Gly Tyr
325 330 335
Asn Leu Asp Lys Ile Tyr Ile Val Ser Arg Phe Tyr Glu Ser Val Ser
340 345 350
Gln Lys Thr Tyr Arg Asp Trp Glu Thr Ile Asn Thr Ala Leu Glu Ile
355 360 365
His Tyr Asn Asn Ile Leu Pro Gly Asn Gly Lys Ser Lys Ala Asp Lys
370 375 380
Val Lys Lys Ala Val Lys Asn Asp Leu Gln Lys Ser Ile Thr Glu Ile
385 390 395 400
Asn Glu Leu Val Ser Asn Tyr Lys Leu Cys Pro Asp Asp Asn Ile Lys
405 410 415
Ala Glu Thr Tyr Ile His Glu Ile Ser His Ile Leu Asn Asn Phe Glu
420 425 430
Ala Gln Glu Leu Lys Tyr Asn Pro Glu Ile His Leu Val Glu Ser Glu
435 440 445
Leu Lys Ala Ser Glu Leu Lys Asn Val Leu Asp Val Ile Met Asn Ala
450 455 460
Phe His Trp Cys Ser Val Phe Met Thr Glu Glu Leu Val Asp Lys Asp
465 470 475 480
Asn Asn Phe Tyr Ala Glu Leu Glu Glu Ile Tyr Asp Glu Ile Tyr Pro
485 490 495
Val Ile Ser Leu Tyr Asn Leu Val Arg Asn Tyr Val Thr Gln Lys Pro
500 505 510
Tyr Ser Thr Lys Lys Ile Lys Leu Asn Phe Gly Ile Pro Thr Leu Ala
515 520 525
Asp Gly Trp Ser Lys Ser Lys Glu Tyr Ser Asn Asn Ala Ile Ile Leu
530 535 540
Met Arg Asp Asn Leu Tyr Tyr Leu Gly Ile Phe Asn Ala Lys Asn Lys
545 550 555 560
Pro Asp Lys Lys Ile Ile Glu Gly Asn Thr Ser Glu Asn Lys Gly Asp
565 570 575
Tyr Lys Lys Met Ile Tyr Asn Leu Leu Pro Gly Pro Asn Lys Met Ile
580 585 590
Pro Lys Val Phe Leu Ser Ser Lys Thr Gly Val Glu Thr Tyr Lys Pro
595 600 605
Ser Ala Tyr Ile Leu Glu Gly Tyr Lys Gln Asn Lys His Leu Lys Ser
610 615 620
Ser Lys Asp Phe Asp Ile Thr Phe Cys Arg Asp Leu Ile Asp Tyr Phe
625 630 635 640
Lys Asn Cys Ile Ala Ile His Pro Glu Trp Lys Asn Phe Gly Phe Asp
645 650 655
Phe Ser Asp Thr Ser Thr Tyr Glu Asp Ile Ser Gly Phe Tyr Arg Glu
660 665 670
Val Glu Leu Gln Gly Tyr Lys Ile Asp Trp Thr Tyr Ile Ser Glu Lys
675 680 685
Asp Ile Asp Leu Leu Gln Glu Lys Gly Gln Leu Tyr Leu Phe Gln Ile
690 695 700
Tyr Asn Lys Asp Phe Ser Lys Lys Ser Thr Gly Asn Asp Asn Leu His
705 710 715 720
Thr Met Tyr Leu Lys Asn Leu Phe Ser Glu Glu Asn Leu Lys Asp Ile
725 730 735
Val Leu Lys Leu Asn Gly Glu Ala Glu Ile Phe Phe Arg Lys Ser Ser
740 745 750
Ile Lys Asn Pro Ile Ile His Lys Lys Gly Ser Ile Leu Val Asn Arg
755 760 765
Thr Tyr Glu Ala Glu Glu Lys Asp Gln Phe Gly Asn Ile Gln Ile Val
770 775 780
Arg Lys Thr Ile Pro Glu Asn Ile Tyr Gln Glu Leu Tyr Lys Tyr Phe
785 790 795 800
Asn Asp Lys Ser Asp Lys Glu Leu Ser Asp Glu Ala Ala Lys Leu Lys
805 810 815
Asn Val Val Gly His His Glu Ala Ala Thr Asn Ile Val Lys Asp Tyr
820 825 830
Arg Tyr Thr Tyr Asp Lys Tyr Phe Leu His Met Pro Ile Thr Ile Asn
835 840 845
Phe Lys Ala Asn Lys Thr Ser Phe Ile Asn Asp Arg Ile Leu Gln Tyr
850 855 860
Ile Ala Lys Glu Asn Asp Leu His Val Ile Gly Ile Asp Arg Gly Glu
865 870 875 880
Arg Asn Leu Ile Tyr Val Ser Val Ile Asp Thr Cys Gly Asn Ile Val
885 890 895
Glu Gln Lys Ser Phe Asn Ile Val Asn Gly Tyr Asp Tyr Gln Ile Lys
900 905 910
Leu Lys Gln Gln Glu Gly Ala Arg Gln Ile Ala Arg Lys Glu Trp Lys
915 920 925
Glu Ile Gly Lys Ile Lys Glu Ile Lys Glu Gly Tyr Leu Ser Leu Val
930 935 940
Ile His Glu Ile Ser Lys Met Val Ile Lys Tyr Asn Ala Ile Ile Ala
945 950 955 960
Met Glu Asp Leu Ser Tyr Gly Phe Lys Lys Gly Arg Phe Lys Val Glu
965 970 975
Arg Gln Val Tyr Gln Lys Phe Glu Thr Met Leu Ile Asn Lys Leu Asn
980 985 990
Tyr Leu Val Phe Lys Asp Ile Ser Ile Thr Glu Asn Gly Gly Leu Leu
995 1000 1005
Lys Gly Tyr Gln Leu Thr Tyr Ile Pro Glu Lys Leu Lys Asn Val
1010 1015 1020
Gly His Gln Cys Gly Cys Ile Phe Tyr Val Pro Ala Ala Tyr Thr
1025 1030 1035
Ser Lys Ile Asp Pro Thr Thr Gly Phe Ala Asn Ile Phe Lys Phe
1040 1045 1050
Lys Asp Leu Thr Val Asp Ala Lys Arg Glu Phe Ile Lys Lys Phe
1055 1060 1065
Asp Ser Ile Arg Tyr Asp Ser Glu Lys Asn Leu Phe Cys Phe Thr
1070 1075 1080
Phe Asp Tyr Asn Asn Phe Ile Thr Gln Asn Thr Val Met Ser Lys
1085 1090 1095
Ser Ser Trp Ser Val Tyr Thr Tyr Gly Val Arg Ile Lys Arg Arg
1100 1105 1110
Phe Val Asn Gly Arg Phe Ser Asn Glu Ser Asp Thr Ile Asp Ile
1115 1120 1125
Thr Lys Asp Met Glu Lys Thr Leu Glu Met Thr Asp Ile Asn Trp
1130 1135 1140
Arg Asp Gly His Asp Leu Arg Gln Asp Ile Ile Asp Tyr Glu Ile
1145 1150 1155
Val Gln His Ile Phe Glu Ile Phe Lys Leu Thr Val Gln Met Arg
1160 1165 1170
Asn Ser Leu Ser Glu Leu Glu Asp Arg Asp Tyr Asp Arg Leu Ile
1175 1180 1185
Ser Pro Val Leu Asn Glu Asn Asn Ile Phe Tyr Asp Ser Ala Lys
1190 1195 1200
Ala Gly Asp Ala Leu Pro Lys Asp Ala Asp Ala Asn Gly Ala Tyr
1205 1210 1215
Cys Ile Ala Leu Lys Gly Leu Tyr Glu Ile Lys Gln Ile Thr Glu
1220 1225 1230
Asn Trp Lys Glu Asp Gly Lys Phe Ser Arg Asp Lys Leu Lys Ile
1235 1240 1245
Ser Asn Lys Asp Trp Phe Asp Phe Ile Gln Asn Lys Arg Tyr Leu
1250 1255 1260
<210> 29
<211> 966
<212> PRT
<213> 未知
<220>
<223> 未培养细菌(地下水宏基因组)假定蛋白ACD_18C00234G0001
<220>
<221> MISC_FEATURE
<222> (1)..(966)
<223> Genbank EKE06926 Cpf1
<400> 29
Met Glu Asn Tyr Asp Lys Cys Leu Thr Gln Leu Gly Ile Glu Lys Phe
1 5 10 15
Asn Leu Glu Ile Val Gly Glu Ile Asn Lys Glu Leu Asn Leu Phe Ser
20 25 30
Gln Gln Asn Arg Asp Ile Leu Pro Lys Ala Pro Lys Leu Lys Phe Leu
35 40 45
Tyr Lys Gln Ile Gly Cys Gly Lys Arg Ile Phe Asp Leu Phe Ser Ile
50 55 60
Ile Val Gly Asn Glu Trp Lys Glu Leu Lys Asn Leu Gln Asn Asn Lys
65 70 75 80
Asp Asp Gly Lys Ser Phe Ser Gln Lys Glu Leu Leu Glu Lys Ile Arg
85 90 95
Lys Leu Tyr Lys Leu Phe Phe Asp Lys Pro Ser Asp Tyr Glu Leu Asp
100 105 110
Lys Ile Tyr Phe Asn Lys Gln Ser Ile Asn Thr Ile Ser Ser Met Trp
115 120 125
Phe Val Asn Trp His Lys Leu Ser Glu Leu Leu Ser Gly Lys Arg Ile
130 135 140
Ile Lys Asn Lys Asn Lys Glu Thr Gly Glu Tyr Thr Ile Pro Lys Lys
145 150 155 160
Ile Ser Leu Ala Asp Leu Lys Asn Ile Leu Glu Ser Glu Thr Asn Val
165 170 175
Glu Asp Leu Phe Lys Lys Gly Lys Ile Asn Glu Asp Glu Ile Lys Glu
180 185 190
Asn Asn Ser Val Gly Val Tyr Glu Lys Leu Phe Ser Ser Asn Gly Trp
195 200 205
Glu Thr Phe Leu Ala Ile Trp Glu Tyr Glu Ile Asn Glu Ser Phe Lys
210 215 220
Val Leu Asp Asn Asn Val Ala Lys Phe Glu Leu Lys Lys Gln Thr Lys
225 230 235 240
Phe Gln Asn Leu Asp Lys Lys Glu Arg Val Phe Phe Ile Lys Glu Phe
245 250 255
Cys Asp Ala Phe Leu Ala Ile Glu Arg Met Val Lys Tyr His Lys Val
260 265 270
Asp Glu Asn Asn Asp Thr Asp Asp Asn Phe Tyr Glu Thr Ile Asp Leu
275 280 285
Tyr Leu Gln Glu Thr Glu Leu Arg Lys Tyr Tyr Asp Ala Phe Arg Asn
290 295 300
Tyr Leu Thr Glu Lys Pro Phe Ser Glu Asn Lys Ile Lys Leu Asn Phe
305 310 315 320
Lys Ser Gly Thr Leu Leu Gly Gly Trp Ser Gln Thr Phe Glu Thr Tyr
325 330 335
Gly Ser Leu Ile Phe Glu Lys Asn Gly Glu Tyr Phe Leu Gly Ile Ile
340 345 350
Asn Gly Thr Lys Phe Ser Glu Ser Glu Leu Asn Lys Ile Tyr Asn Ile
355 360 365
Asn Ser Asp Ser Ile Lys Ala Lys Arg Leu Leu Tyr Asn Thr Gln Lys
370 375 380
Ile Asp Asn Lys Asn Pro Pro Arg Trp Phe Ile Arg Ser Lys Lys Thr
385 390 395 400
Thr Phe Ser Pro Met Val Arg Glu Gly Leu Leu Asp Pro Glu Ser Ile
405 410 415
Leu Glu Leu Tyr Asp Lys Lys Leu Tyr Ser Lys Thr Glu Asn Lys Asn
420 425 430
Gly Tyr Lys Glu Tyr Leu Pro Arg Leu Leu Asp Tyr Phe Lys Asp Gly
435 440 445
Phe Leu Lys His Lys Asp Phe Val Gln Phe Lys Glu Asn Phe Lys Trp
450 455 460
Leu Asp Asn Ser Glu Tyr Asp Thr Val Val Asp Phe Tyr Asn His Thr
465 470 475 480
Ala Asp Met Cys Tyr Lys Thr Ser Trp Glu Asp Ile Asn Phe Thr Glu
485 490 495
Leu Glu Asn Leu Thr Lys Asp Ser Arg Ile Tyr Leu Phe Lys Ile Tyr
500 505 510
Asn Lys Asp Phe Ala Glu Lys Thr Ser Gly Ile Lys Asn Ser His Thr
515 520 525
Ile Leu Phe Leu Glu Leu Leu Lys Ser Glu Asn Asn Leu Lys Leu Lys
530 535 540
Leu Leu Gly Gly Gly Glu Val Phe Tyr Arg Glu Lys Ser Ile Glu Lys
545 550 555 560
Glu Ile Asp Lys Glu Arg Ser Phe Lys Thr Asp Lys Phe Glu Ile Ile
565 570 575
Lys Asn Lys Arg Tyr Ser Glu Glu Lys Tyr Phe Leu His Phe Pro Ile
580 585 590
Glu Ile Lys Gly Arg Lys Leu Lys Gly Ser Phe Asn Gln Phe Leu Asn
595 600 605
Lys Glu Ile Ser Lys Lys Glu Ser Val Asn Ile Leu Gly Ile Asp Arg
610 615 620
Gly Glu Lys His Leu Leu Tyr Tyr Ser Leu Val Asn Asn Asn Gly Glu
625 630 635 640
Ile Ile Lys Gln Gly Ser Phe Asn Lys Ile Lys Cys Gly Asn Lys Ile
645 650 655
Val Asp Tyr Asn Glu Leu Leu Ser Lys Arg Ala Lys Glu Met Met Glu
660 665 670
Ala Arg Gln Ser Trp Glu Thr Ile Gly Lys Ile Lys Asp Leu Lys Glu
675 680 685
Gly Tyr Leu Ser Gln Val Ile His Glu Ile Tyr Lys Leu Val Ile Glu
690 695 700
Asn Asn Ala Ile Val Val Leu Glu Asp Leu Asn Thr Glu Phe Lys Ala
705 710 715 720
Lys Arg Thr Ala Lys Val Glu Lys Ser Val Tyr Lys Lys Phe Glu Leu
725 730 735
Ala Leu Val Lys Lys Leu Asn His Leu Ile Leu Lys Glu Lys Lys Ala
740 745 750
Asn Glu Leu Gly Gly Ser Leu Asn Ala Tyr Gln Leu Thr Pro Tyr Ile
755 760 765
Lys Pro Gly Asp Val Asp Lys Phe Glu Lys Ala Lys Gln Trp Gly Ile
770 775 780
Met Phe Tyr Val Arg Pro Asp Tyr Thr Ser Gln Thr Asp Pro Val Thr
785 790 795 800
Gly Trp Arg Lys Thr Ile Tyr Ile Ser Asn Ser Glu Thr Ile Glu Asn
805 810 815
Ile Lys Lys Lys Trp Lys Asp Ala Asn Ile Lys Ile Tyr Phe Asp Ser
820 825 830
Asp Lys Lys Cys Phe Lys Phe Leu Tyr Asp Lys Trp Glu Leu Cys Ala
835 840 845
Tyr Pro Asn Leu Glu Arg Leu Tyr Trp Asn Arg Ser Glu Lys Asn Ala
850 855 860
Glu Gly Lys Phe Gly Asn Met Lys Lys Tyr Ser Leu His Lys Glu Phe
865 870 875 880
Glu Cys Ile Phe Glu Gly Val Asn Lys Ser Lys Asn Ile Ser Asp Gln
885 890 895
Met Phe Asp Lys Glu Asp Phe Asn Trp Lys Ser Phe Ile Phe Tyr Trp
900 905 910
Asn Leu Leu Asn Gln Ile Arg Asn Ser Asp Lys Ser Lys Asp Glu Asn
915 920 925
Glu Ser Asp Phe Ile Gln Ser Pro Ile Trp Ser Glu Lys Ile Asn Asp
930 935 940
Phe Phe Asp Ser Arg Lys Lys Tyr Gln Ile Asp Leu Pro Glu Asn Gly
945 950 955 960
Asp Ala Asn Gly Ala Tyr
965
<210> 30
<211> 1247
<212> PRT
<213> 未知
<220>
<223> 未培养细菌(gcode 4)(地下水宏基因组)假定蛋白ACD_3C00058G0015
<220>
<221> MISC_FEATURE
<222> (1)..(1247)
<223> Genbank EKE28449 Cpf1
<400> 30
Met Phe Lys Gly Asp Ala Phe Thr Gly Leu Tyr Glu Val Gln Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Val Pro Ile Gly Leu Thr Gln Ser Tyr Leu Glu
20 25 30
Asn Asp Trp Val Ile Gln Lys Asp Lys Glu Val Glu Glu Asn Tyr Gly
35 40 45
Lys Ile Lys Ala Tyr Phe Asp Leu Ile His Lys Glu Phe Val Arg Gln
50 55 60
Ser Leu Glu Asn Ala Trp Leu Cys Gln Leu Asp Asp Phe Tyr Glu Lys
65 70 75 80
Tyr Ile Glu Leu His Asn Ser Leu Glu Thr Arg Lys Asp Lys Asn Leu
85 90 95
Ala Lys Gln Phe Glu Lys Val Met Lys Ser Leu Lys Lys Glu Phe Val
100 105 110
Ser Phe Phe Asp Ala Lys Trp Asn Glu Trp Lys Gln Lys Phe Ser Phe
115 120 125
Leu Lys Lys Trp Trp Ile Asp Val Leu Asn Glu Lys Glu Val Leu Asp
130 135 140
Leu Met Ala Glu Phe Tyr Pro Asp Glu Lys Glu Leu Phe Asp Lys Phe
145 150 155 160
Asp Lys Phe Phe Thr Tyr Phe Ser Asn Phe Lys Glu Ser Arg Lys Asn
165 170 175
Phe Tyr Ala Asp Asp Gly Arg Ala Trp Ala Ile Ala Thr Arg Ala Ile
180 185 190
Asp Glu Asn Leu Ile Thr Phe Ile Lys Asn Ile Glu Asp Phe Lys Lys
195 200 205
Leu Asn Ser Ser Phe Arg Glu Phe Val Asn Asp Asn Phe Ser Glu Glu
210 215 220
Asp Lys Gln Ile Phe Glu Ile Asp Phe Tyr Asn Asn Cys Leu Leu Gln
225 230 235 240
Pro Trp Ile Asp Lys Tyr Asn Lys Ile Val Trp Trp Tyr Ser Leu Glu
245 250 255
Asn Trp Glu Lys Val Gln Trp Leu Asn Glu Lys Ile Asn Asn Phe Lys
260 265 270
Gln Asn Gln Asn Lys Ser Asn Ser Lys Asp Leu Lys Phe Pro Arg Met
275 280 285
Lys Leu Leu Tyr Lys Gln Ile Leu Gly Asp Lys Glu Lys Lys Val Tyr
290 295 300
Ile Asp Glu Ile Arg Asp Asp Lys Asn Leu Ile Asp Leu Ile Asp Asn
305 310 315 320
Ser Lys Arg Arg Asn Gln Ile Lys Ile Asp Asn Ala Asn Asp Ile Ile
325 330 335
Asn Asp Phe Ile Asn Asn Asn Ala Lys Phe Glu Leu Asp Lys Ile Tyr
340 345 350
Leu Thr Arg Gln Ser Ile Asn Thr Ile Ser Ser Lys Tyr Phe Ser Ser
355 360 365
Trp Asp Tyr Ile Arg Trp Tyr Phe Trp Thr Gly Glu Leu Gln Glu Phe
370 375 380
Val Ser Phe Tyr Asp Leu Lys Glu Thr Phe Trp Lys Ile Glu Tyr Glu
385 390 395 400
Thr Leu Glu Asn Ile Phe Lys Asp Cys Tyr Val Lys Gly Ile Asn Thr
405 410 415
Glu Ser Gln Asn Asn Ile Val Phe Glu Thr Gln Gly Ile Tyr Glu Asn
420 425 430
Phe Leu Asn Ile Phe Lys Phe Glu Phe Asn Gln Asn Ile Ser Gln Ile
435 440 445
Ser Leu Leu Glu Trp Glu Leu Asp Lys Ile Gln Asn Glu Asp Ile Lys
450 455 460
Lys Asn Glu Lys Gln Val Glu Val Ile Lys Asn Tyr Phe Asp Ser Val
465 470 475 480
Met Ser Val Tyr Lys Met Thr Lys Tyr Phe Ser Leu Glu Lys Trp Lys
485 490 495
Lys Arg Val Glu Leu Asp Thr Asp Asn Asn Phe Tyr Asn Asp Phe Asn
500 505 510
Glu Tyr Leu Glu Gly Phe Glu Ile Trp Lys Asp Tyr Asn Leu Val Arg
515 520 525
Asn Tyr Ile Thr Lys Lys Gln Val Asn Thr Asp Lys Ile Lys Leu Asn
530 535 540
Phe Asp Asn Ser Gln Phe Leu Thr Trp Trp Asp Lys Asp Lys Glu Asn
545 550 555 560
Glu Arg Leu Gly Ile Ile Leu Arg Arg Glu Trp Lys Tyr Tyr Leu Trp
565 570 575
Ile Leu Lys Lys Trp Asn Thr Leu Asn Phe Gly Asp Tyr Leu Gln Lys
580 585 590
Glu Trp Glu Ile Phe Tyr Glu Lys Met Asn Tyr Lys Gln Leu Asn Asn
595 600 605
Val Tyr Arg Gln Leu Pro Arg Leu Leu Phe Pro Leu Thr Lys Lys Leu
610 615 620
Asn Glu Leu Lys Trp Asp Glu Leu Lys Lys Tyr Leu Ser Lys Tyr Ile
625 630 635 640
Gln Asn Phe Trp Tyr Asn Glu Glu Ile Ala Gln Ile Lys Ile Glu Phe
645 650 655
Asp Ile Phe Gln Glu Ser Lys Glu Lys Trp Glu Lys Phe Asp Ile Asp
660 665 670
Lys Leu Arg Lys Leu Ile Glu Tyr Tyr Lys Lys Trp Val Leu Ala Leu
675 680 685
Tyr Ser Asp Leu Tyr Asp Leu Glu Phe Ile Lys Tyr Lys Asn Tyr Asp
690 695 700
Asp Leu Ser Ile Phe Tyr Ser Asp Val Glu Lys Lys Met Tyr Asn Leu
705 710 715 720
Asn Phe Thr Lys Ile Asp Lys Ser Leu Ile Asp Gly Lys Val Lys Ser
725 730 735
Trp Glu Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Glu Ser
740 745 750
Lys Lys Glu Trp Ser Thr Glu Asn Ile His Thr Lys Tyr Phe Lys Leu
755 760 765
Leu Phe Asn Glu Lys Asn Leu Gln Asn Leu Val Val Lys Leu Ser Trp
770 775 780
Trp Ala Asp Ile Phe Phe Arg Asp Lys Thr Glu Asn Leu Lys Phe Lys
785 790 795 800
Lys Asp Lys Asn Gly Gln Glu Ile Leu Asp His Arg Arg Phe Ser Gln
805 810 815
Asp Lys Ile Met Phe His Ile Ser Ile Thr Leu Asn Ala Asn Cys Trp
820 825 830
Asp Lys Tyr Trp Phe Asn Gln Tyr Val Asn Glu Tyr Met Asn Lys Glu
835 840 845
Arg Asp Ile Lys Ile Ile Trp Ile Asp Arg Trp Glu Lys His Leu Ala
850 855 860
Tyr Tyr Cys Val Ile Asp Lys Ser Trp Lys Ile Phe Asn Asn Glu Ile
865 870 875 880
Trp Thr Leu Asn Glu Leu Asn Trp Val Asn Tyr Leu Glu Lys Leu Glu
885 890 895
Lys Ile Glu Ser Ser Arg Lys Asp Ser Arg Ile Ser Trp Trp Glu Ile
900 905 910
Glu Asn Ile Lys Glu Leu Lys Asn Gly Tyr Ile Ser Gln Val Ile Asn
915 920 925
Lys Leu Thr Glu Leu Ile Val Lys Tyr Asn Ala Ile Ile Val Phe Glu
930 935 940
Asp Leu Asn Ile Trp Phe Lys Arg Trp Arg Gln Lys Ile Glu Lys Gln
945 950 955 960
Ile Tyr Gln Lys Leu Glu Leu Ala Leu Ala Lys Lys Leu Asn Tyr Leu
965 970 975
Thr Gln Lys Asp Lys Lys Asp Asp Glu Ile Leu Trp Asn Leu Lys Ala
980 985 990
Leu Gln Leu Val Pro Lys Val Asn Asp Tyr Gln Asp Ile Trp Asn Tyr
995 1000 1005
Lys Gln Ser Trp Ile Met Phe Tyr Val Arg Ala Asn Tyr Thr Ser
1010 1015 1020
Val Thr Cys Pro Asn Cys Trp Leu Arg Lys Asn Leu Tyr Ile Ser
1025 1030 1035
Asn Ser Ala Thr Lys Glu Asn Gln Lys Lys Ser Leu Asn Ser Ile
1040 1045 1050
Ala Ile Lys Tyr Asn Asp Trp Lys Phe Ser Phe Ser Tyr Glu Ile
1055 1060 1065
Asp Asp Lys Ser Trp Lys Gln Lys Gln Ser Leu Asn Lys Lys Lys
1070 1075 1080
Phe Ile Val Tyr Ser Asp Ile Glu Arg Phe Val Tyr Ser Pro Leu
1085 1090 1095
Glu Lys Leu Thr Lys Val Ile Asp Val Asn Lys Lys Leu Leu Glu
1100 1105 1110
Leu Phe Arg Asp Phe Asn Leu Ser Leu Asp Ile Asn Lys Gln Ile
1115 1120 1125
Gln Glu Lys Asp Leu Asp Ser Val Phe Phe Lys Ser Leu Thr His
1130 1135 1140
Leu Phe Asn Leu Ile Leu Gln Leu Arg Asn Ser Asp Ser Lys Asp
1145 1150 1155
Asn Lys Asp Tyr Ile Ser Cys Pro Ser Cys Tyr Tyr His Ser Asn
1160 1165 1170
Asn Trp Leu Gln Trp Phe Glu Phe Asn Trp Asp Ala Asn Trp Ala
1175 1180 1185
Tyr Asn Ile Ala Arg Lys Gly Ile Ile Leu Leu Asp Arg Ile Arg
1190 1195 1200
Lys Asn Gln Glu Lys Pro Asp Leu Tyr Val Ser Asp Ile Asp Trp
1205 1210 1215
Asp Asn Phe Val Gln Ser Asn Gln Phe Pro Asn Thr Ile Ile Pro
1220 1225 1230
Ile Gln Asn Ile Glu Lys Gln Val Pro Leu Asn Ile Lys Ile
1235 1240 1245
<210> 31
<211> 1250
<212> PRT
<213> 史密斯氏菌
<220>
<221> MISC_FEATURE
<222> (1)..(1250)
<223> Genbank KFO67989 Cpf1
<400> 31
Met Gln Thr Leu Phe Glu Asn Phe Thr Asn Gln Tyr Pro Val Ser Lys
1 5 10 15
Thr Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Lys Asp Phe Ile
20 25 30
Glu Gln Lys Gly Leu Leu Lys Lys Asp Glu Asp Arg Ala Glu Lys Tyr
35 40 45
Lys Lys Val Lys Asn Ile Ile Asp Glu Tyr His Lys Asp Phe Ile Glu
50 55 60
Lys Ser Leu Asn Gly Leu Lys Leu Asp Gly Leu Glu Glu Tyr Lys Thr
65 70 75 80
Leu Tyr Leu Lys Gln Glu Lys Asp Asp Lys Asp Lys Lys Ala Phe Asp
85 90 95
Lys Glu Lys Glu Asn Leu Arg Lys Gln Ile Ala Asn Ala Phe Arg Asn
100 105 110
Asn Glu Lys Phe Lys Thr Leu Phe Ala Lys Glu Leu Ile Lys Asn Asp
115 120 125
Leu Met Ser Phe Ala Cys Glu Glu Asp Lys Lys Asn Val Lys Glu Phe
130 135 140
Glu Ala Phe Thr Thr Tyr Phe Thr Gly Phe His Gln Asn Arg Ala Asn
145 150 155 160
Met Tyr Val Ala Asp Glu Lys Arg Thr Ala Ile Ala Ser Arg Leu Ile
165 170 175
His Glu Asn Leu Pro Lys Phe Ile Asp Asn Ile Lys Ile Phe Glu Lys
180 185 190
Met Lys Lys Glu Ala Pro Glu Leu Leu Ser Pro Phe Asn Gln Thr Leu
195 200 205
Lys Asp Met Lys Asp Val Ile Lys Gly Thr Thr Leu Glu Glu Ile Phe
210 215 220
Ser Leu Asp Tyr Phe Asn Lys Thr Leu Thr Gln Ser Gly Ile Asp Ile
225 230 235 240
Tyr Asn Ser Val Ile Gly Gly Arg Thr Pro Glu Glu Gly Lys Thr Lys
245 250 255
Ile Lys Gly Leu Asn Glu Tyr Ile Asn Thr Asp Phe Asn Gln Lys Gln
260 265 270
Thr Asp Lys Lys Lys Arg Gln Pro Lys Phe Lys Gln Leu Tyr Lys Gln
275 280 285
Ile Leu Ser Asp Arg Gln Ser Leu Ser Phe Ile Ala Glu Ala Phe Lys
290 295 300
Asn Asp Thr Glu Ile Leu Glu Ala Ile Glu Lys Phe Tyr Val Asn Glu
305 310 315 320
Leu Leu His Phe Ser Asn Glu Gly Lys Ser Thr Asn Val Leu Asp Ala
325 330 335
Ile Lys Asn Ala Val Ser Asn Leu Glu Ser Phe Asn Leu Thr Lys Ile
340 345 350
Tyr Phe Arg Ser Gly Thr Ser Leu Thr Asp Val Ser Arg Lys Val Phe
355 360 365
Gly Glu Trp Ser Ile Ile Asn Arg Ala Leu Asp Asn Tyr Tyr Ala Thr
370 375 380
Thr Tyr Pro Ile Lys Pro Arg Glu Lys Ser Glu Lys Tyr Glu Glu Arg
385 390 395 400
Lys Glu Lys Trp Leu Lys Gln Asp Phe Asn Val Ser Leu Ile Gln Thr
405 410 415
Ala Ile Asp Glu Tyr Asp Asn Glu Thr Val Lys Gly Lys Asn Ser Gly
420 425 430
Lys Val Ile Val Asp Tyr Phe Ala Lys Phe Cys Asp Asp Lys Glu Thr
435 440 445
Asp Leu Ile Gln Lys Val Asn Glu Gly Tyr Ile Ala Val Lys Asp Leu
450 455 460
Leu Asn Thr Pro Tyr Pro Glu Asn Glu Lys Leu Gly Ser Asn Lys Asp
465 470 475 480
Gln Val Lys Gln Ile Lys Ala Phe Met Asp Ser Ile Met Asp Ile Met
485 490 495
His Phe Val Arg Pro Leu Ser Leu Lys Asp Thr Asp Lys Glu Lys Asp
500 505 510
Glu Thr Phe Tyr Ser Leu Phe Thr Pro Leu Tyr Asp His Leu Thr Gln
515 520 525
Thr Ile Ala Leu Tyr Asn Lys Val Arg Asn Tyr Leu Thr Gln Lys Pro
530 535 540
Tyr Ser Thr Glu Lys Ile Lys Leu Asn Phe Glu Asn Ser Thr Leu Leu
545 550 555 560
Gly Gly Trp Asp Leu Asn Lys Glu Thr Asp Asn Thr Ala Ile Ile Leu
565 570 575
Arg Lys Glu Asn Leu Tyr Tyr Leu Gly Ile Met Asp Lys Arg His Asn
580 585 590
Arg Ile Phe Arg Asn Val Pro Lys Ala Asp Lys Lys Asp Ser Cys Tyr
595 600 605
Glu Lys Met Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro
610 615 620
Lys Val Phe Phe Ser Gln Ser Arg Ile Gln Glu Phe Thr Pro Ser Ala
625 630 635 640
Lys Leu Leu Glu Asn Tyr Glu Asn Glu Thr His Lys Lys Gly Asp Asn
645 650 655
Phe Asn Leu Asn His Cys His Gln Leu Ile Asp Phe Phe Lys Asp Ser
660 665 670
Ile Asn Lys His Glu Asp Trp Lys Asn Phe Asp Phe Arg Phe Ser Ala
675 680 685
Thr Ser Thr Tyr Ala Asp Leu Ser Gly Phe Tyr His Glu Val Glu His
690 695 700
Gln Gly Tyr Lys Ile Ser Phe Gln Ser Ile Ala Asp Ser Phe Ile Asp
705 710 715 720
Asp Leu Val Asn Glu Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys
725 730 735
Asp Phe Ser Pro Phe Ser Lys Gly Lys Pro Asn Leu His Thr Leu Tyr
740 745 750
Trp Lys Met Leu Phe Asp Glu Asn Asn Leu Lys Asp Val Val Tyr Lys
755 760 765
Leu Asn Gly Glu Ala Glu Val Phe Tyr Arg Lys Lys Ser Ile Ala Glu
770 775 780
Lys Asn Thr Thr Ile His Lys Ala Asn Glu Ser Ile Ile Asn Lys Asn
785 790 795 800
Pro Asp Asn Pro Lys Ala Thr Ser Thr Phe Asn Tyr Asp Ile Val Lys
805 810 815
Asp Lys Arg Tyr Thr Ile Asp Lys Phe Gln Phe His Val Pro Ile Thr
820 825 830
Met Asn Phe Lys Ala Glu Gly Ile Phe Asn Met Asn Gln Arg Val Asn
835 840 845
Gln Phe Leu Lys Ala Asn Pro Asp Ile Asn Ile Ile Gly Ile Asp Arg
850 855 860
Gly Glu Arg His Leu Leu Tyr Tyr Thr Leu Ile Asn Gln Lys Gly Lys
865 870 875 880
Ile Leu Lys Gln Asp Thr Leu Asn Val Ile Ala Asn Glu Lys Gln Lys
885 890 895
Val Asp Tyr His Asn Leu Leu Asp Lys Lys Glu Gly Asp Arg Ala Thr
900 905 910
Ala Arg Gln Glu Trp Gly Val Ile Glu Thr Ile Lys Glu Leu Lys Glu
915 920 925
Gly Tyr Leu Ser Gln Val Ile His Lys Leu Thr Asp Leu Met Ile Glu
930 935 940
Asn Asn Ala Ile Ile Val Met Glu Asp Leu Asn Phe Gly Phe Lys Arg
945 950 955 960
Gly Arg Gln Lys Val Glu Lys Gln Val Tyr Gln Lys Phe Glu Lys Met
965 970 975
Leu Ile Asp Lys Leu Asn Tyr Leu Val Asp Lys Asn Lys Lys Ala Asn
980 985 990
Glu Leu Gly Gly Leu Leu Asn Ala Phe Gln Leu Ala Asn Lys Phe Glu
995 1000 1005
Ser Phe Gln Lys Met Gly Lys Gln Asn Gly Phe Ile Phe Tyr Val
1010 1015 1020
Pro Ala Trp Asn Thr Ser Lys Thr Asp Pro Ala Thr Gly Phe Ile
1025 1030 1035
Asp Phe Leu Lys Pro Arg Tyr Glu Asn Leu Lys Gln Ala Lys Asp
1040 1045 1050
Phe Phe Glu Lys Phe Asp Ser Ile Arg Leu Asn Ser Lys Ala Asp
1055 1060 1065
Tyr Phe Glu Phe Ala Phe Asp Phe Lys Asn Phe Thr Gly Lys Ala
1070 1075 1080
Asp Gly Gly Arg Thr Lys Trp Thr Val Cys Thr Thr Asn Glu Asp
1085 1090 1095
Arg Tyr Ala Trp Asn Arg Ala Leu Asn Asn Asn Arg Gly Ser Gln
1100 1105 1110
Glu Lys Tyr Asp Ile Thr Ala Glu Leu Lys Ser Leu Phe Asp Gly
1115 1120 1125
Lys Val Asp Tyr Lys Ser Gly Lys Asp Leu Lys Gln Gln Ile Ala
1130 1135 1140
Ser Gln Glu Leu Ala Asp Phe Phe Arg Thr Leu Met Lys Tyr Leu
1145 1150 1155
Ser Val Thr Leu Ser Leu Arg His Asn Asn Gly Glu Lys Gly Glu
1160 1165 1170
Thr Glu Gln Asp Tyr Ile Leu Ser Pro Val Ala Asp Ser Met Gly
1175 1180 1185
Lys Phe Phe Asp Ser Arg Lys Ala Gly Asp Asp Met Pro Lys Asn
1190 1195 1200
Ala Asp Ala Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Leu Trp
1205 1210 1215
Cys Leu Glu Gln Ile Ser Lys Thr Asp Asp Leu Lys Lys Val Lys
1220 1225 1230
Leu Ala Ile Ser Asn Lys Glu Trp Leu Glu Phe Met Gln Thr Leu
1235 1240 1245
Lys Gly
1250
<210> 32
<211> 1477
<212> PRT
<213> 异域菌门暂定种
<220>
<221> MISC_FEATURE
<222> (1)..(1477)
<223> Genbank KKP36646 Cpf1
<400> 32
Met Ser Asn Phe Phe Lys Asn Phe Thr Asn Leu Tyr Glu Leu Ser Lys
1 5 10 15
Thr Leu Arg Phe Glu Leu Lys Pro Val Gly Asp Thr Leu Thr Asn Met
20 25 30
Lys Asp His Leu Glu Tyr Asp Glu Lys Leu Gln Thr Phe Leu Lys Asp
35 40 45
Gln Asn Ile Asp Asp Ala Tyr Gln Ala Leu Lys Pro Gln Phe Asp Glu
50 55 60
Ile His Glu Glu Phe Ile Thr Asp Ser Leu Glu Ser Lys Lys Ala Lys
65 70 75 80
Glu Ile Asp Phe Ser Glu Tyr Leu Asp Leu Phe Gln Glu Lys Lys Glu
85 90 95
Leu Asn Asp Ser Glu Lys Lys Leu Arg Asn Lys Ile Gly Glu Thr Phe
100 105 110
Asn Lys Ala Gly Glu Lys Trp Lys Lys Glu Lys Tyr Pro Gln Tyr Glu
115 120 125
Trp Lys Lys Gly Ser Lys Ile Ala Asn Gly Ala Asp Ile Leu Ser Cys
130 135 140
Gln Asp Met Leu Gln Phe Ile Lys Tyr Lys Asn Pro Glu Asp Glu Lys
145 150 155 160
Ile Lys Asn Tyr Ile Asp Asp Thr Leu Lys Gly Phe Phe Thr Tyr Phe
165 170 175
Gly Gly Phe Asn Gln Asn Arg Ala Asn Tyr Tyr Glu Thr Lys Lys Glu
180 185 190
Ala Ser Thr Ala Val Ala Thr Arg Ile Val His Glu Asn Leu Pro Lys
195 200 205
Phe Cys Asp Asn Val Ile Gln Phe Lys His Ile Ile Lys Arg Lys Lys
210 215 220
Asp Gly Thr Val Glu Lys Thr Glu Arg Lys Thr Glu Tyr Leu Asn Ala
225 230 235 240
Tyr Gln Tyr Leu Lys Asn Asn Asn Lys Ile Thr Gln Ile Lys Asp Ala
245 250 255
Glu Thr Glu Lys Met Ile Glu Ser Thr Pro Ile Ala Glu Lys Ile Phe
260 265 270
Asp Val Tyr Tyr Phe Ser Ser Cys Leu Ser Gln Lys Gln Ile Glu Glu
275 280 285
Tyr Asn Arg Ile Ile Gly His Tyr Asn Leu Leu Ile Asn Leu Tyr Asn
290 295 300
Gln Ala Lys Arg Ser Glu Gly Lys His Leu Ser Ala Asn Glu Lys Lys
305 310 315 320
Tyr Lys Asp Leu Pro Lys Phe Lys Thr Leu Tyr Lys Gln Ile Gly Cys
325 330 335
Gly Lys Lys Lys Asp Leu Phe Tyr Thr Ile Lys Cys Asp Thr Glu Glu
340 345 350
Glu Ala Asn Lys Ser Arg Asn Glu Gly Lys Glu Ser His Ser Val Glu
355 360 365
Glu Ile Ile Asn Lys Ala Gln Glu Ala Ile Asn Lys Tyr Phe Lys Ser
370 375 380
Asn Asn Asp Cys Glu Asn Ile Asn Thr Val Pro Asp Phe Ile Asn Tyr
385 390 395 400
Ile Leu Thr Lys Glu Asn Tyr Glu Gly Val Tyr Trp Ser Lys Ala Ala
405 410 415
Met Asn Thr Ile Ser Asp Lys Tyr Phe Ala Asn Tyr His Asp Leu Gln
420 425 430
Asp Arg Leu Lys Glu Ala Lys Val Phe Gln Lys Ala Asp Lys Lys Ser
435 440 445
Glu Asp Asp Ile Lys Ile Pro Glu Ala Ile Glu Leu Ser Gly Leu Phe
450 455 460
Gly Val Leu Asp Ser Leu Ala Asp Trp Gln Thr Thr Leu Phe Lys Ser
465 470 475 480
Ser Ile Leu Ser Asn Glu Asp Lys Leu Lys Ile Ile Thr Asp Ser Gln
485 490 495
Thr Pro Ser Glu Ala Leu Leu Lys Met Ile Phe Asn Asp Ile Glu Lys
500 505 510
Asn Met Glu Ser Phe Leu Lys Glu Thr Asn Asp Ile Ile Thr Leu Lys
515 520 525
Lys Tyr Lys Gly Asn Lys Glu Gly Thr Glu Lys Ile Lys Gln Trp Phe
530 535 540
Asp Tyr Thr Leu Ala Ile Asn Arg Met Leu Lys Tyr Phe Leu Val Lys
545 550 555 560
Glu Asn Lys Ile Lys Gly Asn Ser Leu Asp Thr Asn Ile Ser Glu Ala
565 570 575
Leu Lys Thr Leu Ile Tyr Ser Asp Asp Ala Glu Trp Phe Lys Trp Tyr
580 585 590
Asp Ala Leu Arg Asn Tyr Leu Thr Gln Lys Pro Gln Asp Glu Ala Lys
595 600 605
Glu Asn Lys Leu Lys Leu Asn Phe Asp Asn Pro Ser Leu Ala Gly Gly
610 615 620
Trp Asp Val Asn Lys Glu Cys Ser Asn Phe Cys Val Ile Leu Lys Asp
625 630 635 640
Lys Asn Glu Lys Lys Tyr Leu Ala Ile Met Lys Lys Gly Glu Asn Thr
645 650 655
Leu Phe Gln Lys Glu Trp Thr Glu Gly Arg Gly Lys Asn Leu Thr Lys
660 665 670
Lys Ser Asn Pro Leu Phe Glu Ile Asn Asn Cys Glu Ile Leu Ser Lys
675 680 685
Met Glu Tyr Asp Phe Trp Ala Asp Val Ser Lys Met Ile Pro Lys Cys
690 695 700
Ser Thr Gln Leu Lys Ala Val Val Asn His Phe Lys Gln Ser Asp Asn
705 710 715 720
Glu Phe Ile Phe Pro Ile Gly Tyr Lys Val Thr Ser Gly Glu Lys Phe
725 730 735
Arg Glu Glu Cys Lys Ile Ser Lys Gln Asp Phe Glu Leu Asn Asn Lys
740 745 750
Val Phe Asn Lys Asn Glu Leu Ser Val Thr Ala Met Arg Tyr Asp Leu
755 760 765
Ser Ser Thr Gln Glu Lys Gln Tyr Ile Lys Ala Phe Gln Lys Glu Tyr
770 775 780
Trp Glu Leu Leu Phe Lys Gln Glu Lys Arg Asp Thr Lys Leu Thr Asn
785 790 795 800
Asn Glu Ile Phe Asn Glu Trp Ile Asn Phe Cys Asn Lys Lys Tyr Ser
805 810 815
Glu Leu Leu Ser Trp Glu Arg Lys Tyr Lys Asp Ala Leu Thr Asn Trp
820 825 830
Ile Asn Phe Cys Lys Tyr Phe Leu Ser Lys Tyr Pro Lys Thr Thr Leu
835 840 845
Phe Asn Tyr Ser Phe Lys Glu Ser Glu Asn Tyr Asn Ser Leu Asp Glu
850 855 860
Phe Tyr Arg Asp Val Asp Ile Cys Ser Tyr Lys Leu Asn Ile Asn Thr
865 870 875 880
Thr Ile Asn Lys Ser Ile Leu Asp Arg Leu Val Glu Glu Gly Lys Leu
885 890 895
Tyr Leu Phe Glu Ile Lys Asn Gln Asp Ser Asn Asp Gly Lys Ser Ile
900 905 910
Gly His Lys Asn Asn Leu His Thr Ile Tyr Trp Asn Ala Ile Phe Glu
915 920 925
Asn Phe Asp Asn Arg Pro Lys Leu Asn Gly Glu Ala Glu Ile Phe Tyr
930 935 940
Arg Lys Ala Ile Ser Lys Asp Lys Leu Gly Ile Val Lys Gly Lys Lys
945 950 955 960
Thr Lys Asn Gly Thr Glu Ile Ile Lys Asn Tyr Arg Phe Ser Lys Glu
965 970 975
Lys Phe Ile Leu His Val Pro Ile Thr Leu Asn Phe Cys Ser Asn Asn
980 985 990
Glu Tyr Val Asn Asp Ile Val Asn Thr Lys Phe Tyr Asn Phe Ser Asn
995 1000 1005
Leu His Phe Leu Gly Ile Asp Arg Gly Glu Lys His Leu Ala Tyr
1010 1015 1020
Tyr Ser Leu Val Asn Lys Asn Gly Glu Ile Val Asp Gln Gly Thr
1025 1030 1035
Leu Asn Leu Pro Phe Thr Asp Lys Asp Gly Asn Gln Arg Ser Ile
1040 1045 1050
Lys Lys Glu Lys Tyr Phe Tyr Asn Lys Gln Glu Asp Lys Trp Glu
1055 1060 1065
Ala Lys Glu Val Asp Cys Trp Asn Tyr Asn Asp Leu Leu Asp Ala
1070 1075 1080
Met Ala Ser Asn Arg Asp Met Ala Arg Lys Asn Trp Gln Arg Ile
1085 1090 1095
Gly Thr Ile Lys Glu Ala Lys Asn Gly Tyr Val Ser Leu Val Ile
1100 1105 1110
Arg Lys Ile Ala Asp Leu Ala Val Asn Asn Glu Arg Pro Ala Phe
1115 1120 1125
Ile Val Leu Glu Asp Leu Asn Thr Gly Phe Lys Arg Ser Arg Gln
1130 1135 1140
Lys Ile Asp Lys Ser Val Tyr Gln Lys Phe Glu Leu Ala Leu Ala
1145 1150 1155
Lys Lys Leu Asn Phe Leu Val Asp Lys Asn Ala Lys Arg Asp Glu
1160 1165 1170
Ile Gly Ser Pro Thr Lys Ala Leu Gln Leu Thr Pro Pro Val Asn
1175 1180 1185
Asn Tyr Gly Asp Ile Glu Asn Lys Lys Gln Ala Gly Ile Met Leu
1190 1195 1200
Tyr Thr Arg Ala Asn Tyr Thr Ser Gln Thr Asp Pro Ala Thr Gly
1205 1210 1215
Trp Arg Lys Thr Ile Tyr Leu Lys Ala Gly Pro Glu Glu Thr Thr
1220 1225 1230
Tyr Lys Lys Asp Gly Lys Ile Lys Asn Lys Ser Val Lys Asp Gln
1235 1240 1245
Ile Ile Glu Thr Phe Thr Asp Ile Gly Phe Asp Gly Lys Asp Tyr
1250 1255 1260
Tyr Phe Glu Tyr Asp Lys Gly Glu Phe Val Asp Glu Lys Thr Gly
1265 1270 1275
Glu Ile Lys Pro Lys Lys Trp Arg Leu Tyr Ser Gly Glu Asn Gly
1280 1285 1290
Lys Ser Leu Asp Arg Phe Arg Gly Glu Arg Glu Lys Asp Lys Tyr
1295 1300 1305
Glu Trp Lys Ile Asp Lys Ile Asp Ile Val Lys Ile Leu Asp Asp
1310 1315 1320
Leu Phe Val Asn Phe Asp Lys Asn Ile Ser Leu Leu Lys Gln Leu
1325 1330 1335
Lys Glu Gly Val Glu Leu Thr Arg Asn Asn Glu His Gly Thr Gly
1340 1345 1350
Glu Ser Leu Arg Phe Ala Ile Asn Leu Ile Gln Gln Ile Arg Asn
1355 1360 1365
Thr Gly Asn Asn Glu Arg Asp Asn Asp Phe Ile Leu Ser Pro Val
1370 1375 1380
Arg Asp Glu Asn Gly Lys His Phe Asp Ser Arg Glu Tyr Trp Asp
1385 1390 1395
Lys Glu Thr Lys Gly Glu Lys Ile Ser Met Pro Ser Ser Gly Asp
1400 1405 1410
Ala Asn Gly Ala Phe Asn Ile Ala Arg Lys Gly Ile Ile Met Asn
1415 1420 1425
Ala His Ile Leu Ala Asn Ser Asp Ser Lys Asp Leu Ser Leu Phe
1430 1435 1440
Val Ser Asp Glu Glu Trp Asp Leu His Leu Asn Asn Lys Thr Glu
1445 1450 1455
Trp Lys Lys Gln Leu Asn Ile Phe Ser Ser Arg Lys Ala Met Ala
1460 1465 1470
Lys Arg Lys Lys
1475
<210> 33
<211> 1285
<212> PRT
<213> 罗伊兹曼菌门细菌暂定种
<220>
<221> MISC_FEATURE
<222> (1)..(1285)
<223> Genbank KKQ38174 Cpf1
<400> 33
Met Lys Ser Phe Asp Ser Phe Thr Asn Leu Tyr Ser Leu Ser Lys Thr
1 5 10 15
Leu Lys Phe Glu Met Arg Pro Val Gly Asn Thr Gln Lys Met Leu Asp
20 25 30
Asn Ala Gly Val Phe Glu Lys Asp Lys Leu Ile Gln Lys Lys Tyr Gly
35 40 45
Lys Thr Lys Pro Tyr Phe Asp Arg Leu His Arg Glu Phe Ile Glu Glu
50 55 60
Ala Leu Thr Gly Val Glu Leu Ile Gly Leu Asp Glu Asn Phe Arg Thr
65 70 75 80
Leu Val Asp Trp Gln Lys Asp Lys Lys Asn Asn Val Ala Met Lys Ala
85 90 95
Tyr Glu Asn Ser Leu Gln Arg Leu Arg Thr Glu Ile Gly Lys Ile Phe
100 105 110
Asn Leu Lys Ala Glu Asp Trp Val Lys Asn Lys Tyr Pro Ile Leu Gly
115 120 125
Leu Lys Asn Lys Asn Thr Asp Ile Leu Phe Glu Glu Ala Val Phe Gly
130 135 140
Ile Leu Lys Ala Arg Tyr Gly Glu Glu Lys Asp Thr Phe Ile Glu Val
145 150 155 160
Glu Glu Ile Asp Lys Thr Gly Lys Ser Lys Ile Asn Gln Ile Ser Ile
165 170 175
Phe Asp Ser Trp Lys Gly Phe Thr Gly Tyr Phe Lys Lys Phe Phe Glu
180 185 190
Thr Arg Lys Asn Phe Tyr Lys Asn Asp Gly Thr Ser Thr Ala Ile Ala
195 200 205
Thr Arg Ile Ile Asp Gln Asn Leu Lys Arg Phe Ile Asp Asn Leu Ser
210 215 220
Ile Val Glu Ser Val Arg Gln Lys Val Asp Leu Ala Glu Thr Glu Lys
225 230 235 240
Ser Phe Ser Ile Ser Leu Ser Gln Phe Phe Ser Ile Asp Phe Tyr Asn
245 250 255
Lys Cys Leu Leu Gln Asp Gly Ile Asp Tyr Tyr Asn Lys Ile Ile Gly
260 265 270
Gly Glu Thr Leu Lys Asn Gly Glu Lys Leu Ile Gly Leu Asn Glu Leu
275 280 285
Ile Asn Gln Tyr Arg Gln Asn Asn Lys Asp Gln Lys Ile Pro Phe Phe
290 295 300
Lys Leu Leu Asp Lys Gln Ile Leu Ser Glu Lys Ile Leu Phe Leu Asp
305 310 315 320
Glu Ile Lys Asn Asp Thr Glu Leu Ile Glu Ala Leu Ser Gln Phe Ala
325 330 335
Lys Thr Ala Glu Glu Lys Thr Lys Ile Val Lys Lys Leu Phe Ala Asp
340 345 350
Phe Val Glu Asn Asn Ser Lys Tyr Asp Leu Ala Gln Ile Tyr Ile Ser
355 360 365
Gln Glu Ala Phe Asn Thr Ile Ser Asn Lys Trp Thr Ser Glu Thr Glu
370 375 380
Thr Phe Ala Lys Tyr Leu Phe Glu Ala Met Lys Ser Gly Lys Leu Ala
385 390 395 400
Lys Tyr Glu Lys Lys Asp Asn Ser Tyr Lys Phe Pro Asp Phe Ile Ala
405 410 415
Leu Ser Gln Met Lys Ser Ala Leu Leu Ser Ile Ser Leu Glu Gly His
420 425 430
Phe Trp Lys Glu Lys Tyr Tyr Lys Ile Ser Lys Phe Gln Glu Lys Thr
435 440 445
Asn Trp Glu Gln Phe Leu Ala Ile Phe Leu Tyr Glu Phe Asn Ser Leu
450 455 460
Phe Ser Asp Lys Ile Asn Thr Lys Asp Gly Glu Thr Lys Gln Val Gly
465 470 475 480
Tyr Tyr Leu Phe Ala Lys Asp Leu His Asn Leu Ile Leu Ser Glu Gln
485 490 495
Ile Asp Ile Pro Lys Asp Ser Lys Val Thr Ile Lys Asp Phe Ala Asp
500 505 510
Ser Val Leu Thr Ile Tyr Gln Met Ala Lys Tyr Phe Ala Val Glu Lys
515 520 525
Lys Arg Ala Trp Leu Ala Glu Tyr Glu Leu Asp Ser Phe Tyr Thr Gln
530 535 540
Pro Asp Thr Gly Tyr Leu Gln Phe Tyr Asp Asn Ala Tyr Glu Asp Ile
545 550 555 560
Val Gln Val Tyr Asn Lys Leu Arg Asn Tyr Leu Thr Lys Lys Pro Tyr
565 570 575
Ser Glu Glu Lys Trp Lys Leu Asn Phe Glu Asn Ser Thr Leu Ala Asn
580 585 590
Gly Trp Asp Lys Asn Lys Glu Ser Asp Asn Ser Ala Val Ile Leu Gln
595 600 605
Lys Gly Gly Lys Tyr Tyr Leu Gly Leu Ile Thr Lys Gly His Asn Lys
610 615 620
Ile Phe Asp Asp Arg Phe Gln Glu Lys Phe Ile Val Gly Ile Glu Gly
625 630 635 640
Gly Lys Tyr Glu Lys Ile Val Tyr Lys Phe Phe Pro Asp Gln Ala Lys
645 650 655
Met Phe Pro Lys Val Cys Phe Ser Ala Lys Gly Leu Glu Phe Phe Arg
660 665 670
Pro Ser Glu Glu Ile Leu Arg Ile Tyr Asn Asn Ala Glu Phe Lys Lys
675 680 685
Gly Glu Thr Tyr Ser Ile Asp Ser Met Gln Lys Leu Ile Asp Phe Tyr
690 695 700
Lys Asp Cys Leu Thr Lys Tyr Glu Gly Trp Ala Cys Tyr Thr Phe Arg
705 710 715 720
His Leu Lys Pro Thr Glu Glu Tyr Gln Asn Asn Ile Gly Glu Phe Phe
725 730 735
Arg Asp Val Ala Glu Asp Gly Tyr Arg Ile Asp Phe Gln Gly Ile Ser
740 745 750
Asp Gln Tyr Ile His Glu Lys Asn Glu Lys Gly Glu Leu His Leu Phe
755 760 765
Glu Ile His Asn Lys Asp Trp Asn Leu Asp Lys Ala Arg Asp Gly Lys
770 775 780
Ser Lys Thr Thr Gln Lys Asn Leu His Thr Leu Tyr Phe Glu Ser Leu
785 790 795 800
Phe Ser Asn Asp Asn Val Val Gln Asn Phe Pro Ile Lys Leu Asn Gly
805 810 815
Gln Ala Glu Ile Phe Tyr Arg Pro Lys Thr Glu Lys Asp Lys Leu Glu
820 825 830
Ser Lys Lys Asp Lys Lys Gly Asn Lys Val Ile Asp His Lys Arg Tyr
835 840 845
Ser Glu Asn Lys Ile Phe Phe His Val Pro Leu Thr Leu Asn Arg Thr
850 855 860
Lys Asn Asp Ser Tyr Arg Phe Asn Ala Gln Ile Asn Asn Phe Leu Ala
865 870 875 880
Asn Asn Lys Asp Ile Asn Ile Ile Gly Val Asp Arg Gly Glu Lys His
885 890 895
Leu Val Tyr Tyr Ser Val Ile Thr Gln Ala Ser Asp Ile Leu Glu Ser
900 905 910
Gly Ser Leu Asn Glu Leu Asn Gly Val Asn Tyr Ala Glu Lys Leu Gly
915 920 925
Lys Lys Ala Glu Asn Arg Glu Gln Ala Arg Arg Asp Trp Gln Asp Val
930 935 940
Gln Gly Ile Lys Asp Leu Lys Lys Gly Tyr Ile Ser Gln Val Val Arg
945 950 955 960
Lys Leu Ala Asp Leu Ala Ile Lys His Asn Ala Ile Ile Ile Leu Glu
965 970 975
Asp Leu Asn Met Arg Phe Lys Gln Val Arg Gly Gly Ile Glu Lys Ser
980 985 990
Ile Tyr Gln Gln Leu Glu Lys Ala Leu Ile Asp Lys Leu Ser Phe Leu
995 1000 1005
Val Asp Lys Gly Glu Lys Asn Pro Glu Gln Ala Gly His Leu Leu
1010 1015 1020
Lys Ala Tyr Gln Leu Ser Ala Pro Phe Glu Thr Phe Gln Lys Met
1025 1030 1035
Gly Lys Gln Thr Gly Ile Ile Phe Tyr Thr Gln Ala Ser Tyr Thr
1040 1045 1050
Ser Lys Ser Asp Pro Val Thr Gly Trp Arg Pro His Leu Tyr Leu
1055 1060 1065
Lys Tyr Phe Ser Ala Lys Lys Ala Lys Asp Asp Ile Ala Lys Phe
1070 1075 1080
Thr Lys Ile Glu Phe Val Asn Asp Arg Phe Glu Leu Thr Tyr Asp
1085 1090 1095
Ile Lys Asp Phe Gln Gln Ala Lys Glu Tyr Pro Asn Lys Thr Val
1100 1105 1110
Trp Lys Val Cys Ser Asn Val Glu Arg Phe Arg Trp Asp Lys Asn
1115 1120 1125
Leu Asn Gln Asn Lys Gly Gly Tyr Thr His Tyr Thr Asn Ile Thr
1130 1135 1140
Glu Asn Ile Gln Glu Leu Phe Thr Lys Tyr Gly Ile Asp Ile Thr
1145 1150 1155
Lys Asp Leu Leu Thr Gln Ile Ser Thr Ile Asp Glu Lys Gln Asn
1160 1165 1170
Thr Ser Phe Phe Arg Asp Phe Ile Phe Tyr Phe Asn Leu Ile Cys
1175 1180 1185
Gln Ile Arg Asn Thr Asp Asp Ser Glu Ile Ala Lys Lys Asn Gly
1190 1195 1200
Lys Asp Asp Phe Ile Leu Ser Pro Val Glu Pro Phe Phe Asp Ser
1205 1210 1215
Arg Lys Asp Asn Gly Asn Lys Leu Pro Glu Asn Gly Asp Asp Asn
1220 1225 1230
Gly Ala Tyr Asn Ile Ala Arg Lys Gly Ile Val Ile Leu Asn Lys
1235 1240 1245
Ile Ser Gln Tyr Ser Glu Lys Asn Glu Asn Cys Glu Lys Met Lys
1250 1255 1260
Trp Gly Asp Leu Tyr Val Ser Asn Ile Asp Trp Asp Asn Phe Val
1265 1270 1275
Thr Gln Ala Asn Ala Arg His
1280 1285
<210> 34
<211> 1403
<212> PRT
<213> 法尔科夫菌门细菌暂定种
<220>
<221> MISC_FEATURE
<222> (1)..(1403)
<223> Genbank KKR91555 Cpf1
<400> 34
Met Leu Phe Phe Met Ser Thr Asp Ile Thr Asn Lys Pro Arg Glu Lys
1 5 10 15
Gly Val Phe Asp Asn Phe Thr Asn Leu Tyr Glu Phe Ser Lys Thr Leu
20 25 30
Thr Phe Gly Leu Ile Pro Leu Lys Trp Asp Asp Asn Lys Lys Met Ile
35 40 45
Val Glu Asp Glu Asp Phe Ser Val Leu Arg Lys Tyr Gly Val Ile Glu
50 55 60
Glu Asp Lys Arg Ile Ala Glu Ser Ile Lys Ile Ala Lys Phe Tyr Leu
65 70 75 80
Asn Ile Leu His Arg Glu Leu Ile Gly Lys Val Leu Gly Ser Leu Lys
85 90 95
Phe Glu Lys Lys Asn Leu Glu Asn Tyr Asp Arg Leu Leu Gly Glu Ile
100 105 110
Glu Lys Asn Asn Lys Asn Glu Asn Ile Ser Glu Asp Lys Lys Lys Glu
115 120 125
Ile Arg Lys Asn Phe Lys Lys Glu Leu Ser Ile Ala Gln Asp Ile Leu
130 135 140
Leu Lys Lys Val Gly Glu Val Phe Glu Ser Asn Gly Ser Gly Ile Leu
145 150 155 160
Ser Ser Lys Asn Cys Leu Asp Glu Leu Thr Lys Arg Phe Thr Arg Gln
165 170 175
Glu Val Asp Lys Leu Arg Arg Glu Asn Lys Asp Ile Gly Val Glu Tyr
180 185 190
Pro Asp Val Ala Tyr Arg Glu Lys Asp Gly Lys Glu Glu Thr Lys Ser
195 200 205
Phe Phe Ala Met Asp Val Gly Tyr Leu Asp Asp Phe His Lys Asn Arg
210 215 220
Lys Gln Leu Tyr Ser Val Lys Gly Lys Lys Asn Ser Leu Gly Arg Arg
225 230 235 240
Ile Leu Asp Asn Phe Glu Ile Phe Cys Lys Asn Lys Lys Leu Tyr Glu
245 250 255
Lys Tyr Lys Asn Leu Asp Ile Asp Phe Ser Glu Ile Glu Arg Asn Phe
260 265 270
Asn Leu Thr Leu Glu Lys Val Phe Asp Phe Asp Asn Tyr Asn Glu Arg
275 280 285
Leu Thr Gln Glu Gly Leu Asp Glu Tyr Ala Lys Ile Leu Gly Gly Glu
290 295 300
Ser Asn Lys Gln Glu Arg Thr Ala Asn Ile His Gly Leu Asn Gln Ile
305 310 315 320
Ile Asn Leu Tyr Ile Gln Lys Lys Gln Ser Glu Gln Lys Ala Glu Gln
325 330 335
Lys Glu Thr Gly Lys Lys Lys Ile Lys Phe Asn Lys Lys Asp Tyr Pro
340 345 350
Thr Phe Thr Cys Leu Gln Lys Gln Ile Leu Ser Gln Val Phe Arg Lys
355 360 365
Glu Ile Ile Ile Glu Ser Asp Arg Asp Leu Ile Arg Glu Leu Lys Phe
370 375 380
Phe Val Glu Glu Ser Lys Glu Lys Val Asp Lys Ala Arg Gly Ile Ile
385 390 395 400
Glu Phe Leu Leu Asn His Glu Glu Asn Asp Ile Asp Leu Ala Met Val
405 410 415
Tyr Leu Pro Lys Ser Lys Ile Asn Ser Phe Val Tyr Lys Val Phe Lys
420 425 430
Glu Pro Gln Asp Phe Leu Ser Val Phe Gln Asp Gly Ala Ser Asn Leu
435 440 445
Asp Phe Val Ser Phe Asp Lys Ile Lys Thr His Leu Glu Asn Asn Lys
450 455 460
Leu Thr Tyr Lys Ile Phe Phe Lys Thr Leu Ile Lys Glu Asn His Asp
465 470 475 480
Phe Glu Ser Phe Leu Ile Leu Leu Gln Gln Glu Ile Asp Leu Leu Ile
485 490 495
Asp Gly Gly Glu Thr Val Thr Leu Gly Gly Lys Lys Glu Ser Ile Thr
500 505 510
Ser Leu Asp Glu Lys Lys Asn Arg Leu Lys Glu Lys Leu Gly Trp Phe
515 520 525
Glu Gly Lys Val Arg Glu Asn Glu Lys Met Lys Asp Glu Glu Glu Gly
530 535 540
Glu Phe Cys Ser Thr Val Leu Ala Tyr Ser Gln Ala Val Leu Asn Ile
545 550 555 560
Thr Lys Arg Ala Glu Ile Phe Trp Leu Asn Glu Lys Gln Asp Ala Lys
565 570 575
Val Gly Glu Asp Asn Lys Asp Met Ile Phe Tyr Lys Lys Phe Asp Glu
580 585 590
Phe Ala Asp Asp Gly Phe Ala Pro Phe Phe Tyr Phe Asp Lys Phe Gly
595 600 605
Asn Tyr Leu Lys Arg Arg Ser Arg Asn Thr Thr Lys Glu Ile Lys Leu
610 615 620
His Phe Gly Asn Asp Asp Leu Leu Glu Gly Trp Asp Met Asn Lys Glu
625 630 635 640
Pro Glu Tyr Trp Ser Phe Ile Leu Arg Asp Arg Asn Gln Tyr Tyr Leu
645 650 655
Gly Ile Gly Lys Lys Asp Gly Glu Ile Phe His Lys Lys Leu Gly Asn
660 665 670
Ser Val Glu Ala Val Lys Glu Ala Tyr Glu Leu Glu Asn Glu Ala Asp
675 680 685
Phe Tyr Glu Lys Ile Asp Tyr Lys Gln Leu Asn Ile Asp Arg Phe Glu
690 695 700
Gly Ile Ala Phe Pro Lys Lys Thr Lys Thr Glu Glu Ala Phe Arg Gln
705 710 715 720
Val Cys Lys Lys Arg Ala Asp Glu Phe Leu Gly Gly Asp Thr Tyr Glu
725 730 735
Phe Lys Ile Leu Leu Ala Ile Lys Lys Glu Tyr Asp Asp Phe Lys Ala
740 745 750
Arg Arg Gln Lys Glu Lys Asp Trp Asp Ser Lys Phe Ser Lys Glu Lys
755 760 765
Met Ser Lys Leu Ile Glu Tyr Tyr Ile Thr Cys Leu Gly Lys Arg Asp
770 775 780
Asp Trp Lys Arg Phe Asn Leu Asn Phe Arg Gln Pro Lys Glu Tyr Glu
785 790 795 800
Asp Arg Ser Asp Phe Val Arg His Ile Gln Arg Gln Ala Tyr Trp Ile
805 810 815
Asp Pro Arg Lys Val Ser Lys Asp Tyr Val Asp Lys Lys Val Ala Glu
820 825 830
Gly Glu Met Phe Leu Phe Lys Val His Asn Lys Asp Phe Tyr Asp Phe
835 840 845
Glu Arg Lys Ser Glu Asp Lys Lys Asn His Thr Ala Asn Leu Phe Thr
850 855 860
Gln Tyr Leu Leu Glu Leu Phe Ser Cys Glu Asn Ile Lys Asn Ile Lys
865 870 875 880
Ser Lys Asp Leu Ile Glu Ser Ile Phe Glu Leu Asp Gly Lys Ala Glu
885 890 895
Ile Arg Phe Arg Pro Lys Thr Asp Asp Val Lys Leu Lys Ile Tyr Gln
900 905 910
Lys Lys Gly Lys Asp Val Thr Tyr Ala Asp Lys Arg Asp Gly Asn Lys
915 920 925
Glu Lys Glu Val Ile Gln His Arg Arg Phe Ala Lys Asp Ala Leu Thr
930 935 940
Leu His Leu Lys Ile Arg Leu Asn Phe Gly Lys His Val Asn Leu Phe
945 950 955 960
Asp Phe Asn Lys Leu Val Asn Thr Glu Leu Phe Ala Lys Val Pro Val
965 970 975
Lys Ile Leu Gly Met Asp Arg Gly Glu Asn Asn Leu Ile Tyr Tyr Cys
980 985 990
Phe Leu Asp Glu His Gly Glu Ile Glu Asn Gly Lys Cys Gly Ser Leu
995 1000 1005
Asn Arg Val Gly Glu Gln Ile Ile Thr Leu Glu Asp Asp Lys Lys
1010 1015 1020
Val Lys Glu Pro Val Asp Tyr Phe Gln Leu Leu Val Asp Arg Glu
1025 1030 1035
Gly Gln Arg Asp Trp Glu Gln Lys Asn Trp Gln Lys Met Thr Arg
1040 1045 1050
Ile Lys Asp Leu Lys Lys Ala Tyr Leu Gly Asn Val Val Ser Trp
1055 1060 1065
Ile Ser Lys Glu Met Leu Ser Gly Ile Lys Glu Gly Val Val Thr
1070 1075 1080
Ile Gly Val Leu Glu Asp Leu Asn Ser Asn Phe Lys Arg Thr Arg
1085 1090 1095
Phe Phe Arg Glu Arg Gln Val Tyr Gln Gly Phe Glu Lys Ala Leu
1100 1105 1110
Val Asn Lys Leu Gly Tyr Leu Val Asp Lys Lys Tyr Asp Asn Tyr
1115 1120 1125
Arg Asn Val Tyr Gln Phe Ala Pro Ile Val Asp Ser Val Glu Glu
1130 1135 1140
Met Glu Lys Asn Lys Gln Ile Gly Thr Leu Val Tyr Val Pro Ala
1145 1150 1155
Ser Tyr Thr Ser Lys Ile Cys Pro His Pro Lys Cys Gly Trp Arg
1160 1165 1170
Glu Arg Leu Tyr Met Lys Asn Ser Ala Ser Lys Glu Lys Ile Val
1175 1180 1185
Gly Leu Leu Lys Ser Asp Gly Ile Lys Ile Ser Tyr Asp Gln Lys
1190 1195 1200
Asn Asp Arg Phe Tyr Phe Glu Tyr Gln Trp Glu Gln Glu His Lys
1205 1210 1215
Ser Asp Gly Lys Lys Lys Lys Tyr Ser Gly Val Asp Lys Val Phe
1220 1225 1230
Ser Asn Val Ser Arg Met Arg Trp Asp Val Glu Gln Lys Lys Ser
1235 1240 1245
Ile Asp Phe Val Asp Gly Thr Asp Gly Ser Ile Thr Asn Lys Leu
1250 1255 1260
Lys Ser Leu Leu Lys Gly Lys Gly Ile Glu Leu Asp Asn Ile Asn
1265 1270 1275
Gln Gln Ile Val Asn Gln Gln Lys Glu Leu Gly Val Glu Phe Phe
1280 1285 1290
Gln Ser Ile Ile Phe Tyr Phe Asn Leu Ile Met Gln Ile Arg Asn
1295 1300 1305
Tyr Asp Lys Glu Lys Ser Gly Ser Glu Ala Asp Tyr Ile Gln Cys
1310 1315 1320
Pro Ser Cys Leu Phe Asp Ser Arg Lys Pro Glu Met Asn Gly Lys
1325 1330 1335
Leu Ser Ala Ile Thr Asn Gly Asp Ala Asn Gly Ala Tyr Asn Ile
1340 1345 1350
Ala Arg Lys Gly Phe Met Gln Leu Cys Arg Ile Arg Glu Asn Pro
1355 1360 1365
Gln Glu Pro Met Lys Leu Ile Thr Asn Arg Glu Trp Asp Glu Ala
1370 1375 1380
Val Arg Glu Trp Asp Ile Tyr Ser Ala Ala Gln Lys Ile Pro Val
1385 1390 1395
Leu Ser Glu Glu Asn
1400
<210> 35
<211> 1352
<212> PRT
<213> 俭菌总门细菌群
<220>
<221> MISC_FEATURE
<222> (1)..(1352)
<223> Genbank KKT48220 Cpf1
<400> 35
Met Glu Asn Ile Phe Asp Gln Phe Ile Gly Lys Tyr Ser Leu Ser Lys
1 5 10 15
Thr Leu Arg Phe Glu Leu Lys Pro Val Gly Lys Thr Glu Asp Phe Leu
20 25 30
Lys Ile Asn Lys Val Phe Glu Lys Asp Gln Thr Ile Asp Asp Ser Tyr
35 40 45
Asn Gln Ala Lys Phe Tyr Phe Asp Ser Leu His Gln Lys Phe Ile Asp
50 55 60
Ala Ala Leu Ala Ser Asp Lys Thr Ser Glu Leu Ser Phe Gln Asn Phe
65 70 75 80
Ala Asp Val Leu Glu Lys Gln Asn Lys Ile Ile Leu Asp Lys Lys Arg
85 90 95
Glu Met Gly Ala Leu Arg Lys Arg Asp Lys Asn Ala Val Gly Ile Asp
100 105 110
Arg Leu Gln Lys Glu Ile Asn Asp Ala Glu Asp Ile Ile Gln Lys Glu
115 120 125
Lys Glu Lys Ile Tyr Lys Asp Val Arg Thr Leu Phe Asp Asn Glu Ala
130 135 140
Glu Ser Trp Lys Thr Tyr Tyr Gln Glu Arg Glu Val Asp Gly Lys Lys
145 150 155 160
Ile Thr Phe Ser Lys Ala Asp Leu Lys Gln Lys Gly Ala Asp Phe Leu
165 170 175
Thr Ala Ala Gly Ile Leu Lys Val Leu Lys Tyr Glu Phe Pro Glu Glu
180 185 190
Lys Glu Lys Glu Phe Gln Ala Lys Asn Gln Pro Ser Leu Phe Val Glu
195 200 205
Glu Lys Glu Asn Pro Gly Gln Lys Arg Tyr Ile Phe Asp Ser Phe Asp
210 215 220
Lys Phe Ala Gly Tyr Leu Thr Lys Phe Gln Gln Thr Lys Lys Asn Leu
225 230 235 240
Tyr Ala Ala Asp Gly Thr Ser Thr Ala Val Ala Thr Arg Ile Ala Asp
245 250 255
Asn Phe Ile Ile Phe His Gln Asn Thr Lys Val Phe Arg Asp Lys Tyr
260 265 270
Lys Asn Asn His Thr Asp Leu Gly Phe Asp Glu Glu Asn Ile Phe Glu
275 280 285
Ile Glu Arg Tyr Lys Asn Cys Leu Leu Gln Arg Glu Ile Glu His Ile
290 295 300
Lys Asn Glu Asn Ser Tyr Asn Lys Ile Ile Gly Arg Ile Asn Lys Lys
305 310 315 320
Ile Lys Glu Tyr Arg Asp Gln Lys Ala Lys Asp Thr Lys Leu Thr Lys
325 330 335
Ser Asp Phe Pro Phe Phe Lys Asn Leu Asp Lys Gln Ile Leu Gly Glu
340 345 350
Val Glu Lys Glu Lys Gln Leu Ile Glu Lys Thr Arg Glu Lys Thr Glu
355 360 365
Glu Asp Val Leu Ile Glu Arg Phe Lys Glu Phe Ile Glu Asn Asn Glu
370 375 380
Glu Arg Phe Thr Ala Ala Lys Lys Leu Met Asn Ala Phe Cys Asn Gly
385 390 395 400
Glu Phe Glu Ser Glu Tyr Glu Gly Ile Tyr Leu Lys Asn Lys Ala Ile
405 410 415
Asn Thr Ile Ser Arg Arg Trp Phe Val Ser Asp Arg Asp Phe Glu Leu
420 425 430
Lys Leu Pro Gln Gln Lys Ser Lys Asn Lys Ser Glu Lys Asn Glu Pro
435 440 445
Lys Val Lys Lys Phe Ile Ser Ile Ala Glu Ile Lys Asn Ala Val Glu
450 455 460
Glu Leu Asp Gly Asp Ile Phe Lys Ala Val Phe Tyr Asp Lys Lys Ile
465 470 475 480
Ile Ala Gln Gly Gly Ser Lys Leu Glu Gln Phe Leu Val Ile Trp Lys
485 490 495
Tyr Glu Phe Glu Tyr Leu Phe Arg Asp Ile Glu Arg Glu Asn Gly Glu
500 505 510
Lys Leu Leu Gly Tyr Asp Ser Cys Leu Lys Ile Ala Lys Gln Leu Gly
515 520 525
Ile Phe Pro Gln Glu Lys Glu Ala Arg Glu Lys Ala Thr Ala Val Ile
530 535 540
Lys Asn Tyr Ala Asp Ala Gly Leu Gly Ile Phe Gln Met Met Lys Tyr
545 550 555 560
Phe Ser Leu Asp Asp Lys Asp Arg Lys Asn Thr Pro Gly Gln Leu Ser
565 570 575
Thr Asn Phe Tyr Ala Glu Tyr Asp Gly Tyr Tyr Lys Asp Phe Glu Phe
580 585 590
Ile Lys Tyr Tyr Asn Glu Phe Arg Asn Phe Ile Thr Lys Lys Pro Phe
595 600 605
Asp Glu Asp Lys Ile Lys Leu Asn Phe Glu Asn Gly Ala Leu Leu Lys
610 615 620
Gly Trp Asp Glu Asn Lys Glu Tyr Asp Phe Met Gly Val Ile Leu Lys
625 630 635 640
Lys Glu Gly Arg Leu Tyr Leu Gly Ile Met His Lys Asn His Arg Lys
645 650 655
Leu Phe Gln Ser Met Gly Asn Ala Lys Gly Asp Asn Ala Asn Arg Tyr
660 665 670
Gln Lys Met Ile Tyr Lys Gln Ile Ala Asp Ala Ser Lys Asp Val Pro
675 680 685
Arg Leu Leu Leu Thr Ser Lys Lys Ala Met Glu Lys Phe Lys Pro Ser
690 695 700
Gln Glu Ile Leu Arg Ile Lys Lys Glu Lys Thr Phe Lys Arg Glu Ser
705 710 715 720
Lys Asn Phe Ser Leu Arg Asp Leu His Ala Leu Ile Glu Tyr Tyr Arg
725 730 735
Asn Cys Ile Pro Gln Tyr Ser Asn Trp Ser Phe Tyr Asp Phe Gln Phe
740 745 750
Gln Asp Thr Gly Lys Tyr Gln Asn Ile Lys Glu Phe Thr Asp Asp Val
755 760 765
Gln Lys Tyr Gly Tyr Lys Ile Ser Phe Arg Asp Ile Asp Asp Glu Tyr
770 775 780
Ile Asn Gln Ala Leu Asn Glu Gly Lys Met Tyr Leu Phe Glu Val Val
785 790 795 800
Asn Lys Asp Ile Tyr Asn Thr Lys Asn Gly Ser Lys Asn Leu His Thr
805 810 815
Leu Tyr Phe Glu His Ile Leu Ser Ala Glu Asn Leu Asn Asp Pro Val
820 825 830
Phe Lys Leu Ser Gly Met Ala Glu Ile Phe Gln Arg Gln Pro Ser Val
835 840 845
Asn Glu Arg Glu Lys Ile Thr Thr Gln Lys Asn Gln Cys Ile Leu Asp
850 855 860
Lys Gly Asp Arg Ala Tyr Lys Tyr Arg Arg Tyr Thr Glu Lys Lys Ile
865 870 875 880
Met Phe His Met Ser Leu Val Leu Asn Thr Gly Lys Gly Glu Ile Lys
885 890 895
Gln Val Gln Phe Asn Lys Ile Ile Asn Gln Arg Ile Ser Ser Ser Asp
900 905 910
Asn Glu Met Arg Val Asn Val Ile Gly Ile Asp Arg Gly Glu Lys Asn
915 920 925
Leu Leu Tyr Tyr Ser Val Val Lys Gln Asn Gly Glu Ile Ile Glu Gln
930 935 940
Ala Ser Leu Asn Glu Ile Asn Gly Val Asn Tyr Arg Asp Lys Leu Ile
945 950 955 960
Glu Arg Glu Lys Glu Arg Leu Lys Asn Arg Gln Ser Trp Lys Pro Val
965 970 975
Val Lys Ile Lys Asp Leu Lys Lys Gly Tyr Ile Ser His Val Ile His
980 985 990
Lys Ile Cys Gln Leu Ile Glu Lys Tyr Ser Ala Ile Val Val Leu Glu
995 1000 1005
Asp Leu Asn Met Arg Phe Lys Gln Ile Arg Gly Gly Ile Glu Arg
1010 1015 1020
Ser Val Tyr Gln Gln Phe Glu Lys Ala Leu Ile Asp Lys Leu Gly
1025 1030 1035
Tyr Leu Val Phe Lys Asp Asn Arg Asp Leu Arg Ala Pro Gly Gly
1040 1045 1050
Val Leu Asn Gly Tyr Gln Leu Ser Ala Pro Phe Val Ser Phe Glu
1055 1060 1065
Lys Met Arg Lys Gln Thr Gly Ile Leu Phe Tyr Thr Gln Ala Glu
1070 1075 1080
Tyr Thr Ser Lys Thr Asp Pro Ile Thr Gly Phe Arg Lys Asn Val
1085 1090 1095
Tyr Ile Ser Asn Ser Ala Ser Leu Asp Lys Ile Lys Glu Ala Val
1100 1105 1110
Lys Lys Phe Asp Ala Ile Gly Trp Asp Gly Lys Glu Gln Ser Tyr
1115 1120 1125
Phe Phe Lys Tyr Asn Pro Tyr Asn Leu Ala Asp Glu Lys Tyr Lys
1130 1135 1140
Asn Ser Thr Val Ser Lys Glu Trp Ala Ile Phe Ala Ser Ala Pro
1145 1150 1155
Arg Ile Arg Arg Gln Lys Gly Glu Asp Gly Tyr Trp Lys Tyr Asp
1160 1165 1170
Arg Val Lys Val Asn Glu Glu Phe Glu Lys Leu Leu Lys Val Trp
1175 1180 1185
Asn Phe Val Asn Pro Lys Ala Thr Asp Ile Lys Gln Glu Ile Ile
1190 1195 1200
Lys Lys Glu Lys Ala Gly Asp Leu Gln Gly Glu Lys Glu Leu Asp
1205 1210 1215
Gly Arg Leu Arg Asn Phe Trp His Ser Phe Ile Tyr Leu Phe Asn
1220 1225 1230
Leu Val Leu Glu Leu Arg Asn Ser Phe Ser Leu Gln Ile Lys Ile
1235 1240 1245
Lys Ala Gly Glu Val Ile Ala Val Asp Glu Gly Val Asp Phe Ile
1250 1255 1260
Ala Ser Pro Val Lys Pro Phe Phe Thr Thr Pro Asn Pro Tyr Ile
1265 1270 1275
Pro Ser Asn Leu Cys Trp Leu Ala Val Glu Asn Ala Asp Ala Asn
1280 1285 1290
Gly Ala Tyr Asn Ile Ala Arg Lys Gly Val Met Ile Leu Lys Lys
1295 1300 1305
Ile Arg Glu His Ala Lys Lys Asp Pro Glu Phe Lys Lys Leu Pro
1310 1315 1320
Asn Leu Phe Ile Ser Asn Ala Glu Trp Asp Glu Ala Ala Arg Asp
1325 1330 1335
Trp Gly Lys Tyr Ala Gly Thr Thr Ala Leu Asn Leu Asp His
1340 1345 1350
<210> 36
<211> 1298
<212> PRT
<213> 硫微螺菌
<220>
<221> MISC_FEATURE
<222> (1)..(1298)
<223> Genbank KUJ74576 Cpf1
<400> 36
Met Thr Lys Thr Phe Asp Ser Glu Phe Phe Asn Leu Tyr Ser Leu Gln
1 5 10 15
Lys Thr Val Arg Phe Glu Leu Lys Pro Val Gly Glu Thr Ala Ser Phe
20 25 30
Val Glu Asp Phe Lys Asn Glu Gly Leu Lys Arg Val Val Ser Glu Asp
35 40 45
Glu Arg Arg Ala Val Asp Tyr Gln Lys Val Lys Glu Ile Ile Asp Asp
50 55 60
Tyr His Arg Asp Phe Ile Glu Glu Ser Leu Asn Tyr Phe Pro Glu Gln
65 70 75 80
Val Ser Lys Asp Ala Leu Glu Gln Ala Phe His Leu Tyr Gln Lys Leu
85 90 95
Lys Ala Ala Lys Val Glu Glu Arg Glu Lys Ala Leu Lys Glu Trp Glu
100 105 110
Ala Leu Gln Lys Lys Leu Arg Glu Lys Val Val Lys Cys Phe Ser Asp
115 120 125
Ser Asn Lys Ala Arg Phe Ser Arg Ile Asp Lys Lys Glu Leu Ile Lys
130 135 140
Glu Asp Leu Ile Asn Trp Leu Val Ala Gln Asn Arg Glu Asp Asp Ile
145 150 155 160
Pro Thr Val Glu Thr Phe Asn Asn Phe Thr Thr Tyr Phe Thr Gly Phe
165 170 175
His Glu Asn Arg Lys Asn Ile Tyr Ser Lys Asp Asp His Ala Thr Ala
180 185 190
Ile Ser Phe Arg Leu Ile His Glu Asn Leu Pro Lys Phe Phe Asp Asn
195 200 205
Val Ile Ser Phe Asn Lys Leu Lys Glu Gly Phe Pro Glu Leu Lys Phe
210 215 220
Asp Lys Val Lys Glu Asp Leu Glu Val Asp Tyr Asp Leu Lys His Ala
225 230 235 240
Phe Glu Ile Glu Tyr Phe Val Asn Phe Val Thr Gln Ala Gly Ile Asp
245 250 255
Gln Tyr Asn Tyr Leu Leu Gly Gly Lys Thr Leu Glu Asp Gly Thr Lys
260 265 270
Lys Gln Gly Met Asn Glu Gln Ile Asn Leu Phe Lys Gln Gln Gln Thr
275 280 285
Arg Asp Lys Ala Arg Gln Ile Pro Lys Leu Ile Pro Leu Phe Lys Gln
290 295 300
Ile Leu Ser Glu Arg Thr Glu Ser Gln Ser Phe Ile Pro Lys Gln Phe
305 310 315 320
Glu Ser Asp Gln Glu Leu Phe Asp Ser Leu Gln Lys Leu His Asn Asn
325 330 335
Cys Gln Asp Lys Phe Thr Val Leu Gln Gln Ala Ile Leu Gly Leu Ala
340 345 350
Glu Ala Asp Leu Lys Lys Val Phe Ile Lys Thr Ser Asp Leu Asn Ala
355 360 365
Leu Ser Asn Thr Ile Phe Gly Asn Tyr Ser Val Phe Ser Asp Ala Leu
370 375 380
Asn Leu Tyr Lys Glu Ser Leu Lys Thr Lys Lys Ala Gln Glu Ala Phe
385 390 395 400
Glu Lys Leu Pro Ala His Ser Ile His Asp Leu Ile Gln Tyr Leu Glu
405 410 415
Gln Phe Asn Ser Ser Leu Asp Ala Glu Lys Gln Gln Ser Thr Asp Thr
420 425 430
Val Leu Asn Tyr Phe Ile Lys Thr Asp Glu Leu Tyr Ser Arg Phe Ile
435 440 445
Lys Ser Thr Ser Glu Ala Phe Thr Gln Val Gln Pro Leu Phe Glu Leu
450 455 460
Glu Ala Leu Ser Ser Lys Arg Arg Pro Pro Glu Ser Glu Asp Glu Gly
465 470 475 480
Ala Lys Gly Gln Glu Gly Phe Glu Gln Ile Lys Arg Ile Lys Ala Tyr
485 490 495
Leu Asp Thr Leu Met Glu Ala Val His Phe Ala Lys Pro Leu Tyr Leu
500 505 510
Val Lys Gly Arg Lys Met Ile Glu Gly Leu Asp Lys Asp Gln Ser Phe
515 520 525
Tyr Glu Ala Phe Glu Met Ala Tyr Gln Glu Leu Glu Ser Leu Ile Ile
530 535 540
Pro Ile Tyr Asn Lys Ala Arg Ser Tyr Leu Ser Arg Lys Pro Phe Lys
545 550 555 560
Ala Asp Lys Phe Lys Ile Asn Phe Asp Asn Asn Thr Leu Leu Ser Gly
565 570 575
Trp Asp Ala Asn Lys Glu Thr Ala Asn Ala Ser Ile Leu Phe Lys Lys
580 585 590
Asp Gly Leu Tyr Tyr Leu Gly Ile Met Pro Lys Gly Lys Thr Phe Leu
595 600 605
Phe Asp Tyr Phe Val Ser Ser Glu Asp Ser Glu Lys Leu Lys Gln Arg
610 615 620
Arg Gln Lys Thr Ala Glu Glu Ala Leu Ala Gln Asp Gly Glu Ser Tyr
625 630 635 640
Phe Glu Lys Ile Arg Tyr Lys Leu Leu Pro Gly Ala Ser Lys Met Leu
645 650 655
Pro Lys Val Phe Phe Ser Asn Lys Asn Ile Gly Phe Tyr Asn Pro Ser
660 665 670
Asp Asp Ile Leu Arg Ile Arg Asn Thr Ala Ser His Thr Lys Asn Gly
675 680 685
Thr Pro Gln Lys Gly His Ser Lys Val Glu Phe Asn Leu Asn Asp Cys
690 695 700
His Lys Met Ile Asp Phe Phe Lys Ser Ser Ile Gln Lys His Pro Glu
705 710 715 720
Trp Gly Ser Phe Gly Phe Thr Phe Ser Asp Thr Ser Asp Phe Glu Asp
725 730 735
Met Ser Ala Phe Tyr Arg Glu Val Glu Asn Gln Gly Tyr Val Ile Ser
740 745 750
Phe Asp Lys Ile Lys Glu Thr Tyr Ile Gln Ser Gln Val Glu Gln Gly
755 760 765
Asn Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Pro Tyr Ser
770 775 780
Lys Gly Lys Pro Asn Leu His Thr Leu Tyr Trp Lys Ala Leu Phe Glu
785 790 795 800
Glu Ala Asn Leu Asn Asn Val Val Ala Lys Leu Asn Gly Glu Ala Glu
805 810 815
Ile Phe Phe Arg Arg His Ser Ile Lys Ala Ser Asp Lys Val Val His
820 825 830
Pro Ala Asn Gln Ala Ile Asp Asn Lys Asn Pro His Thr Glu Lys Thr
835 840 845
Gln Ser Thr Phe Glu Tyr Asp Leu Val Lys Asp Lys Arg Tyr Thr Gln
850 855 860
Asp Lys Phe Phe Phe His Val Pro Ile Ser Leu Asn Phe Lys Ala Gln
865 870 875 880
Gly Val Ser Lys Phe Asn Asp Lys Val Asn Gly Phe Leu Lys Gly Asn
885 890 895
Pro Asp Val Asn Ile Ile Gly Ile Asp Arg Gly Glu Arg His Leu Leu
900 905 910
Tyr Phe Thr Val Val Asn Gln Lys Gly Glu Ile Leu Val Gln Glu Ser
915 920 925
Leu Asn Thr Leu Met Ser Asp Lys Gly His Val Asn Asp Tyr Gln Gln
930 935 940
Lys Leu Asp Lys Lys Glu Gln Glu Arg Asp Ala Ala Arg Lys Ser Trp
945 950 955 960
Thr Thr Val Glu Asn Ile Lys Glu Leu Lys Glu Gly Tyr Leu Ser His
965 970 975
Val Val His Lys Leu Ala His Leu Ile Ile Lys Tyr Asn Ala Ile Val
980 985 990
Cys Leu Glu Asp Leu Asn Phe Gly Phe Lys Arg Gly Arg Phe Lys Val
995 1000 1005
Glu Lys Gln Val Tyr Gln Lys Phe Glu Lys Ala Leu Ile Asp Lys
1010 1015 1020
Leu Asn Tyr Leu Val Phe Lys Glu Lys Glu Leu Gly Glu Val Gly
1025 1030 1035
His Tyr Leu Thr Ala Tyr Gln Leu Thr Ala Pro Phe Glu Ser Phe
1040 1045 1050
Lys Lys Leu Gly Lys Gln Ser Gly Ile Leu Phe Tyr Val Pro Ala
1055 1060 1065
Asp Tyr Thr Ser Lys Ile Asp Pro Thr Thr Gly Phe Val Asn Phe
1070 1075 1080
Leu Asp Leu Arg Tyr Gln Ser Val Glu Lys Ala Lys Gln Leu Leu
1085 1090 1095
Ser Asp Phe Asn Ala Ile Arg Phe Asn Ser Val Gln Asn Tyr Phe
1100 1105 1110
Glu Phe Glu Ile Asp Tyr Lys Lys Leu Thr Pro Lys Arg Lys Val
1115 1120 1125
Gly Thr Gln Ser Lys Trp Val Ile Cys Thr Tyr Gly Asp Val Arg
1130 1135 1140
Tyr Gln Asn Arg Arg Asn Gln Lys Gly His Trp Glu Thr Glu Glu
1145 1150 1155
Val Asn Val Thr Glu Lys Leu Lys Ala Leu Phe Ala Ser Asp Ser
1160 1165 1170
Lys Thr Thr Thr Val Ile Asp Tyr Ala Asn Asp Asp Asn Leu Ile
1175 1180 1185
Asp Val Ile Leu Glu Gln Asp Lys Ala Ser Phe Phe Lys Glu Leu
1190 1195 1200
Leu Trp Leu Leu Lys Leu Thr Met Thr Leu Arg His Ser Lys Ile
1205 1210 1215
Lys Ser Glu Asp Asp Phe Ile Leu Ser Pro Val Lys Asn Glu Gln
1220 1225 1230
Gly Glu Phe Tyr Asp Ser Arg Lys Ala Gly Glu Val Trp Pro Lys
1235 1240 1245
Asp Ala Asp Ala Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Leu
1250 1255 1260
Trp Asn Leu Gln Gln Ile Asn Gln Trp Glu Lys Gly Lys Thr Leu
1265 1270 1275
Asn Leu Ala Ile Lys Asn Gln Asp Trp Phe Ser Phe Ile Gln Glu
1280 1285 1290
Lys Pro Tyr Gln Glu
1295
<210> 37
<211> 1242
<212> PRT
<213> 拟杆菌目细菌
<220>
<221> MISC_FEATURE
<222> (1)..(1242)
<223> Genbank KXB38146 Cpf1
<400> 37
Met Lys Lys Phe Thr Asn Leu Tyr Pro Val Gln Lys Thr Leu Arg Phe
1 5 10 15
Glu Leu Ile Pro Gln Gly Asn Thr Ser Lys His Leu Cys Lys Ile Ile
20 25 30
Gln Glu Asp Glu Gln Ile Ala Glu Asp Ser Gln Glu Val Lys Lys Leu
35 40 45
Leu Asp Arg Tyr His Lys Glu Phe Ile Ala Ile Ala Leu Ser Ser Phe
50 55 60
Pro Thr Ser Pro Leu Ala Lys Glu Ile Ile Pro Lys Leu Lys Glu Phe
65 70 75 80
Ala Gln Ile Arg Ala Thr Gly Asp Ala Lys Gln Ile Ser Thr Ile Gln
85 90 95
Asp Glu Leu Arg Glu Leu Val Val Lys Gly Phe Lys Gly Glu Gly Glu
100 105 110
Gln Glu Arg Arg Tyr Lys Ile Leu Ile Gly Ala Lys Gly Asn Pro Asn
115 120 125
Ala Asp Glu Leu Phe Asn Thr Glu Leu Ile Asn Phe Leu Lys Asp Pro
130 135 140
Ala Glu Gln Ala Leu Val Lys Lys Phe Gln Lys His Thr Gly Tyr Phe
145 150 155 160
Leu Gly Phe Asn Glu Asn Arg Lys Asn Met Tyr Ser Ala Lys Ala Gln
165 170 175
Ser Thr Ala Ile Ala Tyr Arg Leu Ile His Glu Asn Leu Pro Arg Phe
180 185 190
Leu Asp Asn Ile Thr Thr Tyr Glu Lys Val Lys Thr Tyr Leu Lys Glu
195 200 205
Glu Ile Pro Gln Leu Glu Lys Glu Leu Val Arg Ala Gly Ala Ser Leu
210 215 220
Val Ser His Val Asp Ser Val Phe Thr Ile Asp Phe Phe Leu Glu Val
225 230 235 240
Phe Thr Gln Ser Gly Ile Asp Gln Tyr Asn Ala Leu Ile Gly Lys Ile
245 250 255
Val Asn Gln Glu Gln Gly Glu Val Lys Gly Leu Asn Glu Arg Ile Asn
260 265 270
Leu Tyr Asn Gln Gln His Lys Gln Glu Ala Lys Leu Pro Leu Phe Lys
275 280 285
Pro Leu Tyr Lys Gln Ile Leu Ser Asp Arg Glu Gln Leu Ser Trp Leu
290 295 300
Ala Glu Ala Tyr Asn Ser Asp Lys Asp Leu Leu Asp Ser Ile Gln Lys
305 310 315 320
Tyr Tyr Gln Leu Leu Ile Asp Asn Gln Ile Phe Glu Arg Ile Pro Arg
325 330 335
Leu Met His Thr Leu Glu Lys Ala Pro Leu Asp Lys Ile Trp Ile Thr
340 345 350
Tyr Asp Thr Gln Leu Thr Ser Ile Ser Asn Thr Leu Tyr Gly Ser Trp
355 360 365
Arg Val Ile Gly Glu Ala Leu Gly Arg Asn Ala Lys Ser Glu Lys Glu
370 375 380
Arg Lys Ser Ser Gln Lys Lys Ala Leu Asn Tyr Ser Leu Glu Ser Ile
385 390 395 400
Asn Gln Ala Ile Ala Lys Met Pro Ser Asp Glu Glu Leu Pro Pro Ile
405 410 415
Gln Lys Tyr Phe Ile Ala Leu Gly Ser Asn Pro Ser Lys Lys Asp Ala
420 425 430
Ser Ser Gly Thr Leu Val Asp Lys Val Arg Ser Ser Tyr Lys Ala Cys
435 440 445
Gln Asp Ile Leu Thr Asn Pro Asp His Thr Gly Lys Lys Leu Ile Gln
450 455 460
Asp Lys Lys Gln Val Asp Leu Leu Lys Gln Leu Leu Asp Asp Leu Leu
465 470 475 480
Ile Leu Gln Arg Phe Ile Lys Pro Leu Leu Tyr Ser Asn Asn Glu Asn
485 490 495
Glu Thr His Lys Asp Glu Val Phe Tyr Thr Glu Leu Thr Asp Ile Met
500 505 510
Asp Leu Leu Asn Pro Ile Val Gly Leu Tyr Asn Lys Val Arg Asn Tyr
515 520 525
Leu Thr Gln Lys Pro Tyr Ser Thr Glu Lys Phe Lys Ile Asn Phe Lys
530 535 540
Ser Ser Ser Leu Leu Ala Gly Trp Asp Arg Asn Lys Glu Lys Asp Asn
545 550 555 560
Leu Gly Val Ile Leu Lys Arg Glu Asp Lys Tyr Tyr Leu Ala Ile Met
565 570 575
Asp Lys Ala His Asn Ala Thr Phe Lys Asn Lys Ser Leu Pro Thr Gln
580 585 590
Gly Glu Cys Tyr Glu Lys Met Glu Tyr Lys Leu Leu Pro Gly Ala Asn
595 600 605
Lys Met Leu Pro Lys Val Tyr Ile Thr Ser Lys Lys Gly Ile Glu Ser
610 615 620
Phe His Pro Ser Glu Glu Leu Gln Lys Lys Tyr Lys Leu Gly Thr His
625 630 635 640
Lys Lys Gly Ala Ser Phe Asn Leu Ser Asp Met Arg Ala Leu Ile Asp
645 650 655
Tyr Phe Lys Glu Ser Leu Glu Lys His Glu Glu His Ser Gln Phe Gly
660 665 670
Phe His Phe Ser Asp Thr Ser Thr Tyr Glu Asp Ile Ser Gly Phe Tyr
675 680 685
Arg Glu Val Glu Gln Gln Ala Tyr Lys Ile Thr Phe Arg Lys Val Ser
690 695 700
Val Glu Tyr Ile Asp Gln Leu Val Asn Glu Gly Lys Leu Tyr Leu Phe
705 710 715 720
Gln Ile Tyr Asn Lys Asp Phe Ser Pro Tyr Ser Lys Gly Thr Pro Asn
725 730 735
Leu His Thr Leu Tyr Trp Lys Met Leu Phe Asp Pro Ala Asn Leu Gln
740 745 750
Asp Ile Val Tyr Lys Leu Asn Gly Glu Ala Glu Val Phe Phe Arg Lys
755 760 765
Lys Ser Leu Gln Tyr Asp Arg Pro Thr His Pro Lys Gly Gln Pro Ile
770 775 780
Asn Lys Lys Ser Leu Leu Asn Glu Gly Glu Thr Ser Leu Phe Asp Tyr
785 790 795 800
Asp Leu Ile Lys Asp Arg Arg Phe Thr Val Asp Lys Phe Gln Phe His
805 810 815
Val Pro Ile Thr Met Asn Phe Lys Ala Thr Gln Gly Thr Lys Val Asn
820 825 830
Gln Met Val Gln Glu Glu Val Lys Lys Ser Lys Gly Phe His Leu Ile
835 840 845
Gly Ile Asp Arg Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Ile Asn
850 855 860
Glu Arg Gly Glu Ile Ile Glu Gln Cys Ser Leu Asn Lys Ile Val Asn
865 870 875 880
Thr Tyr Gln Glu Lys Glu His Thr Val Asp Tyr Lys Ala Leu Leu Glu
885 890 895
Lys Arg Ser Gln Ser Arg Leu Glu Glu Arg Lys Ser Trp Gln Thr Ile
900 905 910
Glu Asn Ile Lys Glu Leu Lys Gly Gly Tyr Leu Ser Gln Val Val His
915 920 925
Lys Ile Ala Gln Leu Met Ile Lys Tyr Asn Ala Ile Ala Val Leu Glu
930 935 940
Asp Leu Asn Phe Gly Phe Ile Arg Thr Arg Lys Lys Phe Glu Phe Ser
945 950 955 960
Val Tyr Gln Glu Phe Glu Lys Lys Leu Ile Asp Lys Leu Gly Tyr Val
965 970 975
Val Asp Lys Lys Ala Pro Ile Gln Gln Glu Gly Gly Leu Leu Gln Ala
980 985 990
Tyr Gln Leu Thr Ala Pro Phe Lys Ser Phe Arg Glu Met Gly Lys Gln
995 1000 1005
Asn Gly Phe Leu Phe Tyr Val Pro Ala Trp Asn Thr Ser Ala Ile
1010 1015 1020
Asp Pro Arg Thr Gly Phe Val Asn Leu Leu Asp Thr Arg Tyr Glu
1025 1030 1035
Ser Ile Ala Lys Thr Lys Glu Leu Ile Lys Lys Leu Lys Asp Ile
1040 1045 1050
Arg Tyr Asn Ser Gln Lys Asp Trp Phe Glu Ile Asp Leu Asp Tyr
1055 1060 1065
Asn Ala Phe Gly Asn Arg Ala Lys Gly Ser Arg Ser Lys Trp Arg
1070 1075 1080
Leu Cys Ser Tyr Gly Glu Arg Ile Glu His Thr Arg Lys Gln Asp
1085 1090 1095
Ser Asn Gly Gln Glu Glu Ser Asp Ser Met Val Val Leu Thr Glu
1100 1105 1110
Ala Phe Lys Asp Val Phe Thr Lys Tyr Gln Ile Asp Tyr Arg Glu
1115 1120 1125
Asn Leu Lys Glu Gln Leu Leu Leu Gln Ser Asp Lys Ala Phe Phe
1130 1135 1140
Val Asp Phe Leu Ser Leu Leu Arg Leu Thr Leu Gln Leu Arg Asn
1145 1150 1155
Ser Leu Ser Asn Ser Leu Ile Asp Tyr Ile Leu Ser Pro Val Ala
1160 1165 1170
Asp Glu Asn Gly Glu Phe Phe Asp Ser Arg Lys Ala Leu Ser Asn
1175 1180 1185
Glu Pro Gln Asp Ala Asp Ala Asn Gly Ala Tyr His Ile Ala Leu
1190 1195 1200
Lys Gly Leu Trp Val Leu Asp Lys Ile Arg Lys Thr Glu Lys Val
1205 1210 1215
Thr Pro Ala Lys Leu Ala Leu Ser Asn Gln Glu Trp Leu Ser Phe
1220 1225 1230
Ala Gln Glu Lys Pro Phe Phe Asn Glu
1235 1240
<210> 38
<211> 1300
<212> PRT
<213> 土拉弗朗西斯菌
<220>
<221> MISC_FEATURE
<222> (1)..(1300)
<223> Genbank WP_003040289 Cpf1
<400> 38
Met Ser Ile Tyr Gln Glu Phe Val Asn Lys Tyr Ser Leu Ser Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Glu Asn Ile Lys
20 25 30
Ala Arg Gly Leu Ile Leu Asp Asp Glu Lys Arg Ala Lys Asp Tyr Lys
35 40 45
Lys Ala Lys Gln Ile Ile Asp Lys Tyr His Gln Phe Phe Ile Glu Glu
50 55 60
Ile Leu Ser Ser Val Cys Ile Ser Glu Asp Leu Leu Gln Asn Tyr Ser
65 70 75 80
Asp Val Tyr Phe Lys Leu Lys Lys Ser Asp Asp Asp Asn Leu Gln Lys
85 90 95
Asp Phe Lys Ser Ala Lys Asp Thr Ile Lys Lys Gln Ile Ser Glu Tyr
100 105 110
Ile Lys Asp Ser Glu Lys Phe Lys Asn Leu Phe Asn Gln Asn Leu Ile
115 120 125
Asp Ala Lys Lys Gly Gln Glu Ser Asp Leu Ile Leu Trp Leu Lys Gln
130 135 140
Ser Lys Asp Asn Gly Ile Glu Leu Phe Lys Ala Asn Ser Asp Ile Thr
145 150 155 160
Asp Ile Asp Glu Ala Leu Glu Ile Ile Lys Ser Phe Lys Gly Trp Thr
165 170 175
Thr Tyr Phe Lys Gly Phe His Glu Asn Arg Lys Asn Val Tyr Ser Ser
180 185 190
Asn Asp Ile Pro Thr Ser Ile Ile Tyr Arg Ile Val Asp Asp Asn Leu
195 200 205
Pro Lys Phe Leu Glu Asn Lys Ala Lys Tyr Glu Ser Leu Lys Asp Lys
210 215 220
Ala Pro Glu Ala Ile Asn Tyr Glu Gln Ile Lys Lys Asp Leu Ala Glu
225 230 235 240
Glu Leu Thr Phe Asp Ile Asp Tyr Lys Thr Ser Glu Val Asn Gln Arg
245 250 255
Val Phe Ser Leu Asp Glu Val Phe Glu Ile Ala Asn Phe Asn Asn Tyr
260 265 270
Leu Asn Gln Ser Gly Ile Thr Lys Phe Asn Thr Ile Ile Gly Gly Lys
275 280 285
Phe Val Asn Gly Glu Asn Thr Lys Arg Lys Gly Ile Asn Glu Tyr Ile
290 295 300
Asn Leu Tyr Ser Gln Gln Ile Asn Asp Lys Thr Leu Lys Lys Tyr Lys
305 310 315 320
Met Ser Val Leu Phe Lys Gln Ile Leu Ser Asp Thr Glu Ser Lys Ser
325 330 335
Phe Val Ile Asp Lys Leu Glu Asp Asp Ser Asp Val Val Thr Thr Met
340 345 350
Gln Ser Phe Tyr Glu Gln Ile Ala Ala Phe Lys Thr Val Glu Glu Lys
355 360 365
Ser Ile Lys Glu Thr Leu Ser Leu Leu Phe Asp Asp Leu Lys Ala Gln
370 375 380
Lys Leu Asp Leu Ser Lys Ile Tyr Phe Lys Asn Asp Lys Ser Leu Thr
385 390 395 400
Asp Leu Ser Gln Gln Val Phe Asp Asp Tyr Ser Val Ile Gly Thr Ala
405 410 415
Val Leu Glu Tyr Ile Thr Gln Gln Ile Ala Pro Lys Asn Leu Asp Asn
420 425 430
Pro Ser Lys Lys Glu Gln Glu Leu Ile Ala Lys Lys Thr Glu Lys Ala
435 440 445
Lys Tyr Leu Ser Leu Glu Thr Ile Lys Leu Ala Leu Glu Glu Phe Asn
450 455 460
Lys His Arg Asp Ile Asp Lys Gln Cys Arg Phe Glu Glu Ile Leu Ala
465 470 475 480
Asn Phe Ala Ala Ile Pro Met Ile Phe Asp Glu Ile Ala Gln Asn Lys
485 490 495
Asp Asn Leu Ala Gln Ile Ser Ile Lys Tyr Gln Asn Gln Gly Lys Lys
500 505 510
Asp Leu Leu Gln Ala Ser Ala Glu Asp Asp Val Lys Ala Ile Lys Asp
515 520 525
Leu Leu Asp Gln Thr Asn Asn Leu Leu His Lys Leu Lys Ile Phe His
530 535 540
Ile Ser Gln Ser Glu Asp Lys Ala Asn Ile Leu Asp Lys Asp Glu His
545 550 555 560
Phe Tyr Leu Val Phe Glu Glu Cys Tyr Phe Glu Leu Ala Asn Ile Val
565 570 575
Pro Leu Tyr Asn Lys Ile Arg Asn Tyr Ile Thr Gln Lys Pro Tyr Ser
580 585 590
Asp Glu Lys Phe Lys Leu Asn Phe Glu Asn Ser Thr Leu Ala Asn Gly
595 600 605
Trp Asp Lys Asn Lys Glu Pro Asp Asn Thr Ala Ile Leu Phe Ile Lys
610 615 620
Asp Asp Lys Tyr Tyr Leu Gly Val Met Asn Lys Lys Asn Asn Lys Ile
625 630 635 640
Phe Asp Asp Lys Ala Ile Lys Glu Asn Lys Gly Glu Gly Tyr Lys Lys
645 650 655
Ile Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro Lys Val
660 665 670
Phe Phe Ser Ala Lys Ser Ile Lys Phe Tyr Asn Pro Ser Glu Asp Ile
675 680 685
Leu Arg Ile Arg Asn His Ser Thr His Thr Lys Asn Gly Ser Pro Gln
690 695 700
Lys Gly Tyr Glu Lys Phe Glu Phe Asn Ile Glu Asp Cys Arg Lys Phe
705 710 715 720
Ile Asp Phe Tyr Lys Gln Ser Ile Ser Lys His Pro Glu Trp Lys Asp
725 730 735
Phe Gly Phe Arg Phe Ser Asp Thr Gln Arg Tyr Asn Ser Ile Asp Glu
740 745 750
Phe Tyr Arg Glu Val Glu Asn Gln Gly Tyr Lys Leu Thr Phe Glu Asn
755 760 765
Ile Ser Glu Ser Tyr Ile Asp Ser Val Val Asn Gln Gly Lys Leu Tyr
770 775 780
Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Ala Tyr Ser Lys Gly Arg
785 790 795 800
Pro Asn Leu His Thr Leu Tyr Trp Lys Ala Leu Phe Asp Glu Arg Asn
805 810 815
Leu Gln Asp Val Val Tyr Lys Leu Asn Gly Glu Ala Glu Leu Phe Tyr
820 825 830
Arg Lys Gln Ser Ile Pro Lys Lys Ile Thr His Pro Ala Lys Glu Ala
835 840 845
Ile Ala Asn Lys Asn Lys Asp Asn Pro Lys Lys Glu Ser Val Phe Glu
850 855 860
Tyr Asp Leu Ile Lys Asp Lys Arg Phe Thr Glu Asp Lys Phe Phe Phe
865 870 875 880
His Cys Pro Ile Thr Ile Asn Phe Lys Ser Ser Gly Ala Asn Lys Phe
885 890 895
Asn Asp Glu Ile Asn Leu Leu Leu Lys Glu Lys Ala Asn Asp Val His
900 905 910
Ile Leu Ser Ile Asp Arg Gly Glu Arg His Leu Ala Tyr Tyr Thr Leu
915 920 925
Val Asp Gly Lys Gly Asn Ile Ile Lys Gln Asp Thr Phe Asn Ile Ile
930 935 940
Gly Asn Asp Arg Met Lys Thr Asn Tyr His Asp Lys Leu Ala Ala Ile
945 950 955 960
Glu Lys Asp Arg Asp Ser Ala Arg Lys Asp Trp Lys Lys Ile Asn Asn
965 970 975
Ile Lys Glu Met Lys Glu Gly Tyr Leu Ser Gln Val Val His Glu Ile
980 985 990
Ala Lys Leu Val Ile Glu Tyr Asn Ala Ile Val Val Phe Glu Asp Leu
995 1000 1005
Asn Phe Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Val
1010 1015 1020
Tyr Gln Lys Leu Glu Lys Met Leu Ile Glu Lys Leu Asn Tyr Leu
1025 1030 1035
Val Phe Lys Asp Asn Glu Phe Asp Lys Thr Gly Gly Val Leu Arg
1040 1045 1050
Ala Tyr Gln Leu Thr Ala Pro Phe Glu Thr Phe Lys Lys Met Gly
1055 1060 1065
Lys Gln Thr Gly Ile Ile Tyr Tyr Val Pro Ala Gly Phe Thr Ser
1070 1075 1080
Lys Ile Cys Pro Val Thr Gly Phe Val Asn Gln Leu Tyr Pro Lys
1085 1090 1095
Tyr Glu Ser Val Ser Lys Ser Gln Glu Phe Phe Ser Lys Phe Asp
1100 1105 1110
Lys Ile Cys Tyr Asn Leu Asp Lys Gly Tyr Phe Glu Phe Ser Phe
1115 1120 1125
Asp Tyr Lys Asn Phe Gly Asp Lys Ala Ala Lys Gly Lys Trp Thr
1130 1135 1140
Ile Ala Ser Phe Gly Ser Arg Leu Ile Asn Phe Arg Asn Ser Asp
1145 1150 1155
Lys Asn His Asn Trp Asp Thr Arg Glu Val Tyr Pro Thr Lys Glu
1160 1165 1170
Leu Glu Lys Leu Leu Lys Asp Tyr Ser Ile Glu Tyr Gly His Gly
1175 1180 1185
Glu Cys Ile Lys Ala Ala Ile Cys Gly Glu Ser Asp Lys Lys Phe
1190 1195 1200
Phe Ala Lys Leu Thr Ser Val Leu Asn Thr Ile Leu Gln Met Arg
1205 1210 1215
Asn Ser Lys Thr Gly Thr Glu Leu Asp Tyr Leu Ile Ser Pro Val
1220 1225 1230
Ala Asp Val Asn Gly Asn Phe Phe Asp Ser Arg Gln Ala Pro Lys
1235 1240 1245
Asn Met Pro Gln Asp Ala Asp Ala Asn Gly Ala Tyr His Ile Gly
1250 1255 1260
Leu Lys Gly Leu Met Leu Leu Gly Arg Ile Lys Asn Asn Gln Glu
1265 1270 1275
Gly Lys Lys Leu Asn Leu Val Ile Lys Asn Glu Glu Tyr Phe Glu
1280 1285 1290
Phe Val Gln Asn Arg Asn Asn
1295 1300
<210> 39
<211> 1310
<212> PRT
<213> 孔氏创伤球菌
<220>
<221> MISC_FEATURE
<222> (1)..(1310)
<223> Genbank WP_005398606 Cpf1
<400> 39
Met Phe Glu Lys Leu Ser Asn Ile Val Ser Ile Ser Lys Thr Ile Arg
1 5 10 15
Phe Lys Leu Ile Pro Val Gly Lys Thr Leu Glu Asn Ile Glu Lys Leu
20 25 30
Gly Lys Leu Glu Lys Asp Phe Glu Arg Ser Asp Phe Tyr Pro Ile Leu
35 40 45
Lys Asn Ile Ser Asp Asp Tyr Tyr Arg Gln Tyr Ile Lys Glu Lys Leu
50 55 60
Ser Asp Leu Asn Leu Asp Trp Gln Lys Leu Tyr Asp Ala His Glu Leu
65 70 75 80
Leu Asp Ser Ser Lys Lys Glu Ser Gln Lys Asn Leu Glu Met Ile Gln
85 90 95
Ala Gln Tyr Arg Lys Val Leu Phe Asn Ile Leu Ser Gly Glu Leu Asp
100 105 110
Lys Ser Gly Glu Lys Asn Ser Lys Asp Leu Ile Lys Asn Asn Lys Ala
115 120 125
Leu Tyr Gly Lys Leu Phe Lys Lys Gln Phe Ile Leu Glu Val Leu Pro
130 135 140
Asp Phe Val Asn Asn Asn Asp Ser Tyr Ser Glu Glu Asp Leu Glu Gly
145 150 155 160
Leu Asn Leu Tyr Ser Lys Phe Thr Thr Arg Leu Lys Asn Phe Trp Glu
165 170 175
Thr Arg Lys Asn Val Phe Thr Asp Lys Asp Ile Val Thr Ala Ile Pro
180 185 190
Phe Arg Ala Val Asn Glu Asn Phe Gly Phe Tyr Tyr Asp Asn Ile Lys
195 200 205
Ile Phe Asn Lys Asn Ile Glu Tyr Leu Glu Asn Lys Ile Pro Asn Leu
210 215 220
Glu Asn Glu Leu Lys Glu Ala Asp Ile Leu Asp Asp Asn Arg Ser Val
225 230 235 240
Lys Asp Tyr Phe Thr Pro Asn Gly Phe Asn Tyr Val Ile Thr Gln Asp
245 250 255
Gly Ile Asp Val Tyr Gln Ala Ile Arg Gly Gly Phe Thr Lys Glu Asn
260 265 270
Gly Glu Lys Val Gln Gly Ile Asn Glu Ile Leu Asn Leu Thr Gln Gln
275 280 285
Gln Leu Arg Arg Lys Pro Glu Thr Lys Asn Val Lys Leu Gly Val Leu
290 295 300
Thr Lys Leu Arg Lys Gln Ile Leu Glu Tyr Ser Glu Ser Thr Ser Phe
305 310 315 320
Leu Ile Asp Gln Ile Glu Asp Asp Asn Asp Leu Val Asp Arg Ile Asn
325 330 335
Lys Phe Asn Val Ser Phe Phe Glu Ser Thr Glu Val Ser Pro Ser Leu
340 345 350
Phe Glu Gln Ile Glu Arg Leu Tyr Asn Ala Leu Lys Ser Ile Lys Lys
355 360 365
Glu Glu Val Tyr Ile Asp Ala Arg Asn Thr Gln Lys Phe Ser Gln Met
370 375 380
Leu Phe Gly Gln Trp Asp Val Ile Arg Arg Gly Tyr Thr Val Lys Ile
385 390 395 400
Thr Glu Gly Ser Lys Glu Glu Lys Lys Lys Tyr Lys Glu Tyr Leu Glu
405 410 415
Leu Asp Glu Thr Ser Lys Ala Lys Arg Tyr Leu Asn Ile Arg Glu Ile
420 425 430
Glu Glu Leu Val Asn Leu Val Glu Gly Phe Glu Glu Val Asp Val Phe
435 440 445
Ser Val Leu Leu Glu Lys Phe Lys Met Asn Asn Ile Glu Arg Ser Glu
450 455 460
Phe Glu Ala Pro Ile Tyr Gly Ser Pro Ile Lys Leu Glu Ala Ile Lys
465 470 475 480
Glu Tyr Leu Glu Lys His Leu Glu Glu Tyr His Lys Trp Lys Leu Leu
485 490 495
Leu Ile Gly Asn Asp Asp Leu Asp Thr Asp Glu Thr Phe Tyr Pro Leu
500 505 510
Leu Asn Glu Val Ile Ser Asp Tyr Tyr Ile Ile Pro Leu Tyr Asn Leu
515 520 525
Thr Arg Asn Tyr Leu Thr Arg Lys His Ser Asp Lys Asp Lys Ile Lys
530 535 540
Val Asn Phe Asp Phe Pro Thr Leu Ala Asp Gly Trp Ser Glu Ser Lys
545 550 555 560
Ile Ser Asp Asn Arg Ser Ile Ile Leu Arg Lys Gly Gly Tyr Tyr Tyr
565 570 575
Leu Gly Ile Leu Ile Asp Asn Lys Leu Leu Ile Asn Lys Lys Asn Lys
580 585 590
Ser Lys Lys Ile Tyr Glu Ile Leu Ile Tyr Asn Gln Ile Pro Glu Phe
595 600 605
Ser Lys Ser Ile Pro Asn Tyr Pro Phe Thr Lys Lys Val Lys Glu His
610 615 620
Phe Lys Asn Asn Val Ser Asp Phe Gln Leu Ile Asp Gly Tyr Val Ser
625 630 635 640
Pro Leu Ile Ile Thr Lys Glu Ile Tyr Asp Ile Lys Lys Glu Lys Lys
645 650 655
Tyr Lys Lys Asp Phe Tyr Lys Asp Asn Asn Thr Asn Lys Asn Tyr Leu
660 665 670
Tyr Thr Ile Tyr Lys Trp Ile Glu Phe Cys Lys Gln Phe Leu Tyr Lys
675 680 685
Tyr Lys Gly Pro Asn Lys Glu Ser Tyr Lys Glu Met Tyr Asp Phe Ser
690 695 700
Thr Leu Lys Asp Thr Ser Leu Tyr Val Asn Leu Asn Asp Phe Tyr Ala
705 710 715 720
Asp Val Asn Ser Cys Ala Tyr Arg Val Leu Phe Asn Lys Ile Asp Glu
725 730 735
Asn Thr Ile Asp Asn Ala Val Glu Asp Gly Lys Leu Leu Leu Phe Gln
740 745 750
Ile Tyr Asn Lys Asp Phe Ser Pro Glu Ser Lys Gly Lys Lys Asn Leu
755 760 765
His Thr Leu Tyr Trp Leu Ser Met Phe Ser Glu Glu Asn Leu Arg Thr
770 775 780
Arg Lys Leu Lys Leu Asn Gly Gln Ala Glu Ile Phe Tyr Arg Lys Lys
785 790 795 800
Leu Glu Lys Lys Pro Ile Ile His Lys Glu Gly Ser Ile Leu Leu Asn
805 810 815
Lys Ile Asp Lys Glu Gly Asn Thr Ile Pro Glu Asn Ile Tyr His Glu
820 825 830
Cys Tyr Arg Tyr Leu Asn Lys Lys Ile Gly Arg Glu Asp Leu Ser Asp
835 840 845
Glu Ala Ile Ala Leu Phe Asn Lys Asp Val Leu Lys Tyr Lys Glu Ala
850 855 860
Arg Phe Asp Ile Ile Lys Asp Arg Arg Tyr Ser Glu Ser Gln Phe Phe
865 870 875 880
Phe His Val Pro Ile Thr Phe Asn Trp Asp Ile Lys Thr Asn Lys Asn
885 890 895
Val Asn Gln Ile Val Gln Gly Met Ile Lys Asp Gly Glu Ile Lys His
900 905 910
Ile Ile Gly Ile Asp Arg Gly Glu Arg His Leu Leu Tyr Tyr Ser Val
915 920 925
Ile Asp Leu Glu Gly Asn Ile Val Glu Gln Gly Ser Leu Asn Thr Leu
930 935 940
Glu Gln Asn Arg Phe Asp Asn Ser Thr Val Lys Val Asp Tyr Gln Asn
945 950 955 960
Lys Leu Arg Thr Arg Glu Glu Asp Arg Asp Arg Ala Arg Lys Asn Trp
965 970 975
Thr Asn Ile Asn Lys Ile Lys Glu Leu Lys Asp Gly Tyr Leu Ser His
980 985 990
Val Val His Lys Leu Ser Arg Leu Ile Ile Lys Tyr Glu Ala Ile Val
995 1000 1005
Ile Met Glu Asn Leu Asn Gln Gly Phe Lys Arg Gly Arg Phe Lys
1010 1015 1020
Val Glu Arg Gln Val Tyr Gln Lys Phe Glu Leu Ala Leu Met Asn
1025 1030 1035
Lys Leu Ser Ala Leu Ser Phe Lys Glu Lys Tyr Asp Glu Arg Lys
1040 1045 1050
Asn Leu Glu Pro Ser Gly Ile Leu Asn Pro Ile Gln Ala Cys Tyr
1055 1060 1065
Pro Val Asp Ala Tyr Gln Glu Leu Gln Gly Gln Asn Gly Ile Val
1070 1075 1080
Phe Tyr Leu Pro Ala Ala Tyr Thr Ser Val Ile Asp Pro Val Thr
1085 1090 1095
Gly Phe Thr Asn Leu Phe Arg Leu Lys Ser Ile Asn Ser Ser Lys
1100 1105 1110
Tyr Glu Glu Phe Ile Lys Lys Phe Lys Asn Ile Tyr Phe Asp Asn
1115 1120 1125
Glu Glu Glu Asp Phe Lys Phe Ile Phe Asn Tyr Lys Asp Phe Ala
1130 1135 1140
Lys Ala Asn Leu Val Ile Leu Asn Asn Ile Lys Ser Lys Asp Trp
1145 1150 1155
Lys Ile Ser Thr Arg Gly Glu Arg Ile Ser Tyr Asn Ser Lys Lys
1160 1165 1170
Lys Glu Tyr Phe Tyr Val Gln Pro Thr Glu Phe Leu Ile Asn Lys
1175 1180 1185
Leu Lys Glu Leu Asn Ile Asp Tyr Glu Asn Ile Asp Ile Ile Pro
1190 1195 1200
Leu Ile Asp Asn Leu Glu Glu Lys Ala Lys Arg Lys Ile Leu Lys
1205 1210 1215
Ala Leu Phe Asp Thr Phe Lys Tyr Ser Val Gln Leu Arg Asn Tyr
1220 1225 1230
Asp Phe Glu Asn Asp Tyr Ile Ile Ser Pro Thr Ala Asp Asp Asn
1235 1240 1245
Gly Asn Tyr Tyr Asn Ser Asn Glu Ile Asp Ile Asp Lys Thr Asn
1250 1255 1260
Leu Pro Asn Asn Gly Asp Ala Asn Gly Ala Phe Asn Ile Ala Arg
1265 1270 1275
Lys Gly Leu Leu Leu Lys Asp Arg Ile Val Asn Ser Asn Glu Ser
1280 1285 1290
Lys Val Asp Leu Lys Ile Lys Asn Glu Asp Trp Ile Asn Phe Ile
1295 1300 1305
Ile Ser
1310
<210> 40
<211> 1257
<212> PRT
<213> 布氏普雷沃氏菌
<220>
<221> MISC_FEATURE
<222> (1)..(1257)
<223> Genbank WP_006283774 Cpf1
<400> 40
Met Gln Ile Asn Asn Leu Lys Ile Ile Tyr Met Lys Phe Thr Asp Phe
1 5 10 15
Thr Gly Leu Tyr Ser Leu Ser Lys Thr Leu Arg Phe Glu Leu Lys Pro
20 25 30
Ile Gly Lys Thr Leu Glu Asn Ile Lys Lys Ala Gly Leu Leu Glu Gln
35 40 45
Asp Gln His Arg Ala Asp Ser Tyr Lys Lys Val Lys Lys Ile Ile Asp
50 55 60
Glu Tyr His Lys Ala Phe Ile Glu Lys Ser Leu Ser Asn Phe Glu Leu
65 70 75 80
Lys Tyr Gln Ser Glu Asp Lys Leu Asp Ser Leu Glu Glu Tyr Leu Met
85 90 95
Tyr Tyr Ser Met Lys Arg Ile Glu Lys Thr Glu Lys Asp Lys Phe Ala
100 105 110
Lys Ile Gln Asp Asn Leu Arg Lys Gln Ile Ala Asp His Leu Lys Gly
115 120 125
Asp Glu Ser Tyr Lys Thr Ile Phe Ser Lys Asp Leu Ile Arg Lys Asn
130 135 140
Leu Pro Asp Phe Val Lys Ser Asp Glu Glu Arg Thr Leu Ile Lys Glu
145 150 155 160
Phe Lys Asp Phe Thr Thr Tyr Phe Lys Gly Phe Tyr Glu Asn Arg Glu
165 170 175
Asn Met Tyr Ser Ala Glu Asp Lys Ser Thr Ala Ile Ser His Arg Ile
180 185 190
Ile His Glu Asn Leu Pro Lys Phe Val Asp Asn Ile Asn Ala Phe Ser
195 200 205
Lys Ile Ile Leu Ile Pro Glu Leu Arg Glu Lys Leu Asn Gln Ile Tyr
210 215 220
Gln Asp Phe Glu Glu Tyr Leu Asn Val Glu Ser Ile Asp Glu Ile Phe
225 230 235 240
His Leu Asp Tyr Phe Ser Met Val Met Thr Gln Lys Gln Ile Glu Val
245 250 255
Tyr Asn Ala Ile Ile Gly Gly Lys Ser Thr Asn Asp Lys Lys Ile Gln
260 265 270
Gly Leu Asn Glu Tyr Ile Asn Leu Tyr Asn Gln Lys His Lys Asp Cys
275 280 285
Lys Leu Pro Lys Leu Lys Leu Leu Phe Lys Gln Ile Leu Ser Asp Arg
290 295 300
Ile Ala Ile Ser Trp Leu Pro Asp Asn Phe Lys Asp Asp Gln Glu Ala
305 310 315 320
Leu Asp Ser Ile Asp Thr Cys Tyr Lys Asn Leu Leu Asn Asp Gly Asn
325 330 335
Val Leu Gly Glu Gly Asn Leu Lys Leu Leu Leu Glu Asn Ile Asp Thr
340 345 350
Tyr Asn Leu Lys Gly Ile Phe Ile Arg Asn Asp Leu Gln Leu Thr Asp
355 360 365
Ile Ser Gln Lys Met Tyr Ala Ser Trp Asn Val Ile Gln Asp Ala Val
370 375 380
Ile Leu Asp Leu Lys Lys Gln Val Ser Arg Lys Lys Lys Glu Ser Ala
385 390 395 400
Glu Asp Tyr Asn Asp Arg Leu Lys Lys Leu Tyr Thr Ser Gln Glu Ser
405 410 415
Phe Ser Ile Gln Tyr Leu Asn Asp Cys Leu Arg Ala Tyr Gly Lys Thr
420 425 430
Glu Asn Ile Gln Asp Tyr Phe Ala Lys Leu Gly Ala Val Asn Asn Glu
435 440 445
His Glu Gln Thr Ile Asn Leu Phe Ala Gln Val Arg Asn Ala Tyr Thr
450 455 460
Ser Val Gln Ala Ile Leu Thr Thr Pro Tyr Pro Glu Asn Ala Asn Leu
465 470 475 480
Ala Gln Asp Lys Glu Thr Val Ala Leu Ile Lys Asn Leu Leu Asp Ser
485 490 495
Leu Lys Arg Leu Gln Arg Phe Ile Lys Pro Leu Leu Gly Lys Gly Asp
500 505 510
Glu Ser Asp Lys Asp Glu Arg Phe Tyr Gly Asp Phe Thr Pro Leu Trp
515 520 525
Glu Thr Leu Asn Gln Ile Thr Pro Leu Tyr Asn Met Val Arg Asn Tyr
530 535 540
Met Thr Arg Lys Pro Tyr Ser Gln Glu Lys Ile Lys Leu Asn Phe Glu
545 550 555 560
Asn Ser Thr Leu Leu Gly Gly Trp Asp Leu Asn Lys Glu His Asp Asn
565 570 575
Thr Ala Ile Ile Leu Arg Lys Asn Gly Leu Tyr Tyr Leu Ala Ile Met
580 585 590
Lys Lys Ser Ala Asn Lys Ile Phe Asp Lys Asp Lys Leu Asp Asn Ser
595 600 605
Gly Asp Cys Tyr Glu Lys Met Val Tyr Lys Leu Leu Pro Gly Ala Asn
610 615 620
Lys Met Leu Pro Lys Val Phe Phe Ser Lys Ser Arg Ile Asp Glu Phe
625 630 635 640
Lys Pro Ser Glu Asn Ile Ile Glu Asn Tyr Lys Lys Gly Thr His Lys
645 650 655
Lys Gly Ala Asn Phe Asn Leu Ala Asp Cys His Asn Leu Ile Asp Phe
660 665 670
Phe Lys Ser Ser Ile Ser Lys His Glu Asp Trp Ser Lys Phe Asn Phe
675 680 685
His Phe Ser Asp Thr Ser Ser Tyr Glu Asp Leu Ser Asp Phe Tyr Arg
690 695 700
Glu Val Glu Gln Gln Gly Tyr Ser Ile Ser Phe Cys Asp Val Ser Val
705 710 715 720
Glu Tyr Ile Asn Lys Met Val Glu Lys Gly Asp Leu Tyr Leu Phe Gln
725 730 735
Ile Tyr Asn Lys Asp Phe Ser Glu Phe Ser Lys Gly Thr Pro Asn Met
740 745 750
His Thr Leu Tyr Trp Asn Ser Leu Phe Ser Lys Glu Asn Leu Asn Asn
755 760 765
Ile Ile Tyr Lys Leu Asn Gly Gln Ala Glu Ile Phe Phe Arg Lys Lys
770 775 780
Ser Leu Asn Tyr Lys Arg Pro Thr His Pro Ala His Gln Ala Ile Lys
785 790 795 800
Asn Lys Asn Lys Cys Asn Glu Lys Lys Glu Ser Ile Phe Asp Tyr Asp
805 810 815
Leu Val Lys Asp Lys Arg Tyr Thr Val Asp Lys Phe Gln Phe His Val
820 825 830
Pro Ile Thr Met Asn Phe Lys Ser Thr Gly Asn Thr Asn Ile Asn Gln
835 840 845
Gln Val Ile Asp Tyr Leu Arg Thr Glu Asp Asp Thr His Ile Ile Gly
850 855 860
Ile Asp Arg Gly Glu Arg His Leu Leu Tyr Leu Val Val Ile Asp Ser
865 870 875 880
His Gly Lys Ile Val Glu Gln Phe Thr Leu Asn Glu Ile Val Asn Glu
885 890 895
Tyr Gly Gly Asn Ile Tyr Arg Thr Asn Tyr His Asp Leu Leu Asp Thr
900 905 910
Arg Glu Gln Asn Arg Glu Lys Ala Arg Glu Ser Trp Gln Thr Ile Glu
915 920 925
Asn Ile Lys Glu Leu Lys Glu Gly Tyr Ile Ser Gln Val Ile His Lys
930 935 940
Ile Thr Asp Leu Met Gln Lys Tyr His Ala Val Val Val Leu Glu Asp
945 950 955 960
Leu Asn Met Gly Phe Met Arg Gly Arg Gln Lys Val Glu Lys Gln Val
965 970 975
Tyr Gln Lys Phe Glu Glu Met Leu Ile Asn Lys Leu Asn Tyr Leu Val
980 985 990
Asn Lys Lys Ala Asp Gln Asn Ser Ala Gly Gly Leu Leu His Ala Tyr
995 1000 1005
Gln Leu Thr Ser Lys Phe Glu Ser Phe Gln Lys Leu Gly Lys Gln
1010 1015 1020
Ser Gly Phe Leu Phe Tyr Ile Pro Ala Trp Asn Thr Ser Lys Ile
1025 1030 1035
Asp Pro Val Thr Gly Phe Val Asn Leu Phe Asp Thr Arg Tyr Glu
1040 1045 1050
Ser Ile Asp Lys Ala Lys Ala Phe Phe Gly Lys Phe Asp Ser Ile
1055 1060 1065
Arg Tyr Asn Ala Asp Lys Asp Trp Phe Glu Phe Ala Phe Asp Tyr
1070 1075 1080
Asn Asn Phe Thr Thr Lys Ala Glu Gly Thr Arg Thr Asn Trp Thr
1085 1090 1095
Ile Cys Thr Tyr Gly Ser Arg Ile Arg Thr Phe Arg Asn Gln Ala
1100 1105 1110
Lys Asn Ser Gln Trp Asp Asn Glu Glu Ile Asp Leu Thr Lys Ala
1115 1120 1125
Tyr Lys Ala Phe Phe Ala Lys His Gly Ile Asn Ile Tyr Asp Asn
1130 1135 1140
Ile Lys Glu Ala Ile Ala Met Glu Thr Glu Lys Ser Phe Phe Glu
1145 1150 1155
Asp Leu Leu His Leu Leu Lys Leu Thr Leu Gln Met Arg Asn Ser
1160 1165 1170
Ile Thr Gly Thr Thr Thr Asp Tyr Leu Ile Ser Pro Val His Asp
1175 1180 1185
Ser Lys Gly Asn Phe Tyr Asp Ser Arg Ile Cys Asp Asn Ser Leu
1190 1195 1200
Pro Ala Asn Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala Arg Lys
1205 1210 1215
Gly Leu Met Leu Ile Gln Gln Ile Lys Asp Ser Thr Ser Ser Asn
1220 1225 1230
Arg Phe Lys Phe Ser Pro Ile Thr Asn Lys Asp Trp Leu Ile Phe
1235 1240 1245
Ala Gln Glu Lys Pro Tyr Leu Asn Asp
1250 1255
<210> 41
<211> 1262
<212> PRT
<213> 口腔拟杆菌
<220>
<221> MISC_FEATURE
<222> (1)..(1262)
<223> Genbank WP_009217842 Cpf1
<400> 41
Met Arg Lys Phe Asn Glu Phe Val Gly Leu Tyr Pro Ile Ser Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Lys Pro Ile Gly Lys Thr Leu Glu His Ile Gln
20 25 30
Arg Asn Lys Leu Leu Glu His Asp Ala Val Arg Ala Asp Asp Tyr Val
35 40 45
Lys Val Lys Lys Ile Ile Asp Lys Tyr His Lys Cys Leu Ile Asp Glu
50 55 60
Ala Leu Ser Gly Phe Thr Phe Asp Thr Glu Ala Asp Gly Arg Ser Asn
65 70 75 80
Asn Ser Leu Ser Glu Tyr Tyr Leu Tyr Tyr Asn Leu Lys Lys Arg Asn
85 90 95
Glu Gln Glu Gln Lys Thr Phe Lys Thr Ile Gln Asn Asn Leu Arg Lys
100 105 110
Gln Ile Val Asn Lys Leu Thr Gln Ser Glu Lys Tyr Lys Arg Ile Asp
115 120 125
Lys Lys Glu Leu Ile Thr Thr Asp Leu Pro Asp Phe Leu Thr Asn Glu
130 135 140
Ser Glu Lys Glu Leu Val Glu Lys Phe Lys Asn Phe Thr Thr Tyr Phe
145 150 155 160
Thr Glu Phe His Lys Asn Arg Lys Asn Met Tyr Ser Lys Glu Glu Lys
165 170 175
Ser Thr Ala Ile Ala Phe Arg Leu Ile Asn Glu Asn Leu Pro Lys Phe
180 185 190
Val Asp Asn Ile Ala Ala Phe Glu Lys Val Val Ser Ser Pro Leu Ala
195 200 205
Glu Lys Ile Asn Ala Leu Tyr Glu Asp Phe Lys Glu Tyr Leu Asn Val
210 215 220
Glu Glu Ile Ser Arg Val Phe Arg Leu Asp Tyr Tyr Asp Glu Leu Leu
225 230 235 240
Thr Gln Lys Gln Ile Asp Leu Tyr Asn Ala Ile Val Gly Gly Arg Thr
245 250 255
Glu Glu Asp Asn Lys Ile Gln Ile Lys Gly Leu Asn Gln Tyr Ile Asn
260 265 270
Glu Tyr Asn Gln Gln Gln Thr Asp Arg Ser Asn Arg Leu Pro Lys Leu
275 280 285
Lys Pro Leu Tyr Lys Gln Ile Leu Ser Asp Arg Glu Ser Val Ser Trp
290 295 300
Leu Pro Pro Lys Phe Asp Ser Asp Lys Asn Leu Leu Ile Lys Ile Lys
305 310 315 320
Glu Cys Tyr Asp Ala Leu Ser Glu Lys Glu Lys Val Phe Asp Lys Leu
325 330 335
Glu Ser Ile Leu Lys Ser Leu Ser Thr Tyr Asp Leu Ser Lys Ile Tyr
340 345 350
Ile Ser Asn Asp Ser Gln Leu Ser Tyr Ile Ser Gln Lys Met Phe Gly
355 360 365
Arg Trp Asp Ile Ile Ser Lys Ala Ile Arg Glu Asp Cys Ala Lys Arg
370 375 380
Asn Pro Gln Lys Ser Arg Glu Ser Leu Glu Lys Phe Ala Glu Arg Ile
385 390 395 400
Asp Lys Lys Leu Lys Thr Ile Asp Ser Ile Ser Ile Gly Asp Val Asp
405 410 415
Glu Cys Leu Ala Gln Leu Gly Glu Thr Tyr Val Lys Arg Val Glu Asp
420 425 430
Tyr Phe Val Ala Met Gly Glu Ser Glu Ile Asp Asp Glu Gln Thr Asp
435 440 445
Thr Thr Ser Phe Lys Lys Asn Ile Glu Gly Ala Tyr Glu Ser Val Lys
450 455 460
Glu Leu Leu Asn Asn Ala Asp Asn Ile Thr Asp Asn Asn Leu Met Gln
465 470 475 480
Asp Lys Gly Asn Val Glu Lys Ile Lys Thr Leu Leu Asp Ala Ile Lys
485 490 495
Asp Leu Gln Arg Phe Ile Lys Pro Leu Leu Gly Lys Gly Asp Glu Ala
500 505 510
Asp Lys Asp Gly Val Phe Tyr Gly Glu Phe Thr Ser Leu Trp Thr Lys
515 520 525
Leu Asp Gln Val Thr Pro Leu Tyr Asn Met Val Arg Asn Tyr Leu Thr
530 535 540
Ser Lys Pro Tyr Ser Thr Lys Lys Ile Lys Leu Asn Phe Glu Asn Ser
545 550 555 560
Thr Leu Met Asp Gly Trp Asp Leu Asn Lys Glu Pro Asp Asn Thr Thr
565 570 575
Val Ile Phe Cys Lys Asp Gly Leu Tyr Tyr Leu Gly Ile Met Gly Lys
580 585 590
Lys Tyr Asn Arg Val Phe Val Asp Arg Glu Asp Leu Pro His Asp Gly
595 600 605
Glu Cys Tyr Asp Lys Met Glu Tyr Lys Leu Leu Pro Gly Ala Asn Lys
610 615 620
Met Leu Pro Lys Val Phe Phe Ser Glu Thr Gly Ile Gln Arg Phe Leu
625 630 635 640
Pro Ser Glu Glu Leu Leu Gly Lys Tyr Glu Arg Gly Thr His Lys Lys
645 650 655
Gly Ala Gly Phe Asp Leu Gly Asp Cys Arg Ala Leu Ile Asp Phe Phe
660 665 670
Lys Lys Ser Ile Glu Arg His Asp Asp Trp Lys Lys Phe Asp Phe Lys
675 680 685
Phe Ser Asp Thr Ser Thr Tyr Gln Asp Ile Ser Glu Phe Tyr Arg Glu
690 695 700
Val Glu Gln Gln Gly Tyr Lys Met Ser Phe Arg Lys Val Ser Val Asp
705 710 715 720
Tyr Ile Lys Ser Leu Val Glu Glu Gly Lys Leu Tyr Leu Phe Gln Ile
725 730 735
Tyr Asn Lys Asp Phe Ser Ala His Ser Lys Gly Thr Pro Asn Met His
740 745 750
Thr Leu Tyr Trp Lys Met Leu Phe Asp Glu Glu Asn Leu Lys Asp Val
755 760 765
Val Tyr Lys Leu Asn Gly Glu Ala Glu Val Phe Phe Arg Lys Ser Ser
770 775 780
Ile Thr Val Gln Ser Pro Thr His Pro Ala Asn Ser Pro Ile Lys Asn
785 790 795 800
Lys Asn Lys Asp Asn Gln Lys Lys Glu Ser Lys Phe Glu Tyr Asp Leu
805 810 815
Ile Lys Asp Arg Arg Tyr Thr Val Asp Lys Phe Leu Phe His Val Pro
820 825 830
Ile Thr Met Asn Phe Lys Ser Val Gly Gly Ser Asn Ile Asn Gln Leu
835 840 845
Val Lys Arg His Ile Arg Ser Ala Thr Asp Leu His Ile Ile Gly Ile
850 855 860
Asp Arg Gly Glu Arg His Leu Leu Tyr Leu Thr Val Ile Asp Ser Arg
865 870 875 880
Gly Asn Ile Lys Glu Gln Phe Ser Leu Asn Glu Ile Val Asn Glu Tyr
885 890 895
Asn Gly Asn Thr Tyr Arg Thr Asp Tyr His Glu Leu Leu Asp Thr Arg
900 905 910
Glu Gly Glu Arg Thr Glu Ala Arg Arg Asn Trp Gln Thr Ile Gln Asn
915 920 925
Ile Arg Glu Leu Lys Glu Gly Tyr Leu Ser Gln Val Ile His Lys Ile
930 935 940
Ser Glu Leu Ala Ile Lys Tyr Asn Ala Val Ile Val Leu Glu Asp Leu
945 950 955 960
Asn Phe Gly Phe Met Arg Ser Arg Gln Lys Val Glu Lys Gln Val Tyr
965 970 975
Gln Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Leu Val Asp
980 985 990
Lys Lys Lys Pro Val Ala Glu Thr Gly Gly Leu Leu Arg Ala Tyr Gln
995 1000 1005
Leu Thr Gly Glu Phe Glu Ser Phe Lys Thr Leu Gly Lys Gln Ser
1010 1015 1020
Gly Ile Leu Phe Tyr Val Pro Ala Trp Asn Thr Ser Lys Ile Asp
1025 1030 1035
Pro Val Thr Gly Phe Val Asn Leu Phe Asp Thr His Tyr Glu Asn
1040 1045 1050
Ile Glu Lys Ala Lys Val Phe Phe Asp Lys Phe Lys Ser Ile Arg
1055 1060 1065
Tyr Asn Ser Asp Lys Asp Trp Phe Glu Phe Val Val Asp Asp Tyr
1070 1075 1080
Thr Arg Phe Ser Pro Lys Ala Glu Gly Thr Arg Arg Asp Trp Thr
1085 1090 1095
Ile Cys Thr Gln Gly Lys Arg Ile Gln Ile Cys Arg Asn His Gln
1100 1105 1110
Arg Asn Asn Glu Trp Glu Gly Gln Glu Ile Asp Leu Thr Lys Ala
1115 1120 1125
Phe Lys Glu His Phe Glu Ala Tyr Gly Val Asp Ile Ser Lys Asp
1130 1135 1140
Leu Arg Glu Gln Ile Asn Thr Gln Asn Lys Lys Glu Phe Phe Glu
1145 1150 1155
Glu Leu Leu Arg Leu Leu Arg Leu Thr Leu Gln Met Arg Asn Ser
1160 1165 1170
Met Pro Ser Ser Asp Ile Asp Tyr Leu Ile Ser Pro Val Ala Asn
1175 1180 1185
Asp Thr Gly Cys Phe Phe Asp Ser Arg Lys Gln Ala Glu Leu Lys
1190 1195 1200
Glu Asn Ala Val Leu Pro Met Asn Ala Asp Ala Asn Gly Ala Tyr
1205 1210 1215
Asn Ile Ala Arg Lys Gly Leu Leu Ala Ile Arg Lys Met Lys Gln
1220 1225 1230
Glu Glu Asn Asp Ser Ala Lys Ile Ser Leu Ala Ile Ser Asn Lys
1235 1240 1245
Glu Trp Leu Lys Phe Ala Gln Thr Lys Pro Tyr Leu Glu Asp
1250 1255 1260
<210> 42
<211> 1318
<212> PRT
<213> 嗜鳃黄杆菌
<220>
<221> MISC_FEATURE
<222> (1)..(1318)
<223> Genbank WP_014085038 Cpf1
<400> 42
Met Thr Asn Lys Phe Thr Asn Gln Tyr Ser Leu Ser Lys Thr Leu Arg
1 5 10 15
Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Glu Phe Ile Gln Glu Lys
20 25 30
Gly Leu Leu Ser Gln Asp Lys Gln Arg Ala Glu Ser Tyr Gln Glu Met
35 40 45
Lys Lys Thr Ile Asp Lys Phe His Lys Tyr Phe Ile Asp Leu Ala Leu
50 55 60
Ser Asn Ala Lys Leu Thr His Leu Glu Thr Tyr Leu Glu Leu Tyr Asn
65 70 75 80
Lys Ser Ala Glu Thr Lys Lys Glu Gln Lys Phe Lys Asp Asp Leu Lys
85 90 95
Lys Val Gln Asp Asn Leu Arg Lys Glu Ile Val Lys Ser Phe Ser Asp
100 105 110
Gly Asp Ala Lys Ser Ile Phe Ala Ile Leu Asp Lys Lys Glu Leu Ile
115 120 125
Thr Val Glu Leu Glu Lys Trp Phe Glu Asn Asn Glu Gln Lys Asp Ile
130 135 140
Tyr Phe Asp Glu Lys Phe Lys Thr Phe Thr Thr Tyr Phe Thr Gly Phe
145 150 155 160
His Gln Asn Arg Lys Asn Met Tyr Ser Val Glu Pro Asn Ser Thr Ala
165 170 175
Ile Ala Tyr Arg Leu Ile His Glu Asn Leu Pro Lys Phe Leu Glu Asn
180 185 190
Ala Lys Ala Phe Glu Lys Ile Lys Gln Val Glu Ser Leu Gln Val Asn
195 200 205
Phe Arg Glu Leu Met Gly Glu Phe Gly Asp Glu Gly Leu Ile Phe Val
210 215 220
Asn Glu Leu Glu Glu Met Phe Gln Ile Asn Tyr Tyr Asn Asp Val Leu
225 230 235 240
Ser Gln Asn Gly Ile Thr Ile Tyr Asn Ser Ile Ile Ser Gly Phe Thr
245 250 255
Lys Asn Asp Ile Lys Tyr Lys Gly Leu Asn Glu Tyr Ile Asn Asn Tyr
260 265 270
Asn Gln Thr Lys Asp Lys Lys Asp Arg Leu Pro Lys Leu Lys Gln Leu
275 280 285
Tyr Lys Gln Ile Leu Ser Asp Arg Ile Ser Leu Ser Phe Leu Pro Asp
290 295 300
Ala Phe Thr Asp Gly Lys Gln Val Leu Lys Ala Ile Phe Asp Phe Tyr
305 310 315 320
Lys Ile Asn Leu Leu Ser Tyr Thr Ile Glu Gly Gln Glu Glu Ser Gln
325 330 335
Asn Leu Leu Leu Leu Ile Arg Gln Thr Ile Glu Asn Leu Ser Ser Phe
340 345 350
Asp Thr Gln Lys Ile Tyr Leu Lys Asn Asp Thr His Leu Thr Thr Ile
355 360 365
Ser Gln Gln Val Phe Gly Asp Phe Ser Val Phe Ser Thr Ala Leu Asn
370 375 380
Tyr Trp Tyr Glu Thr Lys Val Asn Pro Lys Phe Glu Thr Glu Tyr Ser
385 390 395 400
Lys Ala Asn Glu Lys Lys Arg Glu Ile Leu Asp Lys Ala Lys Ala Val
405 410 415
Phe Thr Lys Gln Asp Tyr Phe Ser Ile Ala Phe Leu Gln Glu Val Leu
420 425 430
Ser Glu Tyr Ile Leu Thr Leu Asp His Thr Ser Asp Ile Val Lys Lys
435 440 445
His Ser Ser Asn Cys Ile Ala Asp Tyr Phe Lys Asn His Phe Val Ala
450 455 460
Lys Lys Glu Asn Glu Thr Asp Lys Thr Phe Asp Phe Ile Ala Asn Ile
465 470 475 480
Thr Ala Lys Tyr Gln Cys Ile Gln Gly Ile Leu Glu Asn Ala Asp Gln
485 490 495
Tyr Glu Asp Glu Leu Lys Gln Asp Gln Lys Leu Ile Asp Asn Leu Lys
500 505 510
Phe Phe Leu Asp Ala Ile Leu Glu Leu Leu His Phe Ile Lys Pro Leu
515 520 525
His Leu Lys Ser Glu Ser Ile Thr Glu Lys Asp Thr Ala Phe Tyr Asp
530 535 540
Val Phe Glu Asn Tyr Tyr Glu Ala Leu Ser Leu Leu Thr Pro Leu Tyr
545 550 555 560
Asn Met Val Arg Asn Tyr Val Thr Gln Lys Pro Tyr Ser Thr Glu Lys
565 570 575
Ile Lys Leu Asn Phe Glu Asn Ala Gln Leu Leu Asn Gly Trp Asp Ala
580 585 590
Asn Lys Glu Gly Asp Tyr Leu Thr Thr Ile Leu Lys Lys Asp Gly Asn
595 600 605
Tyr Phe Leu Ala Ile Met Asp Lys Lys His Asn Lys Ala Phe Gln Lys
610 615 620
Phe Pro Glu Gly Lys Glu Asn Tyr Glu Lys Met Val Tyr Lys Leu Leu
625 630 635 640
Pro Gly Val Asn Lys Met Leu Pro Lys Val Phe Phe Ser Asn Lys Asn
645 650 655
Ile Ala Tyr Phe Asn Pro Ser Lys Glu Leu Leu Glu Asn Tyr Lys Lys
660 665 670
Glu Thr His Lys Lys Gly Asp Thr Phe Asn Leu Glu His Cys His Thr
675 680 685
Leu Ile Asp Phe Phe Lys Asp Ser Leu Asn Lys His Glu Asp Trp Lys
690 695 700
Tyr Phe Asp Phe Gln Phe Ser Glu Thr Lys Ser Tyr Gln Asp Leu Ser
705 710 715 720
Gly Phe Tyr Arg Glu Val Glu His Gln Gly Tyr Lys Ile Asn Phe Lys
725 730 735
Asn Ile Asp Ser Glu Tyr Ile Asp Gly Leu Val Asn Glu Gly Lys Leu
740 745 750
Phe Leu Phe Gln Ile Tyr Ser Lys Asp Phe Ser Pro Phe Ser Lys Gly
755 760 765
Lys Pro Asn Met His Thr Leu Tyr Trp Lys Ala Leu Phe Glu Glu Gln
770 775 780
Asn Leu Gln Asn Val Ile Tyr Lys Leu Asn Gly Gln Ala Glu Ile Phe
785 790 795 800
Phe Arg Lys Ala Ser Ile Lys Pro Lys Asn Ile Ile Leu His Lys Lys
805 810 815
Lys Ile Lys Ile Ala Lys Lys His Phe Ile Asp Lys Lys Thr Lys Thr
820 825 830
Ser Glu Ile Val Pro Val Gln Thr Ile Lys Asn Leu Asn Met Tyr Tyr
835 840 845
Gln Gly Lys Ile Ser Glu Lys Glu Leu Thr Gln Asp Asp Leu Arg Tyr
850 855 860
Ile Asp Asn Phe Ser Ile Phe Asn Glu Lys Asn Lys Thr Ile Asp Ile
865 870 875 880
Ile Lys Asp Lys Arg Phe Thr Val Asp Lys Phe Gln Phe His Val Pro
885 890 895
Ile Thr Met Asn Phe Lys Ala Thr Gly Gly Ser Tyr Ile Asn Gln Thr
900 905 910
Val Leu Glu Tyr Leu Gln Asn Asn Pro Glu Val Lys Ile Ile Gly Leu
915 920 925
Asp Arg Gly Glu Arg His Leu Val Tyr Leu Thr Leu Ile Asp Gln Gln
930 935 940
Gly Asn Ile Leu Lys Gln Glu Ser Leu Asn Thr Ile Thr Asp Ser Lys
945 950 955 960
Ile Ser Thr Pro Tyr His Lys Leu Leu Asp Asn Lys Glu Asn Glu Arg
965 970 975
Asp Leu Ala Arg Lys Asn Trp Gly Thr Val Glu Asn Ile Lys Glu Leu
980 985 990
Lys Glu Gly Tyr Ile Ser Gln Val Val His Lys Ile Ala Thr Leu Met
995 1000 1005
Leu Glu Glu Asn Ala Ile Val Val Met Glu Asp Leu Asn Phe Gly
1010 1015 1020
Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Ile Tyr Gln Lys
1025 1030 1035
Leu Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Leu Val Leu Lys
1040 1045 1050
Asp Lys Gln Pro Gln Glu Leu Gly Gly Leu Tyr Asn Ala Leu Gln
1055 1060 1065
Leu Thr Asn Lys Phe Glu Ser Phe Gln Lys Met Gly Lys Gln Ser
1070 1075 1080
Gly Phe Leu Phe Tyr Val Pro Ala Trp Asn Thr Ser Lys Ile Asp
1085 1090 1095
Pro Thr Thr Gly Phe Val Asn Tyr Phe Tyr Thr Lys Tyr Glu Asn
1100 1105 1110
Val Asp Lys Ala Lys Ala Phe Phe Glu Lys Phe Glu Ala Ile Arg
1115 1120 1125
Phe Asn Ala Glu Lys Lys Tyr Phe Glu Phe Glu Val Lys Lys Tyr
1130 1135 1140
Ser Asp Phe Asn Pro Lys Ala Glu Gly Thr Gln Gln Ala Trp Thr
1145 1150 1155
Ile Cys Thr Tyr Gly Glu Arg Ile Glu Thr Lys Arg Gln Lys Asp
1160 1165 1170
Gln Asn Asn Lys Phe Val Ser Thr Pro Ile Asn Leu Thr Glu Lys
1175 1180 1185
Ile Glu Asp Phe Leu Gly Lys Asn Gln Ile Val Tyr Gly Asp Gly
1190 1195 1200
Asn Cys Ile Lys Ser Gln Ile Ala Ser Lys Asp Asp Lys Ala Phe
1205 1210 1215
Phe Glu Thr Leu Leu Tyr Trp Phe Lys Met Thr Leu Gln Met Arg
1220 1225 1230
Asn Ser Glu Thr Arg Thr Asp Ile Asp Tyr Leu Ile Ser Pro Val
1235 1240 1245
Met Asn Asp Asn Gly Thr Phe Tyr Asn Ser Arg Asp Tyr Glu Lys
1250 1255 1260
Leu Glu Asn Pro Thr Leu Pro Lys Asp Ala Asp Ala Asn Gly Ala
1265 1270 1275
Tyr His Ile Ala Lys Lys Gly Leu Met Leu Leu Asn Lys Ile Asp
1280 1285 1290
Gln Ala Asp Leu Thr Lys Lys Val Asp Leu Ser Ile Ser Asn Arg
1295 1300 1305
Asp Trp Leu Gln Phe Val Gln Lys Asn Lys
1310 1315
<210> 43
<211> 1230
<212> PRT
<213> 毛螺菌科细菌
<220>
<221> MISC_FEATURE
<222> (1)..(1230)
<223> Genbank WP_016301126 Cpf1
<400> 43
Met His Glu Asn Asn Gly Lys Ile Ala Asp Asn Phe Ile Gly Ile Tyr
1 5 10 15
Pro Val Ser Lys Thr Leu Arg Phe Glu Leu Lys Pro Val Gly Lys Thr
20 25 30
Gln Glu Tyr Ile Glu Lys His Gly Ile Leu Asp Glu Asp Leu Lys Arg
35 40 45
Ala Gly Asp Tyr Lys Ser Val Lys Lys Ile Ile Asp Ala Tyr His Lys
50 55 60
Tyr Phe Ile Asp Glu Ala Leu Asn Gly Ile Gln Leu Asp Gly Leu Lys
65 70 75 80
Asn Tyr Tyr Glu Leu Tyr Glu Lys Lys Arg Asp Asn Asn Glu Glu Lys
85 90 95
Glu Phe Gln Lys Ile Gln Met Ser Leu Arg Lys Gln Ile Val Lys Arg
100 105 110
Phe Ser Glu His Pro Gln Tyr Lys Tyr Leu Phe Lys Lys Glu Leu Ile
115 120 125
Lys Asn Val Leu Pro Glu Phe Thr Lys Asp Asn Ala Glu Glu Gln Thr
130 135 140
Leu Val Lys Ser Phe Gln Glu Phe Thr Thr Tyr Phe Glu Gly Phe His
145 150 155 160
Gln Asn Arg Lys Asn Met Tyr Ser Asp Glu Glu Lys Ser Thr Ala Ile
165 170 175
Ala Tyr Arg Val Val His Gln Asn Leu Pro Lys Tyr Ile Asp Asn Met
180 185 190
Arg Ile Phe Ser Met Ile Leu Asn Thr Asp Ile Arg Ser Asp Leu Thr
195 200 205
Glu Leu Phe Asn Asn Leu Lys Thr Lys Met Asp Ile Thr Ile Val Glu
210 215 220
Glu Tyr Phe Ala Ile Asp Gly Phe Asn Lys Val Val Asn Gln Lys Gly
225 230 235 240
Ile Asp Val Tyr Asn Thr Ile Leu Gly Ala Phe Ser Thr Asp Asp Asn
245 250 255
Thr Lys Ile Lys Gly Leu Asn Glu Tyr Ile Asn Leu Tyr Asn Gln Lys
260 265 270
Asn Lys Ala Lys Leu Pro Lys Leu Lys Pro Leu Phe Lys Gln Ile Leu
275 280 285
Ser Asp Arg Asp Lys Ile Ser Phe Ile Pro Glu Gln Phe Asp Ser Asp
290 295 300
Thr Glu Val Leu Glu Ala Val Asp Met Phe Tyr Asn Arg Leu Leu Gln
305 310 315 320
Phe Val Ile Glu Asn Glu Gly Gln Ile Thr Ile Ser Lys Leu Leu Thr
325 330 335
Asn Phe Ser Ala Tyr Asp Leu Asn Lys Ile Tyr Val Lys Asn Asp Thr
340 345 350
Thr Ile Ser Ala Ile Ser Asn Asp Leu Phe Asp Asp Trp Ser Tyr Ile
355 360 365
Ser Lys Ala Val Arg Glu Asn Tyr Asp Ser Glu Asn Val Asp Lys Asn
370 375 380
Lys Arg Ala Ala Ala Tyr Glu Glu Lys Lys Glu Lys Ala Leu Ser Lys
385 390 395 400
Ile Lys Met Tyr Ser Ile Glu Glu Leu Asn Phe Phe Val Lys Lys Tyr
405 410 415
Ser Cys Asn Glu Cys His Ile Glu Gly Tyr Phe Glu Arg Arg Ile Leu
420 425 430
Glu Ile Leu Asp Lys Met Arg Tyr Ala Tyr Glu Ser Cys Lys Ile Leu
435 440 445
His Asp Lys Gly Leu Ile Asn Asn Ile Ser Leu Cys Gln Asp Arg Gln
450 455 460
Ala Ile Ser Glu Leu Lys Asp Phe Leu Asp Ser Ile Lys Glu Val Gln
465 470 475 480
Trp Leu Leu Lys Pro Leu Met Ile Gly Gln Glu Gln Ala Asp Lys Glu
485 490 495
Glu Ala Phe Tyr Thr Glu Leu Leu Arg Ile Trp Glu Glu Leu Glu Pro
500 505 510
Ile Thr Leu Leu Tyr Asn Lys Val Arg Asn Tyr Val Thr Lys Lys Pro
515 520 525
Tyr Thr Leu Glu Lys Val Lys Leu Asn Phe Tyr Lys Ser Thr Leu Leu
530 535 540
Asp Gly Trp Asp Lys Asn Lys Glu Lys Asp Asn Leu Gly Ile Ile Leu
545 550 555 560
Leu Lys Asp Gly Gln Tyr Tyr Leu Gly Ile Met Asn Arg Arg Asn Asn
565 570 575
Lys Ile Ala Asp Asp Ala Pro Leu Ala Lys Thr Asp Asn Val Tyr Arg
580 585 590
Lys Met Glu Tyr Lys Leu Leu Thr Lys Val Ser Ala Asn Leu Pro Arg
595 600 605
Ile Phe Leu Lys Asp Lys Tyr Asn Pro Ser Glu Glu Met Leu Glu Lys
610 615 620
Tyr Glu Lys Gly Thr His Leu Lys Gly Glu Asn Phe Cys Ile Asp Asp
625 630 635 640
Cys Arg Glu Leu Ile Asp Phe Phe Lys Lys Gly Ile Lys Gln Tyr Glu
645 650 655
Asp Trp Gly Gln Phe Asp Phe Lys Phe Ser Asp Thr Glu Ser Tyr Asp
660 665 670
Asp Ile Ser Ala Phe Tyr Lys Glu Val Glu His Gln Gly Tyr Lys Ile
675 680 685
Thr Phe Arg Asp Ile Asp Glu Thr Tyr Ile Asp Ser Leu Val Asn Glu
690 695 700
Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Pro Tyr
705 710 715 720
Ser Lys Gly Thr Lys Asn Leu His Thr Leu Tyr Trp Glu Met Leu Phe
725 730 735
Ser Gln Gln Asn Leu Gln Asn Ile Val Tyr Lys Leu Asn Gly Asn Ala
740 745 750
Glu Ile Phe Tyr Arg Lys Ala Ser Ile Asn Gln Lys Asp Val Val Val
755 760 765
His Lys Ala Asp Leu Pro Ile Lys Asn Lys Asp Pro Gln Asn Ser Lys
770 775 780
Lys Glu Ser Met Phe Asp Tyr Asp Ile Ile Lys Asp Lys Arg Phe Thr
785 790 795 800
Cys Asp Lys Tyr Gln Phe His Val Pro Ile Thr Met Asn Phe Lys Ala
805 810 815
Leu Gly Glu Asn His Phe Asn Arg Lys Val Asn Arg Leu Ile His Asp
820 825 830
Ala Glu Asn Met His Ile Ile Gly Ile Asp Arg Gly Glu Arg Asn Leu
835 840 845
Ile Tyr Leu Cys Met Ile Asp Met Lys Gly Asn Ile Val Lys Gln Ile
850 855 860
Ser Leu Asn Glu Ile Ile Ser Tyr Asp Lys Asn Lys Leu Glu His Lys
865 870 875 880
Arg Asn Tyr His Gln Leu Leu Lys Thr Arg Glu Asp Glu Asn Lys Ser
885 890 895
Ala Arg Gln Ser Trp Gln Thr Ile His Thr Ile Lys Glu Leu Lys Glu
900 905 910
Gly Tyr Leu Ser Gln Val Ile His Val Ile Thr Asp Leu Met Val Glu
915 920 925
Tyr Asn Ala Ile Val Val Leu Glu Asp Leu Asn Phe Gly Phe Lys Gln
930 935 940
Gly Arg Gln Lys Phe Glu Arg Gln Val Tyr Gln Lys Phe Glu Lys Met
945 950 955 960
Leu Ile Asp Lys Leu Asn Tyr Leu Val Asp Lys Ser Lys Gly Met Asp
965 970 975
Glu Asp Gly Gly Leu Leu His Ala Tyr Gln Leu Thr Asp Glu Phe Lys
980 985 990
Ser Phe Lys Gln Leu Gly Lys Gln Ser Gly Phe Leu Tyr Tyr Ile Pro
995 1000 1005
Ala Trp Asn Thr Ser Lys Leu Asp Pro Thr Thr Gly Phe Val Asn
1010 1015 1020
Leu Phe Tyr Thr Lys Tyr Glu Ser Val Glu Lys Ser Lys Glu Phe
1025 1030 1035
Ile Asn Asn Phe Thr Ser Ile Leu Tyr Asn Gln Glu Arg Glu Tyr
1040 1045 1050
Phe Glu Phe Leu Phe Asp Tyr Ser Ala Phe Thr Ser Lys Ala Glu
1055 1060 1065
Gly Ser Arg Leu Lys Trp Thr Val Cys Ser Lys Gly Glu Arg Val
1070 1075 1080
Glu Thr Tyr Arg Asn Pro Lys Lys Asn Asn Glu Trp Asp Thr Gln
1085 1090 1095
Lys Ile Asp Leu Thr Phe Glu Leu Lys Lys Leu Phe Asn Asp Tyr
1100 1105 1110
Ser Ile Ser Leu Leu Asp Gly Asp Leu Arg Glu Gln Met Gly Lys
1115 1120 1125
Ile Asp Lys Ala Asp Phe Tyr Lys Lys Phe Met Lys Leu Phe Ala
1130 1135 1140
Leu Ile Val Gln Met Arg Asn Ser Asp Glu Arg Glu Asp Lys Leu
1145 1150 1155
Ile Ser Pro Val Leu Asn Lys Tyr Gly Ala Phe Phe Glu Thr Gly
1160 1165 1170
Lys Asn Glu Arg Met Pro Leu Asp Ala Asp Ala Asn Gly Ala Tyr
1175 1180 1185
Asn Ile Ala Arg Lys Gly Leu Trp Ile Ile Glu Lys Ile Lys Asn
1190 1195 1200
Thr Asp Val Glu Gln Leu Asp Lys Val Lys Leu Thr Ile Ser Asn
1205 1210 1215
Lys Glu Trp Leu Gln Tyr Ala Gln Glu His Ile Leu
1220 1225 1230
<210> 44
<211> 1246
<212> PRT
<213> 猕猴卟啉单胞菌
<220>
<221> MISC_FEATURE
<222> (1)..(1246)
<223> Genbank WP_018359861 Cpf1
<400> 44
Met Lys Thr Gln His Phe Phe Glu Asp Phe Thr Ser Leu Tyr Ser Leu
1 5 10 15
Ser Lys Thr Ile Arg Phe Glu Leu Lys Pro Ile Gly Lys Thr Leu Glu
20 25 30
Asn Ile Lys Lys Asn Gly Leu Ile Arg Arg Asp Glu Gln Arg Leu Asp
35 40 45
Asp Tyr Glu Lys Leu Lys Lys Val Ile Asp Glu Tyr His Glu Asp Phe
50 55 60
Ile Ala Asn Ile Leu Ser Ser Phe Ser Phe Ser Glu Glu Ile Leu Gln
65 70 75 80
Ser Tyr Ile Gln Asn Leu Ser Glu Ser Glu Ala Arg Ala Lys Ile Glu
85 90 95
Lys Thr Met Arg Asp Thr Leu Ala Lys Ala Phe Ser Glu Asp Glu Arg
100 105 110
Tyr Lys Ser Ile Phe Lys Lys Glu Leu Val Lys Lys Asp Ile Pro Val
115 120 125
Trp Cys Pro Ala Tyr Lys Ser Leu Cys Lys Lys Phe Asp Asn Phe Thr
130 135 140
Thr Ser Leu Val Pro Phe His Glu Asn Arg Lys Asn Leu Tyr Thr Ser
145 150 155 160
Asn Glu Ile Thr Ala Ser Ile Pro Tyr Arg Ile Val His Val Asn Leu
165 170 175
Pro Lys Phe Ile Gln Asn Ile Glu Ala Leu Cys Glu Leu Gln Lys Lys
180 185 190
Met Gly Ala Asp Leu Tyr Leu Glu Met Met Glu Asn Leu Arg Asn Val
195 200 205
Trp Pro Ser Phe Val Lys Thr Pro Asp Asp Leu Cys Asn Leu Lys Thr
210 215 220
Tyr Asn His Leu Met Val Gln Ser Ser Ile Ser Glu Tyr Asn Arg Phe
225 230 235 240
Val Gly Gly Tyr Ser Thr Glu Asp Gly Thr Lys His Gln Gly Ile Asn
245 250 255
Glu Trp Ile Asn Ile Tyr Arg Gln Arg Asn Lys Glu Met Arg Leu Pro
260 265 270
Gly Leu Val Phe Leu His Lys Gln Ile Leu Ala Lys Val Asp Ser Ser
275 280 285
Ser Phe Ile Ser Asp Thr Leu Glu Asn Asp Asp Gln Val Phe Cys Val
290 295 300
Leu Arg Gln Phe Arg Lys Leu Phe Trp Asn Thr Val Ser Ser Lys Glu
305 310 315 320
Asp Asp Ala Ala Ser Leu Lys Asp Leu Phe Cys Gly Leu Ser Gly Tyr
325 330 335
Asp Pro Glu Ala Ile Tyr Val Ser Asp Ala His Leu Ala Thr Ile Ser
340 345 350
Lys Asn Ile Phe Asp Arg Trp Asn Tyr Ile Ser Asp Ala Ile Arg Arg
355 360 365
Lys Thr Glu Val Leu Met Pro Arg Lys Lys Glu Ser Val Glu Arg Tyr
370 375 380
Ala Glu Lys Ile Ser Lys Gln Ile Lys Lys Arg Gln Ser Tyr Ser Leu
385 390 395 400
Ala Glu Leu Asp Asp Leu Leu Ala His Tyr Ser Glu Glu Ser Leu Pro
405 410 415
Ala Gly Phe Ser Leu Leu Ser Tyr Phe Thr Ser Leu Gly Gly Gln Lys
420 425 430
Tyr Leu Val Ser Asp Gly Glu Val Ile Leu Tyr Glu Glu Gly Ser Asn
435 440 445
Ile Trp Asp Glu Val Leu Ile Ala Phe Arg Asp Leu Gln Val Ile Leu
450 455 460
Asp Lys Asp Phe Thr Glu Lys Lys Leu Gly Lys Asp Glu Glu Ala Val
465 470 475 480
Ser Val Ile Lys Lys Ala Leu Asp Ser Ala Leu Arg Leu Arg Lys Phe
485 490 495
Phe Asp Leu Leu Ser Gly Thr Gly Ala Glu Ile Arg Arg Asp Ser Ser
500 505 510
Phe Tyr Ala Leu Tyr Thr Asp Arg Met Asp Lys Leu Lys Gly Leu Leu
515 520 525
Lys Met Tyr Asp Lys Val Arg Asn Tyr Leu Thr Lys Lys Pro Tyr Ser
530 535 540
Ile Glu Lys Phe Lys Leu His Phe Asp Asn Pro Ser Leu Leu Ser Gly
545 550 555 560
Trp Asp Lys Asn Lys Glu Leu Asn Asn Leu Ser Val Ile Phe Arg Gln
565 570 575
Asn Gly Tyr Tyr Tyr Leu Gly Ile Met Thr Pro Lys Gly Lys Asn Leu
580 585 590
Phe Lys Thr Leu Pro Lys Leu Gly Ala Glu Glu Met Phe Tyr Glu Lys
595 600 605
Met Glu Tyr Lys Gln Ile Ala Glu Pro Met Leu Met Leu Pro Lys Val
610 615 620
Phe Phe Pro Lys Lys Thr Lys Pro Ala Phe Ala Pro Asp Gln Ser Val
625 630 635 640
Val Asp Ile Tyr Asn Lys Lys Thr Phe Lys Thr Gly Gln Lys Gly Phe
645 650 655
Asn Lys Lys Asp Leu Tyr Arg Leu Ile Asp Phe Tyr Lys Glu Ala Leu
660 665 670
Thr Val His Glu Trp Lys Leu Phe Asn Phe Ser Phe Ser Pro Thr Glu
675 680 685
Gln Tyr Arg Asn Ile Gly Glu Phe Phe Asp Glu Val Arg Glu Gln Ala
690 695 700
Tyr Lys Val Ser Met Val Asn Val Pro Ala Ser Tyr Ile Asp Glu Ala
705 710 715 720
Val Glu Asn Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe
725 730 735
Ser Pro Tyr Ser Lys Gly Ile Pro Asn Leu His Thr Leu Tyr Trp Lys
740 745 750
Ala Leu Phe Ser Glu Gln Asn Gln Ser Arg Val Tyr Lys Leu Cys Gly
755 760 765
Gly Gly Glu Leu Phe Tyr Arg Lys Ala Ser Leu His Met Gln Asp Thr
770 775 780
Thr Val His Pro Lys Gly Ile Ser Ile His Lys Lys Asn Leu Asn Lys
785 790 795 800
Lys Gly Glu Thr Ser Leu Phe Asn Tyr Asp Leu Val Lys Asp Lys Arg
805 810 815
Phe Thr Glu Asp Lys Phe Phe Phe His Val Pro Ile Ser Ile Asn Tyr
820 825 830
Lys Asn Lys Lys Ile Thr Asn Val Asn Gln Met Val Arg Asp Tyr Ile
835 840 845
Ala Gln Asn Asp Asp Leu Gln Ile Ile Gly Ile Asp Arg Gly Glu Arg
850 855 860
Asn Leu Leu Tyr Ile Ser Arg Ile Asp Thr Arg Gly Asn Leu Leu Glu
865 870 875 880
Gln Phe Ser Leu Asn Val Ile Glu Ser Asp Lys Gly Asp Leu Arg Thr
885 890 895
Asp Tyr Gln Lys Ile Leu Gly Asp Arg Glu Gln Glu Arg Leu Arg Arg
900 905 910
Arg Gln Glu Trp Lys Ser Ile Glu Ser Ile Lys Asp Leu Lys Asp Gly
915 920 925
Tyr Met Ser Gln Val Val His Lys Ile Cys Asn Met Val Val Glu His
930 935 940
Lys Ala Ile Val Val Leu Glu Asn Leu Asn Leu Ser Phe Met Lys Gly
945 950 955 960
Arg Lys Lys Val Glu Lys Ser Val Tyr Glu Lys Phe Glu Arg Met Leu
965 970 975
Val Asp Lys Leu Asn Tyr Leu Val Val Asp Lys Lys Asn Leu Ser Asn
980 985 990
Glu Pro Gly Gly Leu Tyr Ala Ala Tyr Gln Leu Thr Asn Pro Leu Phe
995 1000 1005
Ser Phe Glu Glu Leu His Arg Tyr Pro Gln Ser Gly Ile Leu Phe
1010 1015 1020
Phe Val Asp Pro Trp Asn Thr Ser Leu Thr Asp Pro Ser Thr Gly
1025 1030 1035
Phe Val Asn Leu Leu Gly Arg Ile Asn Tyr Thr Asn Val Gly Asp
1040 1045 1050
Ala Arg Lys Phe Phe Asp Arg Phe Asn Ala Ile Arg Tyr Asp Gly
1055 1060 1065
Lys Gly Asn Ile Leu Phe Asp Leu Asp Leu Ser Arg Phe Asp Val
1070 1075 1080
Arg Val Glu Thr Gln Arg Lys Leu Trp Thr Leu Thr Thr Phe Gly
1085 1090 1095
Ser Arg Ile Ala Lys Ser Lys Lys Ser Gly Lys Trp Met Val Glu
1100 1105 1110
Arg Ile Glu Asn Leu Ser Leu Cys Phe Leu Glu Leu Phe Glu Gln
1115 1120 1125
Phe Asn Ile Gly Tyr Arg Val Glu Lys Asp Leu Lys Lys Ala Ile
1130 1135 1140
Leu Ser Gln Asp Arg Lys Glu Phe Tyr Val Arg Leu Ile Tyr Leu
1145 1150 1155
Phe Asn Leu Met Met Gln Ile Arg Asn Ser Asp Gly Glu Glu Asp
1160 1165 1170
Tyr Ile Leu Ser Pro Ala Leu Asn Glu Lys Asn Leu Gln Phe Asp
1175 1180 1185
Ser Arg Leu Ile Glu Ala Lys Asp Leu Pro Val Asp Ala Asp Ala
1190 1195 1200
Asn Gly Ala Tyr Asn Val Ala Arg Lys Gly Leu Met Val Val Gln
1205 1210 1215
Arg Ile Lys Arg Gly Asp His Glu Ser Ile His Arg Ile Gly Arg
1220 1225 1230
Ala Gln Trp Leu Arg Tyr Val Gln Glu Gly Ile Val Glu
1235 1240 1245
<210> 45
<211> 1259
<212> PRT
<213> 琼斯氏共生菌
<220>
<221> MISC_FEATURE
<222> (1)..(1259)
<223> Genbank WP_037975888 V型CV CRISPR关联蛋白Cpf1
<400> 45
Met Ala Asn Ser Leu Lys Asp Phe Thr Asn Ile Tyr Gln Leu Ser Lys
1 5 10 15
Thr Leu Arg Phe Glu Leu Lys Pro Ile Gly Lys Thr Glu Glu His Ile
20 25 30
Asn Arg Lys Leu Ile Ile Met His Asp Glu Lys Arg Gly Glu Asp Tyr
35 40 45
Lys Ser Val Thr Lys Leu Ile Asp Asp Tyr His Arg Lys Phe Ile His
50 55 60
Glu Thr Leu Asp Pro Ala His Phe Asp Trp Asn Pro Leu Ala Glu Ala
65 70 75 80
Leu Ile Gln Ser Gly Ser Lys Asn Asn Lys Ala Leu Pro Ala Glu Gln
85 90 95
Lys Glu Met Arg Glu Lys Ile Ile Ser Met Phe Thr Ser Gln Ala Val
100 105 110
Tyr Lys Lys Leu Phe Lys Lys Glu Leu Phe Ser Glu Leu Leu Pro Glu
115 120 125
Met Ile Lys Ser Glu Leu Val Ser Asp Leu Glu Lys Gln Ala Gln Leu
130 135 140
Asp Ala Val Lys Ser Phe Asp Lys Phe Ser Thr Tyr Phe Thr Gly Phe
145 150 155 160
His Glu Asn Arg Lys Asn Ile Tyr Ser Lys Lys Asp Thr Ser Thr Ser
165 170 175
Ile Ala Phe Arg Ile Val His Gln Asn Phe Pro Lys Phe Leu Ala Asn
180 185 190
Val Arg Ala Tyr Thr Leu Ile Lys Glu Arg Ala Pro Glu Val Ile Asp
195 200 205
Lys Ala Gln Lys Glu Leu Ser Gly Ile Leu Gly Gly Lys Thr Leu Asp
210 215 220
Asp Ile Phe Ser Ile Glu Ser Phe Asn Asn Val Leu Thr Gln Asp Lys
225 230 235 240
Ile Asp Tyr Tyr Asn Gln Ile Ile Gly Gly Val Ser Gly Lys Ala Gly
245 250 255
Asp Lys Lys Leu Arg Gly Val Asn Glu Phe Ser Asn Leu Tyr Arg Gln
260 265 270
Gln His Pro Glu Val Ala Ser Leu Arg Ile Lys Met Val Pro Leu Tyr
275 280 285
Lys Gln Ile Leu Ser Asp Arg Thr Thr Leu Ser Phe Val Pro Glu Ala
290 295 300
Leu Lys Asp Asp Glu Gln Ala Ile Asn Ala Val Asp Gly Leu Arg Ser
305 310 315 320
Glu Leu Glu Arg Asn Asp Ile Phe Asn Arg Ile Lys Arg Leu Phe Gly
325 330 335
Lys Asn Asn Leu Tyr Ser Leu Asp Lys Ile Trp Ile Lys Asn Ser Ser
340 345 350
Ile Ser Ala Phe Ser Asn Glu Leu Phe Lys Asn Trp Ser Phe Ile Glu
355 360 365
Asp Ala Leu Lys Glu Phe Lys Glu Asn Glu Phe Asn Gly Ala Arg Ser
370 375 380
Ala Gly Lys Lys Ala Glu Lys Trp Leu Lys Ser Lys Tyr Phe Ser Phe
385 390 395 400
Ala Asp Ile Asp Ala Ala Val Lys Ser Tyr Ser Glu Gln Val Ser Ala
405 410 415
Asp Ile Ser Ser Ala Pro Ser Ala Ser Tyr Phe Ala Lys Phe Thr Asn
420 425 430
Leu Ile Glu Thr Ala Ala Glu Asn Gly Arg Lys Phe Ser Tyr Phe Ala
435 440 445
Ala Glu Ser Lys Ala Phe Arg Gly Asp Asp Gly Lys Thr Glu Ile Ile
450 455 460
Lys Ala Tyr Leu Asp Ser Leu Asn Asp Ile Leu His Cys Leu Lys Pro
465 470 475 480
Phe Glu Thr Glu Asp Ile Ser Asp Ile Asp Thr Glu Phe Tyr Ser Ala
485 490 495
Phe Ala Glu Ile Tyr Asp Ser Val Lys Asp Val Ile Pro Val Tyr Asn
500 505 510
Ala Val Arg Asn Tyr Thr Thr Gln Lys Pro Phe Ser Thr Glu Lys Phe
515 520 525
Lys Leu Asn Phe Glu Asn Pro Ala Leu Ala Lys Gly Trp Asp Lys Asn
530 535 540
Lys Glu Gln Asn Asn Thr Ala Ile Ile Leu Met Lys Asp Gly Lys Tyr
545 550 555 560
Tyr Leu Gly Val Ile Asp Lys Asn Asn Lys Leu Arg Ala Asp Asp Leu
565 570 575
Ala Asp Asp Gly Ser Ala Tyr Gly Tyr Met Lys Met Asn Tyr Lys Phe
580 585 590
Ile Pro Thr Pro His Met Glu Leu Pro Lys Val Phe Leu Pro Lys Arg
595 600 605
Ala Pro Lys Arg Tyr Asn Pro Ser Arg Glu Ile Leu Leu Ile Lys Glu
610 615 620
Asn Lys Thr Phe Ile Lys Asp Lys Asn Phe Asn Arg Thr Asp Cys His
625 630 635 640
Lys Leu Ile Asp Phe Phe Lys Asp Ser Ile Asn Lys His Lys Asp Trp
645 650 655
Arg Thr Phe Gly Phe Asp Phe Ser Asp Thr Asp Ser Tyr Glu Asp Ile
660 665 670
Ser Asp Phe Tyr Met Glu Val Gln Asp Gln Gly Tyr Lys Leu Thr Phe
675 680 685
Thr Arg Leu Ser Ala Glu Lys Ile Asp Lys Trp Val Glu Glu Gly Arg
690 695 700
Leu Phe Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ala Asp Gly Ala Gln
705 710 715 720
Gly Ser Pro Asn Leu His Thr Leu Tyr Trp Lys Ala Ile Phe Ser Glu
725 730 735
Glu Asn Leu Lys Asp Val Val Leu Lys Leu Asn Gly Glu Ala Glu Leu
740 745 750
Phe Phe Arg Arg Lys Ser Ile Asp Lys Pro Ala Val His Ala Lys Gly
755 760 765
Ser Met Lys Val Asn Arg Arg Asp Ile Asp Gly Asn Pro Ile Asp Glu
770 775 780
Gly Thr Tyr Val Glu Ile Cys Gly Tyr Ala Asn Gly Lys Arg Asp Met
785 790 795 800
Ala Ser Leu Asn Ala Gly Ala Arg Gly Leu Ile Glu Ser Gly Leu Val
805 810 815
Arg Ile Thr Glu Val Lys His Glu Leu Val Lys Asp Lys Arg Tyr Thr
820 825 830
Ile Asp Lys Tyr Phe Phe His Val Pro Phe Thr Ile Asn Phe Lys Ala
835 840 845
Gln Gly Gln Gly Asn Ile Asn Ser Asp Val Asn Leu Phe Leu Arg Asn
850 855 860
Asn Lys Asp Val Asn Ile Ile Gly Ile Asp Arg Gly Glu Arg Asn Leu
865 870 875 880
Val Tyr Val Ser Leu Ile Asp Arg Asp Gly His Ile Lys Leu Gln Lys
885 890 895
Asp Phe Asn Ile Ile Gly Gly Met Asp Tyr His Ala Lys Leu Asn Gln
900 905 910
Lys Glu Lys Glu Arg Asp Thr Ala Arg Lys Ser Trp Lys Thr Ile Gly
915 920 925
Thr Ile Lys Glu Leu Lys Glu Gly Tyr Leu Ser Gln Val Val His Glu
930 935 940
Ile Val Arg Leu Ala Val Asp Asn Asn Ala Val Ile Val Met Glu Asp
945 950 955 960
Leu Asn Ile Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Val
965 970 975
Tyr Gln Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Leu Val
980 985 990
Phe Lys Asp Ala Gly Tyr Asp Ala Pro Cys Gly Ile Leu Lys Gly Leu
995 1000 1005
Gln Leu Thr Glu Lys Phe Glu Ser Phe Thr Lys Leu Gly Lys Gln
1010 1015 1020
Cys Gly Ile Ile Phe Tyr Ile Pro Ala Gly Tyr Thr Ser Lys Ile
1025 1030 1035
Asp Pro Thr Thr Gly Phe Val Asn Leu Phe Asn Ile Asn Asp Val
1040 1045 1050
Ser Ser Lys Glu Lys Gln Lys Asp Phe Ile Gly Lys Leu Asp Ser
1055 1060 1065
Ile Arg Phe Asp Ala Lys Arg Asp Met Phe Thr Phe Glu Phe Asp
1070 1075 1080
Tyr Asp Lys Phe Arg Thr Tyr Gln Thr Ser Tyr Arg Lys Lys Trp
1085 1090 1095
Ala Val Trp Thr Asn Gly Lys Arg Ile Val Arg Glu Lys Asp Lys
1100 1105 1110
Asp Gly Lys Phe Arg Met Asn Asp Arg Leu Leu Thr Glu Asp Met
1115 1120 1125
Lys Asn Ile Leu Asn Lys Tyr Ala Leu Ala Tyr Lys Ala Gly Glu
1130 1135 1140
Asp Ile Leu Pro Asp Val Ile Ser Arg Asp Lys Ser Leu Ala Ser
1145 1150 1155
Glu Ile Phe Tyr Val Phe Lys Asn Thr Leu Gln Met Arg Asn Ser
1160 1165 1170
Lys Arg Asp Thr Gly Glu Asp Phe Ile Ile Ser Pro Val Leu Asn
1175 1180 1185
Ala Lys Gly Arg Phe Phe Asp Ser Arg Lys Thr Asp Ala Ala Leu
1190 1195 1200
Pro Ile Asp Ala Asp Ala Asn Gly Ala Tyr His Ile Ala Leu Lys
1205 1210 1215
Gly Ser Leu Val Leu Asp Ala Ile Asp Glu Lys Leu Lys Glu Asp
1220 1225 1230
Gly Arg Ile Asp Tyr Lys Asp Met Ala Val Ser Asn Pro Lys Trp
1235 1240 1245
Phe Glu Phe Met Gln Thr Arg Lys Phe Asp Phe
1250 1255
<210> 46
<211> 1263
<212> PRT
<213> 稻田氏钩端螺旋体
<220>
<221> MISC_FEATURE
<222> (1)..(1263)
<223> Genbank WP_020988726 Cpf1
<400> 46
Met Glu Asp Tyr Ser Gly Phe Val Asn Ile Tyr Ser Ile Gln Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Lys Pro Val Gly Lys Thr Leu Glu His Ile Glu
20 25 30
Lys Lys Gly Phe Leu Lys Lys Asp Lys Ile Arg Ala Glu Asp Tyr Lys
35 40 45
Ala Val Lys Lys Ile Ile Asp Lys Tyr His Arg Ala Tyr Ile Glu Glu
50 55 60
Val Phe Asp Ser Val Leu His Gln Lys Lys Lys Lys Asp Lys Thr Arg
65 70 75 80
Phe Ser Thr Gln Phe Ile Lys Glu Ile Lys Glu Phe Ser Glu Leu Tyr
85 90 95
Tyr Lys Thr Glu Lys Asn Ile Pro Asp Lys Glu Arg Leu Glu Ala Leu
100 105 110
Ser Glu Lys Leu Arg Lys Met Leu Val Gly Ala Phe Lys Gly Glu Phe
115 120 125
Ser Glu Glu Val Ala Glu Lys Tyr Lys Asn Leu Phe Ser Lys Glu Leu
130 135 140
Ile Arg Asn Glu Ile Glu Lys Phe Cys Glu Thr Asp Glu Glu Arg Lys
145 150 155 160
Gln Val Ser Asn Phe Lys Ser Phe Thr Thr Tyr Phe Thr Gly Phe His
165 170 175
Ser Asn Arg Gln Asn Ile Tyr Ser Asp Glu Lys Lys Ser Thr Ala Ile
180 185 190
Gly Tyr Arg Ile Ile His Gln Asn Leu Pro Lys Phe Leu Asp Asn Leu
195 200 205
Lys Ile Ile Glu Ser Ile Gln Arg Arg Phe Lys Asp Phe Pro Trp Ser
210 215 220
Asp Leu Lys Lys Asn Leu Lys Lys Ile Asp Lys Asn Ile Lys Leu Thr
225 230 235 240
Glu Tyr Phe Ser Ile Asp Gly Phe Val Asn Val Leu Asn Gln Lys Gly
245 250 255
Ile Asp Ala Tyr Asn Thr Ile Leu Gly Gly Lys Ser Glu Glu Ser Gly
260 265 270
Glu Lys Ile Gln Gly Leu Asn Glu Tyr Ile Asn Leu Tyr Arg Gln Lys
275 280 285
Asn Asn Ile Asp Arg Lys Asn Leu Pro Asn Val Lys Ile Leu Phe Lys
290 295 300
Gln Ile Leu Gly Asp Arg Glu Thr Lys Ser Phe Ile Pro Glu Ala Phe
305 310 315 320
Pro Asp Asp Gln Ser Val Leu Asn Ser Ile Thr Glu Phe Ala Lys Tyr
325 330 335
Leu Lys Leu Asp Lys Lys Lys Lys Ser Ile Ile Ala Glu Leu Lys Lys
340 345 350
Phe Leu Ser Ser Phe Asn Arg Tyr Glu Leu Asp Gly Ile Tyr Leu Ala
355 360 365
Asn Asp Asn Ser Leu Ala Ser Ile Ser Thr Phe Leu Phe Asp Asp Trp
370 375 380
Ser Phe Ile Lys Lys Ser Val Ser Phe Lys Tyr Asp Glu Ser Val Gly
385 390 395 400
Asp Pro Lys Lys Lys Ile Lys Ser Pro Leu Lys Tyr Glu Lys Glu Lys
405 410 415
Glu Lys Trp Leu Lys Gln Lys Tyr Tyr Thr Ile Ser Phe Leu Asn Asp
420 425 430
Ala Ile Glu Ser Tyr Ser Lys Ser Gln Asp Glu Lys Arg Val Lys Ile
435 440 445
Arg Leu Glu Ala Tyr Phe Ala Glu Phe Lys Ser Lys Asp Asp Ala Lys
450 455 460
Lys Gln Phe Asp Leu Leu Glu Arg Ile Glu Glu Ala Tyr Ala Ile Val
465 470 475 480
Glu Pro Leu Leu Gly Ala Glu Tyr Pro Arg Asp Arg Asn Leu Lys Ala
485 490 495
Asp Lys Lys Glu Val Gly Lys Ile Lys Asp Phe Leu Asp Ser Ile Lys
500 505 510
Ser Leu Gln Phe Phe Leu Lys Pro Leu Leu Ser Ala Glu Ile Phe Asp
515 520 525
Glu Lys Asp Leu Gly Phe Tyr Asn Gln Leu Glu Gly Tyr Tyr Glu Glu
530 535 540
Ile Asp Ser Ile Gly His Leu Tyr Asn Lys Val Arg Asn Tyr Leu Thr
545 550 555 560
Gly Lys Ile Tyr Ser Lys Glu Lys Phe Lys Leu Asn Phe Glu Asn Ser
565 570 575
Thr Leu Leu Lys Gly Trp Asp Glu Asn Arg Glu Val Ala Asn Leu Cys
580 585 590
Val Ile Phe Arg Glu Asp Gln Lys Tyr Tyr Leu Gly Val Met Asp Lys
595 600 605
Glu Asn Asn Thr Ile Leu Ser Asp Ile Pro Lys Val Lys Pro Asn Glu
610 615 620
Leu Phe Tyr Glu Lys Met Val Tyr Lys Leu Ile Pro Thr Pro His Met
625 630 635 640
Gln Leu Pro Arg Ile Ile Phe Ser Ser Asp Asn Leu Ser Ile Tyr Asn
645 650 655
Pro Ser Lys Ser Ile Leu Lys Ile Arg Glu Ala Lys Ser Phe Lys Glu
660 665 670
Gly Lys Asn Phe Lys Leu Lys Asp Cys His Lys Phe Ile Asp Phe Tyr
675 680 685
Lys Glu Ser Ile Ser Lys Asn Glu Asp Trp Ser Arg Phe Asp Phe Lys
690 695 700
Phe Ser Lys Thr Ser Ser Tyr Glu Asn Ile Ser Glu Phe Tyr Arg Glu
705 710 715 720
Val Glu Arg Gln Gly Tyr Asn Leu Asp Phe Lys Lys Val Ser Lys Phe
725 730 735
Tyr Ile Asp Ser Leu Val Glu Asp Gly Lys Leu Tyr Leu Phe Gln Ile
740 745 750
Tyr Asn Lys Asp Phe Ser Ile Phe Ser Lys Gly Lys Pro Asn Leu His
755 760 765
Thr Ile Tyr Phe Arg Ser Leu Phe Ser Lys Glu Asn Leu Lys Asp Val
770 775 780
Cys Leu Lys Leu Asn Gly Glu Ala Glu Met Phe Phe Arg Lys Lys Ser
785 790 795 800
Ile Asn Tyr Asp Glu Lys Lys Lys Arg Glu Gly His His Pro Glu Leu
805 810 815
Phe Glu Lys Leu Lys Tyr Pro Ile Leu Lys Asp Lys Arg Tyr Ser Glu
820 825 830
Asp Lys Phe Gln Phe His Leu Pro Ile Ser Leu Asn Phe Lys Ser Lys
835 840 845
Glu Arg Leu Asn Phe Asn Leu Lys Val Asn Glu Phe Leu Lys Arg Asn
850 855 860
Lys Asp Ile Asn Ile Ile Gly Ile Asp Arg Gly Glu Arg Asn Leu Leu
865 870 875 880
Tyr Leu Val Met Ile Asn Gln Lys Gly Glu Ile Leu Lys Gln Thr Leu
885 890 895
Leu Asp Ser Met Gln Ser Gly Lys Gly Arg Pro Glu Ile Asn Tyr Lys
900 905 910
Glu Lys Leu Gln Glu Lys Glu Ile Glu Arg Asp Lys Ala Arg Lys Ser
915 920 925
Trp Gly Thr Val Glu Asn Ile Lys Glu Leu Lys Glu Gly Tyr Leu Ser
930 935 940
Ile Val Ile His Gln Ile Ser Lys Leu Met Val Glu Asn Asn Ala Ile
945 950 955 960
Val Val Leu Glu Asp Leu Asn Ile Gly Phe Lys Arg Gly Arg Gln Lys
965 970 975
Val Glu Arg Gln Val Tyr Gln Lys Phe Glu Lys Met Leu Ile Asp Lys
980 985 990
Leu Asn Phe Leu Val Phe Lys Glu Asn Lys Pro Thr Glu Pro Gly Gly
995 1000 1005
Val Leu Lys Ala Tyr Gln Leu Thr Asp Glu Phe Gln Ser Phe Glu
1010 1015 1020
Lys Leu Ser Lys Gln Thr Gly Phe Leu Phe Tyr Val Pro Ser Trp
1025 1030 1035
Asn Thr Ser Lys Ile Asp Pro Arg Thr Gly Phe Ile Asp Phe Leu
1040 1045 1050
His Pro Ala Tyr Glu Asn Ile Glu Lys Ala Lys Gln Trp Ile Asn
1055 1060 1065
Lys Phe Asp Ser Ile Arg Phe Asn Ser Lys Met Asp Trp Phe Glu
1070 1075 1080
Phe Thr Ala Asp Thr Arg Lys Phe Ser Glu Asn Leu Met Leu Gly
1085 1090 1095
Lys Asn Arg Val Trp Val Ile Cys Thr Thr Asn Val Glu Arg Tyr
1100 1105 1110
Phe Thr Ser Lys Thr Ala Asn Ser Ser Ile Gln Tyr Asn Ser Ile
1115 1120 1125
Gln Ile Thr Glu Lys Leu Lys Glu Leu Phe Val Asp Ile Pro Phe
1130 1135 1140
Ser Asn Gly Gln Asp Leu Lys Pro Glu Ile Leu Arg Lys Asn Asp
1145 1150 1155
Ala Val Phe Phe Lys Ser Leu Leu Phe Tyr Ile Lys Thr Thr Leu
1160 1165 1170
Ser Leu Arg Gln Asn Asn Gly Lys Lys Gly Glu Glu Glu Lys Asp
1175 1180 1185
Phe Ile Leu Ser Pro Val Val Asp Ser Lys Gly Arg Phe Phe Asn
1190 1195 1200
Ser Leu Glu Ala Ser Asp Asp Glu Pro Lys Asp Ala Asp Ala Asn
1205 1210 1215
Gly Ala Tyr His Ile Ala Leu Lys Gly Leu Met Asn Leu Leu Val
1220 1225 1230
Leu Asn Glu Thr Lys Glu Glu Asn Leu Ser Arg Pro Lys Trp Lys
1235 1240 1245
Ile Lys Asn Lys Asp Trp Leu Glu Phe Val Trp Glu Arg Asn Arg
1250 1255 1260
<210> 47
<211> 1260
<212> PRT
<213> 狗齿龈卟啉单胞菌
<220>
<221> MISC_FEATURE
<222> (1)..(1260)
<223> Genbank WP_023941260 Cpf1
<400> 47
Met Asp Ser Leu Lys Asp Phe Thr Asn Leu Tyr Pro Val Ser Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Lys Pro Val Gly Lys Thr Leu Glu Asn Ile Glu
20 25 30
Lys Ala Gly Ile Leu Lys Glu Asp Glu His Arg Ala Glu Ser Tyr Arg
35 40 45
Arg Val Lys Lys Ile Ile Asp Thr Tyr His Lys Val Phe Ile Asp Ser
50 55 60
Ser Leu Glu Asn Met Ala Lys Met Gly Ile Glu Asn Glu Ile Lys Ala
65 70 75 80
Met Leu Gln Ser Phe Cys Glu Leu Tyr Lys Lys Asp His Arg Thr Glu
85 90 95
Gly Glu Asp Lys Ala Leu Asp Lys Ile Arg Ala Val Leu Arg Gly Leu
100 105 110
Ile Val Gly Ala Phe Thr Gly Val Cys Gly Arg Arg Glu Asn Thr Val
115 120 125
Gln Asn Glu Lys Tyr Glu Ser Leu Phe Lys Glu Lys Leu Ile Lys Glu
130 135 140
Ile Leu Pro Asp Phe Val Leu Ser Thr Glu Ala Glu Ser Leu Pro Phe
145 150 155 160
Ser Val Glu Glu Ala Thr Arg Ser Leu Lys Glu Phe Asp Ser Phe Thr
165 170 175
Ser Tyr Phe Ala Gly Phe Tyr Glu Asn Arg Lys Asn Ile Tyr Ser Thr
180 185 190
Lys Pro Gln Ser Thr Ala Ile Ala Tyr Arg Leu Ile His Glu Asn Leu
195 200 205
Pro Lys Phe Ile Asp Asn Ile Leu Val Phe Gln Lys Ile Lys Glu Pro
210 215 220
Ile Ala Lys Glu Leu Glu His Ile Arg Ala Asp Phe Ser Ala Gly Gly
225 230 235 240
Tyr Ile Lys Lys Asp Glu Arg Leu Glu Asp Ile Phe Ser Leu Asn Tyr
245 250 255
Tyr Ile His Val Leu Ser Gln Ala Gly Ile Glu Lys Tyr Asn Ala Leu
260 265 270
Ile Gly Lys Ile Val Thr Glu Gly Asp Gly Glu Met Lys Gly Leu Asn
275 280 285
Glu His Ile Asn Leu Tyr Asn Gln Gln Arg Gly Arg Glu Asp Arg Leu
290 295 300
Pro Leu Phe Arg Pro Leu Tyr Lys Gln Ile Leu Ser Asp Arg Glu Gln
305 310 315 320
Leu Ser Tyr Leu Pro Glu Ser Phe Glu Lys Asp Glu Glu Leu Leu Arg
325 330 335
Ala Leu Lys Glu Phe Tyr Asp His Ile Ala Glu Asp Ile Leu Gly Arg
340 345 350
Thr Gln Gln Leu Met Thr Ser Ile Ser Glu Tyr Asp Leu Ser Arg Ile
355 360 365
Tyr Val Arg Asn Asp Ser Gln Leu Thr Asp Ile Ser Lys Lys Met Leu
370 375 380
Gly Asp Trp Asn Ala Ile Tyr Met Ala Arg Glu Arg Ala Tyr Asp His
385 390 395 400
Glu Gln Ala Pro Lys Arg Ile Thr Ala Lys Tyr Glu Arg Asp Arg Ile
405 410 415
Lys Ala Leu Lys Gly Glu Glu Ser Ile Ser Leu Ala Asn Leu Asn Ser
420 425 430
Cys Ile Ala Phe Leu Asp Asn Val Arg Asp Cys Arg Val Asp Thr Tyr
435 440 445
Leu Ser Thr Leu Gly Gln Lys Glu Gly Pro His Gly Leu Ser Asn Leu
450 455 460
Val Glu Asn Val Phe Ala Ser Tyr His Glu Ala Glu Gln Leu Leu Ser
465 470 475 480
Phe Pro Tyr Pro Glu Glu Asn Asn Leu Ile Gln Asp Lys Asp Asn Val
485 490 495
Val Leu Ile Lys Asn Leu Leu Asp Asn Ile Ser Asp Leu Gln Arg Phe
500 505 510
Leu Lys Pro Leu Trp Gly Met Gly Asp Glu Pro Asp Lys Asp Glu Arg
515 520 525
Phe Tyr Gly Glu Tyr Asn Tyr Ile Arg Gly Ala Leu Asp Gln Val Ile
530 535 540
Pro Leu Tyr Asn Lys Val Arg Asn Tyr Leu Thr Arg Lys Pro Tyr Ser
545 550 555 560
Thr Arg Lys Val Lys Leu Asn Phe Gly Asn Ser Gln Leu Leu Ser Gly
565 570 575
Trp Asp Arg Asn Lys Glu Lys Asp Asn Ser Cys Val Ile Leu Arg Lys
580 585 590
Gly Gln Asn Phe Tyr Leu Ala Ile Met Asn Asn Arg His Lys Arg Ser
595 600 605
Phe Glu Asn Lys Val Leu Pro Glu Tyr Lys Glu Gly Glu Pro Tyr Phe
610 615 620
Glu Lys Met Asp Tyr Lys Phe Leu Pro Asp Pro Asn Lys Met Leu Pro
625 630 635 640
Lys Val Phe Leu Ser Lys Lys Gly Ile Glu Ile Tyr Lys Pro Ser Pro
645 650 655
Lys Leu Leu Glu Gln Tyr Gly His Gly Thr His Lys Lys Gly Asp Thr
660 665 670
Phe Ser Met Asp Asp Leu His Glu Leu Ile Asp Phe Phe Lys His Ser
675 680 685
Ile Glu Ala His Glu Asp Trp Lys Gln Phe Gly Phe Lys Phe Ser Asp
690 695 700
Thr Ala Thr Tyr Glu Asn Val Ser Ser Phe Tyr Arg Glu Val Glu Asp
705 710 715 720
Gln Gly Tyr Lys Leu Ser Phe Arg Lys Val Ser Glu Ser Tyr Val Tyr
725 730 735
Ser Leu Ile Asp Gln Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys
740 745 750
Asp Phe Ser Pro Cys Ser Lys Gly Thr Pro Asn Leu His Thr Leu Tyr
755 760 765
Trp Arg Met Leu Phe Asp Glu Arg Asn Leu Ala Asp Val Ile Tyr Lys
770 775 780
Leu Asp Gly Lys Ala Glu Ile Phe Phe Arg Glu Lys Ser Leu Lys Asn
785 790 795 800
Asp His Pro Thr His Pro Ala Gly Lys Pro Ile Lys Lys Lys Ser Arg
805 810 815
Gln Lys Lys Gly Glu Glu Ser Leu Phe Glu Tyr Asp Leu Val Lys Asp
820 825 830
Arg Arg Tyr Thr Met Asp Lys Phe Gln Phe His Val Pro Ile Thr Met
835 840 845
Asn Phe Lys Cys Ser Ala Gly Ser Lys Val Asn Asp Met Val Asn Ala
850 855 860
His Ile Arg Glu Ala Lys Asp Met His Val Ile Gly Ile Asp Arg Gly
865 870 875 880
Glu Arg Asn Leu Leu Tyr Ile Cys Val Ile Asp Ser Arg Gly Thr Ile
885 890 895
Leu Asp Gln Ile Ser Leu Asn Thr Ile Asn Asp Ile Asp Tyr His Asp
900 905 910
Leu Leu Glu Ser Arg Asp Lys Asp Arg Gln Gln Glu Arg Arg Asn Trp
915 920 925
Gln Thr Ile Glu Gly Ile Lys Glu Leu Lys Gln Gly Tyr Leu Ser Gln
930 935 940
Ala Val His Arg Ile Ala Glu Leu Met Val Ala Tyr Lys Ala Val Val
945 950 955 960
Ala Leu Glu Asp Leu Asn Met Gly Phe Lys Arg Gly Arg Gln Lys Val
965 970 975
Glu Ser Ser Val Tyr Gln Gln Phe Glu Lys Gln Leu Ile Asp Lys Leu
980 985 990
Asn Tyr Leu Val Asp Lys Lys Lys Arg Pro Glu Asp Ile Gly Gly Leu
995 1000 1005
Leu Arg Ala Tyr Gln Phe Thr Ala Pro Phe Lys Ser Phe Lys Glu
1010 1015 1020
Met Gly Lys Gln Asn Gly Phe Leu Phe Tyr Ile Pro Ala Trp Asn
1025 1030 1035
Thr Ser Asn Ile Asp Pro Thr Thr Gly Phe Val Asn Leu Phe His
1040 1045 1050
Ala Gln Tyr Glu Asn Val Asp Lys Ala Lys Ser Phe Phe Gln Lys
1055 1060 1065
Phe Asp Ser Ile Ser Tyr Asn Pro Lys Lys Asp Trp Phe Glu Phe
1070 1075 1080
Ala Phe Asp Tyr Lys Asn Phe Thr Lys Lys Ala Glu Gly Ser Arg
1085 1090 1095
Ser Met Trp Ile Leu Cys Thr His Gly Ser Arg Ile Lys Asn Phe
1100 1105 1110
Arg Asn Ser Gln Lys Asn Gly Gln Trp Asp Ser Glu Glu Phe Ala
1115 1120 1125
Leu Thr Glu Ala Phe Lys Ser Leu Phe Val Arg Tyr Glu Ile Asp
1130 1135 1140
Tyr Thr Ala Asp Leu Lys Thr Ala Ile Val Asp Glu Lys Gln Lys
1145 1150 1155
Asp Phe Phe Val Asp Leu Leu Lys Leu Phe Lys Leu Thr Val Gln
1160 1165 1170
Met Arg Asn Ser Trp Lys Glu Lys Asp Leu Asp Tyr Leu Ile Ser
1175 1180 1185
Pro Val Ala Gly Ala Asp Gly Arg Phe Phe Asp Thr Arg Glu Gly
1190 1195 1200
Asn Lys Ser Leu Pro Lys Asp Ala Asp Ala Asn Gly Ala Tyr Asn
1205 1210 1215
Ile Ala Leu Lys Gly Leu Trp Ala Leu Arg Gln Ile Arg Gln Thr
1220 1225 1230
Ser Glu Gly Gly Lys Leu Lys Leu Ala Ile Ser Asn Lys Glu Trp
1235 1240 1245
Leu Gln Phe Val Gln Glu Arg Ser Tyr Glu Lys Asp
1250 1255 1260
<210> 48
<211> 1253
<212> PRT
<213> 阿尔巴普雷沃氏菌
<220>
<221> MISC_FEATURE
<222> (1)..(1253)
<223> Genbank WP_024988992 Cpf1
<400> 48
Met Asn Ile Lys Asn Phe Thr Gly Leu Tyr Pro Leu Ser Lys Thr Leu
1 5 10 15
Arg Phe Glu Leu Lys Pro Ile Gly Lys Thr Lys Glu Asn Ile Glu Lys
20 25 30
Asn Gly Ile Leu Thr Lys Asp Glu Gln Arg Ala Lys Asp Tyr Leu Ile
35 40 45
Val Lys Gly Phe Ile Asp Glu Tyr His Lys Gln Phe Ile Lys Asp Arg
50 55 60
Leu Trp Asp Phe Lys Leu Pro Leu Glu Ser Glu Gly Glu Lys Asn Ser
65 70 75 80
Leu Glu Glu Tyr Gln Glu Leu Tyr Glu Leu Thr Lys Arg Asn Asp Ala
85 90 95
Gln Glu Ala Asp Phe Thr Glu Ile Lys Asp Asn Leu Arg Ser Ser Ile
100 105 110
Thr Glu Gln Leu Thr Lys Ser Gly Ser Ala Tyr Asp Arg Ile Phe Lys
115 120 125
Lys Glu Phe Ile Arg Glu Asp Leu Val Asn Phe Leu Glu Asp Glu Lys
130 135 140
Asp Lys Asn Ile Val Lys Gln Phe Glu Asp Phe Thr Thr Tyr Phe Thr
145 150 155 160
Gly Phe Tyr Glu Asn Arg Lys Asn Met Tyr Ser Ser Glu Glu Lys Ser
165 170 175
Thr Ala Ile Ala Tyr Arg Leu Ile His Gln Asn Leu Pro Lys Phe Met
180 185 190
Asp Asn Met Arg Ser Phe Ala Lys Ile Ala Asn Ser Ser Val Ser Glu
195 200 205
His Phe Ser Asp Ile Tyr Glu Ser Trp Lys Glu Tyr Leu Asn Val Asn
210 215 220
Ser Ile Glu Glu Ile Phe Gln Leu Asp Tyr Phe Ser Glu Thr Leu Thr
225 230 235 240
Gln Pro His Ile Glu Val Tyr Asn Tyr Ile Ile Gly Lys Lys Val Leu
245 250 255
Glu Asp Gly Thr Glu Ile Lys Gly Ile Asn Glu Tyr Val Asn Leu Tyr
260 265 270
Asn Gln Gln Gln Lys Asp Lys Ser Lys Arg Leu Pro Phe Leu Val Pro
275 280 285
Leu Tyr Lys Gln Ile Leu Ser Asp Arg Glu Lys Leu Ser Trp Ile Ala
290 295 300
Glu Glu Phe Asp Ser Asp Lys Lys Met Leu Ser Ala Ile Thr Glu Ser
305 310 315 320
Tyr Asn His Leu His Asn Val Leu Met Gly Asn Glu Asn Glu Ser Leu
325 330 335
Arg Asn Leu Leu Leu Asn Ile Lys Asp Tyr Asn Leu Glu Lys Ile Asn
340 345 350
Ile Thr Asn Asp Leu Ser Leu Thr Glu Ile Ser Gln Asn Leu Phe Gly
355 360 365
Arg Tyr Asp Val Phe Thr Asn Gly Ile Lys Asn Lys Leu Arg Val Leu
370 375 380
Thr Pro Arg Lys Lys Lys Glu Thr Asp Glu Asn Phe Glu Asp Arg Ile
385 390 395 400
Asn Lys Ile Phe Lys Thr Gln Lys Ser Phe Ser Ile Ala Phe Leu Asn
405 410 415
Lys Leu Pro Gln Pro Glu Met Glu Asp Gly Lys Pro Arg Asn Ile Glu
420 425 430
Asp Tyr Phe Ile Thr Gln Gly Ala Ile Asn Thr Lys Ser Ile Gln Lys
435 440 445
Glu Asp Ile Phe Ala Gln Ile Glu Asn Ala Tyr Glu Asp Ala Gln Val
450 455 460
Phe Leu Gln Ile Lys Asp Thr Asp Asn Lys Leu Ser Gln Asn Lys Thr
465 470 475 480
Ala Val Glu Lys Ile Lys Thr Leu Leu Asp Ala Leu Lys Glu Leu Gln
485 490 495
His Phe Ile Lys Pro Leu Leu Gly Ser Gly Glu Glu Asn Glu Lys Asp
500 505 510
Glu Leu Phe Tyr Gly Ser Phe Leu Ala Ile Trp Asp Glu Leu Asp Thr
515 520 525
Ile Thr Pro Leu Tyr Asn Lys Val Arg Asn Trp Leu Thr Arg Lys Pro
530 535 540
Tyr Ser Thr Glu Lys Ile Lys Leu Asn Phe Asp Asn Ala Gln Leu Leu
545 550 555 560
Gly Gly Trp Asp Val Asn Lys Glu His Asp Cys Ala Gly Ile Leu Leu
565 570 575
Arg Lys Asn Asp Ser Tyr Tyr Leu Gly Ile Ile Asn Lys Lys Thr Asn
580 585 590
His Ile Phe Asp Thr Asp Ile Thr Pro Ser Asp Gly Glu Cys Tyr Asp
595 600 605
Lys Ile Asp Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro Lys
610 615 620
Val Phe Phe Ser Lys Ser Arg Ile Lys Glu Phe Glu Pro Ser Glu Ala
625 630 635 640
Ile Ile Asn Cys Tyr Lys Lys Gly Thr His Lys Lys Gly Lys Asn Phe
645 650 655
Asn Leu Thr Asp Cys His Arg Leu Ile Asn Phe Phe Lys Thr Ser Ile
660 665 670
Glu Lys His Glu Asp Trp Ser Lys Phe Gly Phe Lys Phe Ser Asp Thr
675 680 685
Glu Thr Tyr Glu Asp Ile Ser Gly Phe Tyr Arg Glu Val Glu Gln Gln
690 695 700
Gly Tyr Arg Leu Thr Ser His Pro Val Ser Ala Ser Tyr Ile His Ser
705 710 715 720
Leu Val Lys Glu Gly Lys Leu Tyr Leu Phe Gln Ile Trp Asn Lys Asp
725 730 735
Phe Ser Gln Phe Ser Lys Gly Thr Pro Asn Leu His Thr Leu Tyr Trp
740 745 750
Lys Met Leu Phe Asp Lys Arg Asn Leu Ser Asp Val Val Tyr Lys Leu
755 760 765
Asn Gly Gln Ala Glu Val Phe Tyr Arg Lys Ser Ser Ile Glu His Gln
770 775 780
Asn Arg Ile Ile His Pro Ala Gln His Pro Ile Thr Asn Lys Asn Glu
785 790 795 800
Leu Asn Lys Lys His Thr Ser Thr Phe Lys Tyr Asp Ile Ile Lys Asp
805 810 815
Arg Arg Tyr Thr Val Asp Lys Phe Gln Phe His Val Pro Ile Thr Ile
820 825 830
Asn Phe Lys Ala Thr Gly Gln Asn Asn Ile Asn Pro Ile Val Gln Glu
835 840 845
Val Ile Arg Gln Asn Gly Ile Thr His Ile Ile Gly Ile Asp Arg Gly
850 855 860
Glu Arg His Leu Leu Tyr Leu Ser Leu Ile Asp Leu Lys Gly Asn Ile
865 870 875 880
Ile Lys Gln Met Thr Leu Asn Glu Ile Ile Asn Glu Tyr Lys Gly Val
885 890 895
Thr Tyr Lys Thr Asn Tyr His Asn Leu Leu Glu Lys Arg Glu Lys Glu
900 905 910
Arg Thr Glu Ala Arg His Ser Trp Ser Ser Ile Glu Ser Ile Lys Glu
915 920 925
Leu Lys Asp Gly Tyr Met Ser Gln Val Ile His Lys Ile Thr Asp Met
930 935 940
Met Val Lys Tyr Asn Ala Ile Val Val Leu Glu Asp Leu Asn Gly Gly
945 950 955 960
Phe Met Arg Gly Arg Gln Lys Val Glu Lys Gln Val Tyr Gln Lys Phe
965 970 975
Glu Lys Lys Leu Ile Asp Lys Leu Asn Tyr Leu Val Asp Lys Lys Leu
980 985 990
Asp Ala Asn Glu Val Gly Gly Val Leu Asn Ala Tyr Gln Leu Thr Asn
995 1000 1005
Lys Phe Glu Ser Phe Lys Lys Ile Gly Lys Gln Ser Gly Phe Leu
1010 1015 1020
Phe Tyr Ile Pro Ala Trp Asn Thr Ser Lys Ile Asp Pro Ile Thr
1025 1030 1035
Gly Phe Val Asn Leu Phe Asn Thr Arg Tyr Glu Ser Ile Lys Glu
1040 1045 1050
Thr Lys Val Phe Trp Ser Lys Phe Asp Ile Ile Arg Tyr Asn Lys
1055 1060 1065
Glu Lys Asn Trp Phe Glu Phe Val Phe Asp Tyr Asn Thr Phe Thr
1070 1075 1080
Thr Lys Ala Glu Gly Thr Arg Thr Lys Trp Thr Leu Cys Thr His
1085 1090 1095
Gly Thr Arg Ile Gln Thr Phe Arg Asn Pro Glu Lys Asn Ala Gln
1100 1105 1110
Trp Asp Asn Lys Glu Ile Asn Leu Thr Glu Ser Phe Lys Ala Leu
1115 1120 1125
Phe Glu Lys Tyr Lys Ile Asp Ile Thr Ser Asn Leu Lys Glu Ser
1130 1135 1140
Ile Met Gln Glu Thr Glu Lys Lys Phe Phe Gln Glu Leu His Asn
1145 1150 1155
Leu Leu His Leu Thr Leu Gln Met Arg Asn Ser Val Thr Gly Thr
1160 1165 1170
Asp Ile Asp Tyr Leu Ile Ser Pro Val Ala Asp Glu Asp Gly Asn
1175 1180 1185
Phe Tyr Asp Ser Arg Ile Asn Gly Lys Asn Phe Pro Glu Asn Ala
1190 1195 1200
Asp Ala Asn Gly Ala Tyr Asn Ile Ala Arg Lys Gly Leu Met Leu
1205 1210 1215
Ile Arg Gln Ile Lys Gln Ala Asp Pro Gln Lys Lys Phe Lys Phe
1220 1225 1230
Glu Thr Ile Thr Asn Lys Asp Trp Leu Lys Phe Ala Gln Asp Lys
1235 1240 1245
Pro Tyr Leu Lys Asp
1250
<210> 49
<211> 1231
<212> PRT
<213> 溶纤维丁酸弧菌
<220>
<221> MISC_FEATURE
<222> (1)..(1231)
<223> Genbank WP_027216152 Cpf1
<400> 49
Met Tyr Tyr Glu Ser Leu Thr Lys Leu Tyr Pro Ile Lys Lys Thr Ile
1 5 10 15
Arg Asn Glu Leu Val Pro Ile Gly Lys Thr Leu Glu Asn Ile Lys Lys
20 25 30
Asn Asn Ile Leu Glu Ala Asp Glu Asp Arg Lys Ile Ala Tyr Ile Arg
35 40 45
Val Lys Ala Ile Met Asp Asp Tyr His Lys Arg Leu Ile Asn Glu Ala
50 55 60
Leu Ser Gly Phe Ala Leu Ile Asp Leu Asp Lys Ala Ala Asn Leu Tyr
65 70 75 80
Leu Ser Arg Ser Lys Ser Ala Asp Asp Ile Glu Ser Phe Ser Arg Phe
85 90 95
Gln Asp Lys Leu Arg Lys Ala Ile Ala Lys Arg Leu Arg Glu His Glu
100 105 110
Asn Phe Gly Lys Ile Gly Asn Lys Asp Ile Ile Pro Leu Leu Gln Lys
115 120 125
Leu Ser Glu Asn Glu Asp Asp Tyr Asn Ala Leu Glu Ser Phe Lys Asn
130 135 140
Phe Tyr Thr Tyr Phe Glu Ser Tyr Asn Asp Val Arg Leu Asn Leu Tyr
145 150 155 160
Ser Asp Lys Glu Lys Ser Ser Thr Val Ala Tyr Arg Leu Ile Asn Glu
165 170 175
Asn Leu Pro Arg Phe Leu Asp Asn Ile Arg Ala Tyr Asp Ala Val Gln
180 185 190
Lys Ala Gly Ile Thr Ser Glu Glu Leu Ser Ser Glu Ala Gln Asp Gly
195 200 205
Leu Phe Leu Val Asn Thr Phe Asn Asn Val Leu Ile Gln Asp Gly Ile
210 215 220
Asn Thr Tyr Asn Glu Asp Ile Gly Lys Leu Asn Val Ala Ile Asn Leu
225 230 235 240
Tyr Asn Gln Lys Asn Ala Ser Val Gln Gly Phe Arg Lys Val Pro Lys
245 250 255
Met Lys Val Leu Tyr Lys Gln Ile Leu Ser Asp Arg Glu Glu Ser Phe
260 265 270
Ile Asp Glu Phe Glu Ser Asp Thr Glu Leu Leu Asp Ser Leu Glu Ser
275 280 285
His Tyr Ala Asn Leu Ala Lys Tyr Phe Gly Ser Asn Lys Val Gln Leu
290 295 300
Leu Phe Thr Ala Leu Arg Glu Ser Lys Gly Val Asn Val Tyr Val Lys
305 310 315 320
Asn Asp Ile Ala Lys Thr Ser Phe Ser Asn Val Val Phe Gly Ser Trp
325 330 335
Ser Arg Ile Asp Glu Leu Ile Asn Gly Glu Tyr Asp Asp Asn Asn Asn
340 345 350
Arg Lys Lys Asp Glu Lys Tyr Tyr Asp Lys Arg Gln Lys Glu Leu Lys
355 360 365
Lys Asn Lys Ser Tyr Thr Ile Glu Lys Ile Ile Thr Leu Ser Thr Glu
370 375 380
Asp Val Asp Val Ile Gly Lys Tyr Ile Glu Lys Leu Glu Ser Asp Ile
385 390 395 400
Asp Asp Ile Arg Phe Lys Gly Lys Asn Phe Tyr Glu Ala Val Leu Cys
405 410 415
Gly His Asp Arg Ser Lys Lys Leu Ser Lys Asn Lys Gly Ala Val Glu
420 425 430
Ala Ile Lys Gly Tyr Leu Asp Ser Val Lys Asp Phe Glu Arg Asp Leu
435 440 445
Lys Leu Ile Asn Gly Ser Gly Gln Glu Leu Glu Lys Asn Leu Val Val
450 455 460
Tyr Gly Glu Gln Glu Ala Val Leu Ser Glu Leu Ser Gly Ile Asp Ser
465 470 475 480
Leu Tyr Asn Met Thr Arg Asn Tyr Leu Thr Lys Lys Pro Phe Ser Thr
485 490 495
Glu Lys Ile Lys Leu Asn Phe Asn Lys Pro Thr Phe Leu Asp Gly Trp
500 505 510
Asp Tyr Gly Asn Glu Glu Ala Tyr Leu Gly Phe Phe Met Ile Lys Glu
515 520 525
Gly Asn Tyr Phe Leu Ala Val Met Asp Ala Asn Trp Asn Lys Glu Phe
530 535 540
Arg Asn Ile Pro Ser Val Asp Lys Ser Asp Cys Tyr Lys Lys Val Ile
545 550 555 560
Tyr Lys Gln Ile Ser Ser Pro Glu Lys Ser Ile Gln Asn Leu Met Val
565 570 575
Ile Asp Gly Lys Thr Val Lys Lys Asn Gly Arg Lys Glu Lys Glu Gly
580 585 590
Ile His Ser Gly Glu Asn Leu Ile Leu Glu Glu Leu Lys Asn Thr Tyr
595 600 605
Leu Pro Lys Lys Ile Asn Asp Ile Arg Lys Arg Arg Ser Tyr Leu Asn
610 615 620
Gly Asp Thr Phe Ser Lys Lys Asp Leu Thr Glu Phe Ile Gly Tyr Tyr
625 630 635 640
Lys Gln Arg Val Ile Glu Tyr Tyr Asn Gly Tyr Ser Phe Tyr Phe Lys
645 650 655
Ser Asp Asp Asp Tyr Ala Ser Phe Lys Glu Phe Gln Glu Asp Val Gly
660 665 670
Arg Gln Ala Tyr Gln Ile Ser Tyr Val Asp Val Pro Val Ser Phe Val
675 680 685
Asp Asp Leu Ile Asn Ser Gly Lys Leu Tyr Leu Phe Arg Val Tyr Asn
690 695 700
Lys Asp Phe Ser Glu Tyr Ser Lys Gly Arg Leu Asn Leu His Thr Leu
705 710 715 720
Tyr Phe Lys Met Leu Phe Asp Glu Arg Asn Leu Lys Asn Val Val Tyr
725 730 735
Lys Leu Asn Gly Gln Ala Glu Val Phe Tyr Arg Pro Ser Ser Ile Lys
740 745 750
Lys Glu Glu Leu Ile Val His Arg Ala Gly Glu Glu Ile Lys Asn Lys
755 760 765
Asn Pro Lys Arg Ala Ala Gln Lys Pro Thr Arg Arg Leu Asp Tyr Asp
770 775 780
Ile Val Lys Asp Arg Arg Tyr Ser Gln Asp Lys Phe Met Leu His Thr
785 790 795 800
Ser Ile Ile Met Asn Phe Gly Ala Glu Glu Asn Val Ser Phe Asn Asp
805 810 815
Ile Val Asn Gly Val Leu Arg Asn Glu Asp Lys Val Asn Val Ile Gly
820 825 830
Ile Asp Arg Gly Glu Arg Asn Leu Leu Tyr Val Val Val Ile Asp Pro
835 840 845
Glu Gly Lys Ile Leu Glu Gln Arg Ser Leu Asn Cys Ile Thr Asp Ser
850 855 860
Asn Leu Asp Ile Glu Thr Asp Tyr His Arg Leu Leu Asp Glu Lys Glu
865 870 875 880
Ser Asp Arg Lys Ile Ala Arg Arg Asp Trp Thr Thr Ile Glu Asn Ile
885 890 895
Lys Glu Leu Lys Ala Gly Tyr Leu Ser Gln Val Val His Ile Val Ala
900 905 910
Glu Leu Val Leu Lys Tyr Asn Ala Ile Ile Cys Leu Glu Asp Leu Asn
915 920 925
Phe Gly Phe Lys Arg Gly Arg Gln Lys Val Glu Lys Gln Val Tyr Gln
930 935 940
Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Leu Val Met Asp
945 950 955 960
Lys Ser Arg Glu Gln Leu Ser Pro Glu Lys Ile Ser Gly Ala Leu Asn
965 970 975
Ala Leu Gln Leu Thr Pro Asp Phe Lys Ser Phe Lys Val Leu Gly Lys
980 985 990
Gln Thr Gly Ile Ile Tyr Tyr Val Pro Ala Tyr Leu Thr Ser Lys Ile
995 1000 1005
Asp Pro Met Thr Gly Phe Ala Asn Leu Phe Tyr Val Lys Tyr Glu
1010 1015 1020
Asn Val Asp Lys Ala Lys Glu Phe Phe Ser Lys Phe Asp Ser Ile
1025 1030 1035
Lys Tyr Asn Lys Asp Gly Lys Asn Trp Asn Thr Lys Gly Tyr Phe
1040 1045 1050
Glu Phe Ala Phe Asp Tyr Lys Lys Phe Thr Asp Arg Ala Tyr Gly
1055 1060 1065
Arg Val Ser Glu Trp Thr Val Cys Thr Val Gly Glu Arg Ile Ile
1070 1075 1080
Lys Phe Lys Asn Lys Glu Lys Asn Asn Ser Tyr Asp Asp Lys Val
1085 1090 1095
Ile Asp Leu Thr Asn Ser Leu Lys Glu Leu Phe Asp Ser Tyr Lys
1100 1105 1110
Val Thr Tyr Glu Ser Glu Val Asp Leu Lys Asp Ala Ile Leu Ala
1115 1120 1125
Ile Asp Asp Pro Ala Phe Tyr Arg Asp Leu Thr Arg Arg Leu Gln
1130 1135 1140
Gln Thr Leu Gln Met Arg Asn Ser Ser Cys Asp Gly Ser Arg Asp
1145 1150 1155
Tyr Ile Ile Ser Pro Val Lys Asn Ser Lys Gly Glu Phe Phe Cys
1160 1165 1170
Ser Asp Asn Asn Asp Asp Thr Thr Pro Asn Asp Ala Asp Ala Asn
1175 1180 1185
Gly Ala Phe Asn Ile Ala Arg Lys Gly Leu Trp Val Leu Asn Glu
1190 1195 1200
Ile Arg Asn Ser Glu Glu Gly Ser Lys Ile Asn Leu Ala Met Ser
1205 1210 1215
Asn Ala Gln Trp Leu Glu Tyr Ala Gln Asp Asn Thr Ile
1220 1225 1230
<210> 50
<211> 1235
<212> PRT
<213> 厌氧弧菌
<220>
<221> MISC_FEATURE
<222> (1)..(1235)
<223> Genbank WP_027407524 Cpf1
<400> 50
Met Val Ala Phe Ile Asp Glu Phe Val Gly Gln Tyr Pro Val Ser Lys
1 5 10 15
Thr Leu Arg Phe Glu Ala Arg Pro Val Pro Glu Thr Lys Lys Trp Leu
20 25 30
Glu Ser Asp Gln Cys Ser Val Leu Phe Asn Asp Gln Lys Arg Asn Glu
35 40 45
Tyr Tyr Gly Val Leu Lys Glu Leu Leu Asp Asp Tyr Tyr Arg Ala Tyr
50 55 60
Ile Glu Asp Ala Leu Thr Ser Phe Thr Leu Asp Lys Ala Leu Leu Glu
65 70 75 80
Asn Ala Tyr Asp Leu Tyr Cys Asn Arg Asp Thr Asn Ala Phe Ser Ser
85 90 95
Cys Cys Glu Lys Leu Arg Lys Asp Leu Val Lys Ala Phe Gly Asn Leu
100 105 110
Lys Asp Tyr Leu Leu Gly Ser Asp Gln Leu Lys Asp Leu Val Lys Leu
115 120 125
Lys Ala Lys Val Asp Ala Pro Ala Gly Lys Gly Lys Lys Lys Ile Glu
130 135 140
Val Asp Ser Arg Leu Ile Asn Trp Leu Asn Asn Asn Ala Lys Tyr Ser
145 150 155 160
Ala Glu Asp Arg Glu Lys Tyr Ile Lys Ala Ile Glu Ser Phe Glu Gly
165 170 175
Phe Val Thr Tyr Leu Thr Asn Tyr Lys Gln Ala Arg Glu Asn Met Phe
180 185 190
Ser Ser Glu Asp Lys Ser Thr Ala Ile Ala Phe Arg Val Ile Asp Gln
195 200 205
Asn Met Val Thr Tyr Phe Gly Asn Ile Arg Ile Tyr Glu Lys Ile Lys
210 215 220
Ala Lys Tyr Pro Glu Leu Tyr Ser Ala Leu Lys Gly Phe Glu Lys Phe
225 230 235 240
Phe Ser Pro Thr Ala Tyr Ser Glu Ile Leu Ser Gln Ser Lys Ile Asp
245 250 255
Glu Tyr Asn Tyr Gln Cys Ile Gly Arg Pro Ile Asp Asp Ala Asp Phe
260 265 270
Lys Gly Val Asn Ser Leu Ile Asn Glu Tyr Arg Gln Lys Asn Gly Ile
275 280 285
Lys Ala Arg Glu Leu Pro Val Met Ser Met Leu Tyr Lys Gln Ile Leu
290 295 300
Ser Asp Arg Asp Asn Ser Phe Met Ser Glu Val Ile Asn Arg Asn Glu
305 310 315 320
Glu Ala Ile Glu Cys Ala Lys Asn Gly Tyr Lys Val Ser Tyr Ala Leu
325 330 335
Phe Asn Glu Leu Leu Gln Leu Tyr Lys Lys Ile Phe Thr Glu Asp Asn
340 345 350
Tyr Gly Asn Ile Tyr Val Lys Thr Gln Pro Leu Thr Glu Leu Ser Gln
355 360 365
Ala Leu Phe Gly Asp Trp Ser Ile Leu Arg Asn Ala Leu Asp Asn Gly
370 375 380
Lys Tyr Asp Lys Asp Ile Ile Asn Leu Ala Glu Leu Glu Lys Tyr Phe
385 390 395 400
Ser Glu Tyr Cys Lys Val Leu Asp Ala Asp Asp Ala Ala Lys Ile Gln
405 410 415
Asp Lys Phe Asn Leu Lys Asp Tyr Phe Ile Gln Lys Asn Ala Leu Asp
420 425 430
Ala Thr Leu Pro Asp Leu Asp Lys Ile Thr Gln Tyr Lys Pro His Leu
435 440 445
Asp Ala Met Leu Gln Ala Ile Arg Lys Tyr Lys Leu Phe Ser Met Tyr
450 455 460
Asn Gly Arg Lys Lys Met Asp Val Pro Glu Asn Gly Ile Asp Phe Ser
465 470 475 480
Asn Glu Phe Asn Ala Ile Tyr Asp Lys Leu Ser Glu Phe Ser Ile Leu
485 490 495
Tyr Asp Arg Ile Arg Asn Phe Ala Thr Lys Lys Pro Tyr Ser Asp Glu
500 505 510
Lys Met Lys Leu Ser Phe Asn Met Pro Thr Met Leu Ala Gly Trp Asp
515 520 525
Tyr Asn Asn Glu Thr Ala Asn Gly Cys Phe Leu Phe Ile Lys Asp Gly
530 535 540
Lys Tyr Phe Leu Gly Val Ala Asp Ser Lys Ser Lys Asn Ile Phe Asp
545 550 555 560
Phe Lys Lys Asn Pro His Leu Leu Asp Lys Tyr Ser Ser Lys Asp Ile
565 570 575
Tyr Tyr Lys Val Lys Tyr Lys Gln Val Ser Gly Ser Ala Lys Met Leu
580 585 590
Pro Lys Val Val Phe Ala Gly Ser Asn Glu Lys Ile Phe Gly His Leu
595 600 605
Ile Ser Lys Arg Ile Leu Glu Ile Arg Glu Lys Lys Leu Tyr Thr Ala
610 615 620
Ala Ala Gly Asp Arg Lys Ala Val Ala Glu Trp Ile Asp Phe Met Lys
625 630 635 640
Ser Ala Ile Ala Ile His Pro Glu Trp Asn Glu Tyr Phe Lys Phe Lys
645 650 655
Phe Lys Asn Thr Ala Glu Tyr Asp Asn Ala Asn Lys Phe Tyr Glu Asp
660 665 670
Ile Asp Lys Gln Thr Tyr Ser Leu Glu Lys Val Glu Ile Pro Thr Glu
675 680 685
Tyr Ile Asp Glu Met Val Ser Gln His Lys Leu Tyr Leu Phe Gln Leu
690 695 700
Tyr Thr Lys Asp Phe Ser Asp Lys Lys Lys Lys Lys Gly Thr Asp Asn
705 710 715 720
Leu His Thr Met Tyr Trp His Gly Val Phe Ser Asp Glu Asn Leu Lys
725 730 735
Ala Val Thr Glu Gly Thr Gln Pro Ile Ile Lys Leu Asn Gly Glu Ala
740 745 750
Glu Met Phe Met Arg Asn Pro Ser Ile Glu Phe Gln Val Thr His Glu
755 760 765
His Asn Lys Pro Ile Ala Asn Lys Asn Pro Leu Asn Thr Lys Lys Glu
770 775 780
Ser Val Phe Asn Tyr Asp Leu Ile Lys Asp Lys Arg Tyr Thr Glu Arg
785 790 795 800
Lys Phe Tyr Phe His Cys Pro Ile Thr Leu Asn Phe Arg Ala Asp Lys
805 810 815
Pro Ile Lys Tyr Asn Glu Lys Ile Asn Arg Phe Val Glu Asn Asn Pro
820 825 830
Asp Val Cys Ile Ile Gly Ile Asp Arg Gly Glu Arg His Leu Leu Tyr
835 840 845
Tyr Thr Val Ile Asn Gln Thr Gly Asp Ile Leu Glu Gln Gly Ser Leu
850 855 860
Asn Lys Ile Ser Gly Ser Tyr Thr Asn Asp Lys Gly Glu Lys Val Asn
865 870 875 880
Lys Glu Thr Asp Tyr His Asp Leu Leu Asp Arg Lys Glu Lys Gly Lys
885 890 895
His Val Ala Gln Gln Ala Trp Glu Thr Ile Glu Asn Ile Lys Glu Leu
900 905 910
Lys Ala Gly Tyr Leu Ser Gln Val Val Tyr Lys Leu Thr Gln Leu Met
915 920 925
Leu Gln Tyr Asn Ala Val Ile Val Leu Glu Asn Leu Asn Val Gly Phe
930 935 940
Lys Arg Gly Arg Thr Lys Val Glu Lys Gln Val Tyr Gln Lys Phe Glu
945 950 955 960
Lys Ala Met Ile Asp Lys Leu Asn Tyr Leu Val Phe Lys Asp Arg Gly
965 970 975
Tyr Glu Met Asn Gly Ser Tyr Ala Lys Gly Leu Gln Leu Thr Asp Lys
980 985 990
Phe Glu Ser Phe Asp Lys Ile Gly Lys Gln Thr Gly Cys Ile Tyr Tyr
995 1000 1005
Val Ile Pro Ser Tyr Thr Ser His Ile Asp Pro Lys Thr Gly Phe
1010 1015 1020
Val Asn Leu Leu Asn Ala Lys Leu Arg Tyr Glu Asn Ile Thr Lys
1025 1030 1035
Ala Gln Asp Thr Ile Arg Lys Phe Asp Ser Ile Ser Tyr Asn Ala
1040 1045 1050
Lys Ala Asp Tyr Phe Glu Phe Ala Phe Asp Tyr Arg Ser Phe Gly
1055 1060 1065
Val Asp Met Ala Arg Asn Glu Trp Val Val Cys Thr Cys Gly Asp
1070 1075 1080
Leu Arg Trp Glu Tyr Ser Ala Lys Thr Arg Glu Thr Lys Ala Tyr
1085 1090 1095
Ser Val Thr Asp Arg Leu Lys Glu Leu Phe Lys Ala His Gly Ile
1100 1105 1110
Asp Tyr Val Gly Gly Glu Asn Leu Val Ser His Ile Thr Glu Val
1115 1120 1125
Ala Asp Lys His Phe Leu Ser Thr Leu Leu Phe Tyr Leu Arg Leu
1130 1135 1140
Val Leu Lys Met Arg Tyr Thr Val Ser Gly Thr Glu Asn Glu Asn
1145 1150 1155
Asp Phe Ile Leu Ser Pro Val Glu Tyr Ala Pro Gly Lys Phe Phe
1160 1165 1170
Asp Ser Arg Glu Ala Thr Ser Thr Glu Pro Met Asn Ala Asp Ala
1175 1180 1185
Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Leu Met Thr Ile Arg
1190 1195 1200
Gly Ile Glu Asp Gly Lys Leu His Asn Tyr Gly Lys Gly Gly Glu
1205 1210 1215
Asn Ala Ala Trp Phe Lys Phe Met Gln Asn Gln Glu Tyr Lys Asn
1220 1225 1230
Asn Gly
1235
<210> 51
<211> 1205
<212> PRT
<213> 瘤胃假丁酸弧菌
<220>
<221> MISC_FEATURE
<222> (1)..(1205)
<223> Genbank WP_028248456 Cpf1
<400> 51
Met Tyr Tyr Gln Asn Leu Thr Lys Met Tyr Pro Ile Ser Lys Thr Leu
1 5 10 15
Arg Asn Glu Leu Ile Pro Val Gly Lys Thr Leu Glu Asn Ile Arg Lys
20 25 30
Asn Gly Ile Leu Glu Ala Asp Ile Gln Arg Lys Ala Asp Tyr Glu His
35 40 45
Val Lys Lys Leu Met Asp Asn Tyr His Lys Gln Leu Ile Asn Glu Ala
50 55 60
Leu Gln Gly Val His Leu Ser Asp Leu Ser Asp Ala Tyr Asp Leu Tyr
65 70 75 80
Phe Asn Leu Ser Lys Glu Lys Asn Ser Val Asp Ala Phe Ser Lys Cys
85 90 95
Gln Asp Lys Leu Arg Lys Glu Ile Val Ser Leu Leu Lys Asn His Glu
100 105 110
Asn Phe Pro Lys Ile Gly Asn Lys Glu Ile Ile Lys Leu Leu Gln Ser
115 120 125
Leu Tyr Asp Asn Asp Thr Asp Tyr Lys Ala Leu Asp Ser Phe Ser Asn
130 135 140
Phe Tyr Thr Tyr Phe Ser Ser Tyr Asn Glu Val Arg Lys Asn Leu Tyr
145 150 155 160
Ser Asp Glu Glu Lys Ser Ser Thr Val Ala Tyr Arg Leu Ile Asn Glu
165 170 175
Asn Leu Pro Lys Phe Leu Asp Asn Ile Lys Ala Tyr Ala Ile Ala Lys
180 185 190
Lys Ala Gly Val Arg Ala Glu Gly Leu Ser Glu Glu Asp Gln Asp Cys
195 200 205
Leu Phe Ile Ile Glu Thr Phe Glu Arg Thr Leu Thr Gln Asp Gly Ile
210 215 220
Asp Asn Tyr Asn Ala Ala Ile Gly Lys Leu Asn Thr Ala Ile Asn Leu
225 230 235 240
Phe Asn Gln Gln Asn Lys Lys Gln Glu Gly Phe Arg Lys Val Pro Gln
245 250 255
Met Lys Cys Leu Tyr Lys Gln Ile Leu Ser Asp Arg Glu Glu Ala Phe
260 265 270
Ile Asp Glu Phe Ser Asp Asp Glu Asp Leu Ile Thr Asn Ile Glu Ser
275 280 285
Phe Ala Glu Asn Met Asn Val Phe Leu Asn Ser Glu Ile Ile Thr Asp
290 295 300
Phe Lys Ile Ala Leu Val Glu Ser Asp Gly Ser Leu Val Tyr Ile Lys
305 310 315 320
Asn Asp Val Ser Lys Thr Ser Phe Ser Asn Ile Val Phe Gly Ser Trp
325 330 335
Asn Ala Ile Asp Glu Lys Leu Ser Asp Glu Tyr Asp Leu Ala Asn Ser
340 345 350
Lys Lys Lys Lys Asp Glu Lys Tyr Tyr Glu Lys Arg Gln Lys Glu Leu
355 360 365
Lys Lys Asn Lys Ser Tyr Asp Leu Glu Thr Ile Ile Gly Leu Phe Asp
370 375 380
Asp Asn Ser Asp Val Ile Gly Lys Tyr Ile Glu Lys Leu Glu Ser Asp
385 390 395 400
Ile Thr Ala Ile Ala Glu Ala Lys Asn Asp Phe Asp Glu Ile Val Leu
405 410 415
Arg Lys His Asp Lys Asn Lys Ser Leu Arg Lys Asn Thr Asn Ala Val
420 425 430
Glu Ala Ile Lys Ser Tyr Leu Asp Thr Val Lys Asp Phe Glu Arg Asp
435 440 445
Ile Lys Leu Ile Asn Gly Ser Gly Gln Glu Val Glu Lys Asn Leu Val
450 455 460
Val Tyr Ala Glu Gln Glu Asn Ile Leu Ala Glu Ile Lys Asn Val Asp
465 470 475 480
Ser Leu Tyr Asn Met Ser Arg Asn Tyr Leu Thr Gln Lys Pro Phe Ser
485 490 495
Thr Glu Lys Phe Lys Leu Asn Phe Asn Arg Ala Thr Leu Leu Asn Gly
500 505 510
Trp Asp Lys Asn Lys Glu Thr Asp Asn Leu Gly Ile Leu Phe Glu Lys
515 520 525
Asp Gly Met Tyr Tyr Leu Gly Ile Met Asn Thr Lys Ala Asn Lys Ile
530 535 540
Phe Val Asn Ile Pro Lys Ala Thr Ser Asn Asp Val Tyr His Lys Val
545 550 555 560
Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met Leu Pro Lys Val Phe
565 570 575
Phe Ala Gln Ser Asn Leu Asp Tyr Tyr Lys Pro Ser Glu Glu Leu Leu
580 585 590
Ala Lys Tyr Lys Ala Gly Thr His Lys Lys Gly Asp Asn Phe Ser Leu
595 600 605
Glu Asp Cys His Ala Leu Ile Asp Phe Phe Lys Ala Ser Ile Glu Lys
610 615 620
His Pro Asp Trp Ser Ser Phe Gly Phe Glu Phe Ser Glu Thr Cys Thr
625 630 635 640
Tyr Glu Asp Leu Ser Gly Phe Tyr Arg Glu Val Glu Lys Gln Gly Tyr
645 650 655
Lys Ile Thr Tyr Thr Asp Val Asp Ala Asp Tyr Ile Thr Ser Leu Val
660 665 670
Glu Arg Asp Glu Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser
675 680 685
Pro Tyr Ser Lys Gly Asn Leu Asn Leu His Thr Ile Tyr Leu Gln Met
690 695 700
Leu Phe Asp Gln Arg Asn Leu Asn Asn Val Val Tyr Lys Leu Asn Gly
705 710 715 720
Glu Ala Glu Val Phe Tyr Arg Pro Ala Ser Ile Asn Asp Glu Glu Val
725 730 735
Ile Ile His Lys Ala Gly Glu Glu Ile Lys Asn Lys Asn Ser Lys Arg
740 745 750
Ala Val Asp Lys Pro Thr Ser Lys Phe Gly Tyr Asp Ile Ile Lys Asp
755 760 765
Arg Arg Tyr Ser Lys Asp Lys Phe Met Leu His Ile Pro Val Thr Met
770 775 780
Asn Phe Gly Val Asp Glu Thr Arg Arg Phe Asn Asp Val Val Asn Asp
785 790 795 800
Ala Leu Arg Asn Asp Glu Lys Val Arg Val Ile Gly Ile Asp Arg Gly
805 810 815
Glu Arg Asn Leu Leu Tyr Val Val Val Val Asp Thr Asp Gly Thr Ile
820 825 830
Leu Glu Gln Ile Ser Leu Asn Ser Ile Ile Asn Asn Glu Tyr Ser Ile
835 840 845
Glu Thr Asp Tyr His Lys Leu Leu Asp Glu Lys Glu Gly Asp Arg Asp
850 855 860
Arg Ala Arg Lys Asn Trp Thr Thr Ile Glu Asn Ile Lys Glu Leu Lys
865 870 875 880
Glu Gly Tyr Leu Ser Gln Val Val Asn Val Ile Ala Lys Leu Val Leu
885 890 895
Lys Tyr Asn Ala Ile Ile Cys Leu Glu Asp Leu Asn Phe Gly Phe Lys
900 905 910
Arg Gly Arg Gln Lys Val Glu Lys Gln Val Tyr Gln Lys Phe Glu Lys
915 920 925
Met Leu Ile Asp Lys Leu Asn Tyr Leu Val Ile Asp Lys Ser Arg Lys
930 935 940
Gln Asp Lys Pro Glu Glu Phe Gly Gly Ala Leu Asn Ala Leu Gln Leu
945 950 955 960
Thr Ser Lys Phe Thr Ser Phe Lys Asp Met Gly Lys Gln Thr Gly Ile
965 970 975
Ile Tyr Tyr Val Pro Ala Tyr Leu Thr Ser Lys Ile Asp Pro Thr Thr
980 985 990
Gly Phe Ala Asn Leu Phe Tyr Val Lys Tyr Glu Asn Val Glu Lys Ala
995 1000 1005
Lys Glu Phe Phe Ser Arg Phe Asp Ser Ile Ser Tyr Asn Asn Glu
1010 1015 1020
Ser Gly Tyr Phe Glu Phe Ala Phe Asp Tyr Lys Lys Phe Thr Asp
1025 1030 1035
Arg Ala Cys Gly Ala Arg Ser Gln Trp Thr Val Cys Thr Tyr Gly
1040 1045 1050
Glu Arg Ile Ile Lys Phe Arg Asn Thr Glu Lys Asn Asn Ser Phe
1055 1060 1065
Asp Asp Lys Thr Ile Val Leu Ser Glu Glu Phe Lys Glu Leu Phe
1070 1075 1080
Ser Ile Tyr Gly Ile Ser Tyr Glu Asp Gly Ala Glu Leu Lys Asn
1085 1090 1095
Lys Ile Met Ser Val Asp Glu Ala Asp Phe Phe Arg Ser Leu Thr
1100 1105 1110
Arg Leu Phe Gln Gln Thr Met Gln Met Arg Asn Ser Ser Asn Asp
1115 1120 1125
Val Thr Arg Asp Tyr Ile Ile Ser Pro Ile Met Asn Asp Arg Gly
1130 1135 1140
Glu Phe Phe Asn Ser Glu Ala Cys Asp Ala Ser Lys Pro Lys Asp
1145 1150 1155
Ala Asp Ala Asn Gly Ala Phe Asn Ile Ala Arg Lys Gly Leu Trp
1160 1165 1170
Val Leu Glu Gln Ile Arg Asn Thr Pro Ser Gly Asp Lys Leu Asn
1175 1180 1185
Leu Ala Met Ser Asn Ala Glu Trp Leu Glu Tyr Ala Gln Arg Asn
1190 1195 1200
Gln Ile
1205
<210> 52
<211> 1154
<212> PRT
<213> 产丁酸菌
<220>
<221> MISC_FEATURE
<222> (1)..(1154)
<223> Genbank WP_028830240 Cpf1
<400> 52
Met Glu Asn Phe Lys Asn Leu Tyr Pro Ile Asn Lys Thr Leu Arg Phe
1 5 10 15
Glu Leu Arg Pro Tyr Gly Lys Thr Leu Glu Asn Phe Lys Lys Ser Gly
20 25 30
Leu Leu Glu Lys Asp Ala Phe Lys Ala Asn Ser Arg Arg Ser Met Gln
35 40 45
Ala Ile Ile Asp Glu Lys Phe Lys Glu Thr Ile Glu Glu Arg Leu Lys
50 55 60
Tyr Thr Glu Phe Ser Glu Cys Asp Leu Gly Asn Met Thr Ser Lys Asp
65 70 75 80
Lys Lys Ile Thr Asp Lys Ala Ala Thr Asn Leu Lys Lys Gln Val Ile
85 90 95
Leu Ser Phe Asp Asp Glu Ile Phe Asn Asn Tyr Leu Lys Pro Asp Lys
100 105 110
Asn Ile Asp Ala Leu Phe Lys Asn Asp Pro Ser Asn Pro Val Ile Ser
115 120 125
Thr Phe Lys Gly Phe Thr Thr Tyr Phe Val Asn Phe Phe Glu Ile Arg
130 135 140
Lys His Ile Phe Lys Gly Glu Ser Ser Gly Ser Met Ala Tyr Arg Ile
145 150 155 160
Ile Asp Glu Asn Leu Thr Thr Tyr Leu Asn Asn Ile Glu Lys Ile Lys
165 170 175
Lys Leu Pro Glu Glu Leu Lys Ser Gln Leu Glu Gly Ile Asp Gln Ile
180 185 190
Asp Lys Leu Asn Asn Tyr Asn Glu Phe Ile Thr Gln Ser Gly Ile Thr
195 200 205
His Tyr Asn Glu Ile Ile Gly Gly Ile Ser Lys Ser Glu Asn Val Lys
210 215 220
Ile Gln Gly Ile Asn Glu Gly Ile Asn Leu Tyr Cys Gln Lys Asn Lys
225 230 235 240
Val Lys Leu Pro Arg Leu Thr Pro Leu Tyr Lys Met Ile Leu Ser Asp
245 250 255
Arg Val Ser Asn Ser Phe Val Leu Asp Thr Ile Glu Asn Asp Thr Glu
260 265 270
Leu Ile Glu Met Ile Ser Asp Leu Ile Asn Lys Thr Glu Ile Ser Gln
275 280 285
Asp Val Ile Met Ser Asp Ile Gln Asn Ile Phe Ile Lys Tyr Lys Gln
290 295 300
Leu Gly Asn Leu Pro Gly Ile Ser Tyr Ser Ser Ile Val Asn Ala Ile
305 310 315 320
Cys Ser Asp Tyr Asp Asn Asn Phe Gly Asp Gly Lys Arg Lys Lys Ser
325 330 335
Tyr Glu Asn Asp Arg Lys Lys His Leu Glu Thr Asn Val Tyr Ser Ile
340 345 350
Asn Tyr Ile Ser Glu Leu Leu Thr Asp Thr Asp Val Ser Ser Asn Ile
355 360 365
Lys Met Arg Tyr Lys Glu Leu Glu Gln Asn Tyr Gln Val Cys Lys Glu
370 375 380
Asn Phe Asn Ala Thr Asn Trp Met Asn Ile Lys Asn Ile Lys Gln Ser
385 390 395 400
Glu Lys Thr Asn Leu Ile Lys Asp Leu Leu Asp Ile Leu Lys Ser Ile
405 410 415
Gln Arg Phe Tyr Asp Leu Phe Asp Ile Val Asp Glu Asp Lys Asn Pro
420 425 430
Ser Ala Glu Phe Tyr Thr Trp Leu Ser Lys Asn Ala Glu Lys Leu Asp
435 440 445
Phe Glu Phe Asn Ser Val Tyr Asn Lys Ser Arg Asn Tyr Leu Thr Arg
450 455 460
Lys Gln Tyr Ser Asp Lys Lys Ile Lys Leu Asn Phe Asp Ser Pro Thr
465 470 475 480
Leu Ala Lys Gly Trp Asp Ala Asn Lys Glu Ile Asp Asn Ser Thr Ile
485 490 495
Ile Met Arg Lys Phe Asn Asn Asp Arg Gly Asp Tyr Asp Tyr Phe Leu
500 505 510
Gly Ile Trp Asn Lys Ser Thr Pro Ala Asn Glu Lys Ile Ile Pro Leu
515 520 525
Glu Asp Asn Gly Leu Phe Glu Lys Met Gln Tyr Lys Leu Tyr Pro Asp
530 535 540
Pro Ser Lys Met Leu Pro Lys Gln Phe Leu Ser Lys Ile Trp Lys Ala
545 550 555 560
Lys His Pro Thr Thr Pro Glu Phe Asp Lys Lys Tyr Lys Glu Gly Arg
565 570 575
His Lys Lys Gly Pro Asp Phe Glu Lys Glu Phe Leu His Glu Leu Ile
580 585 590
Asp Cys Phe Lys His Gly Leu Val Asn His Asp Glu Lys Tyr Gln Asp
595 600 605
Val Phe Gly Phe Asn Leu Arg Asn Thr Glu Asp Tyr Asn Ser Tyr Thr
610 615 620
Glu Phe Leu Glu Asp Val Glu Arg Cys Asn Tyr Asn Leu Ser Phe Asn
625 630 635 640
Lys Ile Ala Asp Thr Ser Asn Leu Ile Asn Asp Gly Lys Leu Tyr Val
645 650 655
Phe Gln Ile Trp Ser Lys Asp Phe Ser Ile Asp Ser Lys Gly Thr Lys
660 665 670
Asn Leu Asn Thr Ile Tyr Phe Glu Ser Leu Phe Ser Glu Glu Asn Met
675 680 685
Ile Glu Lys Met Phe Lys Leu Ser Gly Glu Ala Glu Ile Phe Tyr Arg
690 695 700
Pro Ala Ser Leu Asn Tyr Cys Glu Asp Ile Ile Lys Lys Gly His His
705 710 715 720
His Ala Glu Leu Lys Asp Lys Phe Asp Tyr Pro Ile Ile Lys Asp Lys
725 730 735
Arg Tyr Ser Gln Asp Lys Phe Phe Phe His Val Pro Met Val Ile Asn
740 745 750
Tyr Lys Ser Glu Lys Leu Asn Ser Lys Ser Leu Asn Asn Arg Thr Asn
755 760 765
Glu Asn Leu Gly Gln Phe Thr His Ile Ile Gly Ile Asp Arg Gly Glu
770 775 780
Arg His Leu Ile Tyr Leu Thr Val Val Asp Val Ser Thr Gly Glu Ile
785 790 795 800
Val Glu Gln Lys His Leu Asp Glu Ile Ile Asn Thr Asp Thr Lys Gly
805 810 815
Val Glu His Lys Thr His Tyr Leu Asn Lys Leu Glu Glu Lys Ser Lys
820 825 830
Thr Arg Asp Asn Glu Arg Lys Ser Trp Glu Ala Ile Glu Thr Ile Lys
835 840 845
Glu Leu Lys Glu Gly Tyr Ile Ser His Val Ile Asn Glu Ile Gln Lys
850 855 860
Leu Gln Glu Lys Tyr Asn Ala Leu Ile Val Met Glu Asn Leu Asn Tyr
865 870 875 880
Gly Phe Lys Asn Ser Arg Ile Lys Val Glu Lys Gln Val Tyr Gln Lys
885 890 895
Phe Glu Thr Ala Leu Ile Lys Lys Phe Asn Tyr Ile Ile Asp Lys Lys
900 905 910
Asp Pro Glu Thr Tyr Ile His Gly Tyr Gln Leu Thr Asn Pro Ile Thr
915 920 925
Thr Leu Asp Lys Ile Gly Asn Gln Ser Gly Ile Val Leu Tyr Ile Pro
930 935 940
Ala Trp Asn Thr Ser Lys Ile Asp Pro Val Thr Gly Phe Val Asn Leu
945 950 955 960
Leu Tyr Ala Asp Asp Leu Lys Tyr Lys Asn Gln Glu Gln Ala Lys Ser
965 970 975
Phe Ile Gln Lys Ile Asp Asn Ile Tyr Phe Glu Asn Gly Glu Phe Lys
980 985 990
Phe Asp Ile Asp Phe Ser Lys Trp Asn Asn Arg Tyr Ser Ile Ser Lys
995 1000 1005
Thr Lys Trp Thr Leu Thr Ser Tyr Gly Thr Arg Ile Gln Thr Phe
1010 1015 1020
Arg Asn Pro Gln Lys Asn Asn Lys Trp Asp Ser Ala Glu Tyr Asp
1025 1030 1035
Leu Thr Glu Glu Phe Lys Leu Ile Leu Asn Ile Asp Gly Thr Leu
1040 1045 1050
Lys Ser Gln Asp Val Glu Thr Tyr Lys Lys Phe Met Ser Leu Phe
1055 1060 1065
Lys Leu Met Leu Gln Leu Arg Asn Ser Val Thr Gly Thr Asp Ile
1070 1075 1080
Asp Tyr Met Ile Ser Pro Val Thr Asp Lys Thr Gly Thr His Phe
1085 1090 1095
Asp Ser Arg Glu Asn Ile Lys Asn Leu Pro Ala Asp Ala Asp Ala
1100 1105 1110
Asn Gly Ala Tyr Asn Ile Ala Arg Lys Gly Ile Met Ala Ile Glu
1115 1120 1125
Asn Ile Met Asn Gly Ile Ser Asp Pro Leu Lys Ile Ser Asn Glu
1130 1135 1140
Asp Tyr Leu Lys Tyr Ile Gln Asn Gln Gln Glu
1145 1150
<210> 53
<211> 1206
<212> PRT
<213> 丁酸弧菌
<220>
<221> MISC_FEATURE
<222> (1)..(1206)
<223> Genbank WP_035798880 Cpf1
<400> 53
Met Tyr Tyr Gln Asn Leu Thr Lys Lys Tyr Pro Val Ser Lys Thr Ile
1 5 10 15
Arg Asn Glu Leu Ile Pro Ile Gly Lys Thr Leu Glu Asn Ile Arg Lys
20 25 30
Asn Asn Ile Leu Glu Ser Asp Val Lys Arg Lys Gln Asp Tyr Glu His
35 40 45
Val Lys Gly Ile Met Asp Glu Tyr His Lys Gln Leu Ile Asn Glu Ala
50 55 60
Leu Asp Asn Tyr Met Leu Pro Ser Leu Asn Gln Ala Ala Glu Ile Tyr
65 70 75 80
Leu Lys Lys His Val Asp Val Glu Asp Arg Glu Glu Phe Lys Lys Thr
85 90 95
Gln Asp Leu Leu Arg Arg Glu Val Thr Gly Arg Leu Lys Glu His Glu
100 105 110
Asn Tyr Thr Lys Ile Gly Lys Lys Asp Ile Leu Asp Leu Leu Glu Lys
115 120 125
Leu Pro Ser Ile Ser Glu Glu Asp Tyr Asn Ala Leu Glu Ser Phe Arg
130 135 140
Asn Phe Tyr Thr Tyr Phe Thr Ser Tyr Asn Lys Val Arg Glu Asn Leu
145 150 155 160
Tyr Ser Asp Glu Glu Lys Ser Ser Thr Val Ala Tyr Arg Leu Ile Asn
165 170 175
Glu Asn Leu Pro Lys Phe Leu Asp Asn Ile Lys Ser Tyr Ala Phe Val
180 185 190
Lys Ala Ala Gly Val Leu Ala Asp Cys Ile Glu Glu Glu Glu Gln Asp
195 200 205
Ala Leu Phe Met Val Glu Thr Phe Asn Met Thr Leu Thr Gln Glu Gly
210 215 220
Ile Asp Met Tyr Asn Tyr Gln Ile Gly Lys Val Asn Ser Ala Ile Asn
225 230 235 240
Leu Tyr Asn Gln Lys Asn His Lys Val Glu Glu Phe Lys Lys Ile Pro
245 250 255
Lys Met Lys Val Leu Tyr Lys Gln Ile Leu Ser Asp Arg Glu Glu Val
260 265 270
Phe Ile Gly Glu Phe Lys Asp Asp Glu Thr Leu Leu Ser Ser Ile Gly
275 280 285
Ala Tyr Gly Asn Val Leu Met Thr Tyr Leu Lys Ser Glu Lys Ile Asn
290 295 300
Ile Phe Phe Asp Ala Leu Arg Glu Ser Glu Gly Lys Asn Val Tyr Val
305 310 315 320
Lys Asn Asp Leu Ser Lys Thr Thr Met Ser Asn Ile Val Phe Gly Ser
325 330 335
Trp Ser Ala Phe Asp Glu Leu Leu Asn Gln Glu Tyr Asp Leu Ala Asn
340 345 350
Glu Asn Lys Lys Lys Asp Asp Lys Tyr Phe Glu Lys Arg Gln Lys Glu
355 360 365
Leu Lys Lys Asn Lys Ser Tyr Thr Leu Glu Gln Met Ser Asn Leu Ser
370 375 380
Lys Glu Asp Ile Ser Pro Ile Glu Asn Tyr Ile Glu Arg Ile Ser Glu
385 390 395 400
Asp Ile Glu Lys Ile Cys Ile Tyr Asn Gly Glu Phe Glu Lys Ile Val
405 410 415
Val Asn Glu His Asp Ser Ser Arg Lys Leu Ser Lys Asn Ile Lys Ala
420 425 430
Val Lys Val Ile Lys Asp Tyr Leu Asp Ser Ile Lys Glu Leu Glu His
435 440 445
Asp Ile Lys Leu Ile Asn Gly Ser Gly Gln Glu Leu Glu Lys Asn Leu
450 455 460
Val Val Tyr Val Gly Gln Glu Glu Ala Leu Glu Gln Leu Arg Pro Val
465 470 475 480
Asp Ser Leu Tyr Asn Leu Thr Arg Asn Tyr Leu Thr Lys Lys Pro Phe
485 490 495
Ser Thr Glu Lys Val Lys Leu Asn Phe Asn Lys Ser Thr Leu Leu Asn
500 505 510
Gly Trp Asp Lys Asn Lys Glu Thr Asp Asn Leu Gly Ile Leu Phe Phe
515 520 525
Lys Asp Gly Lys Tyr Tyr Leu Gly Ile Met Asn Thr Thr Ala Asn Lys
530 535 540
Ala Phe Val Asn Pro Pro Ala Ala Lys Thr Glu Asn Val Phe Lys Lys
545 550 555 560
Val Asp Tyr Lys Leu Leu Pro Gly Ser Asn Lys Met Leu Pro Lys Val
565 570 575
Phe Phe Ala Lys Ser Asn Ile Gly Tyr Tyr Asn Pro Ser Thr Glu Leu
580 585 590
Tyr Ser Asn Tyr Lys Lys Gly Thr His Lys Lys Gly Pro Ser Phe Ser
595 600 605
Ile Asp Asp Cys His Asn Leu Ile Asp Phe Phe Lys Glu Ser Ile Lys
610 615 620
Lys His Glu Asp Trp Ser Lys Phe Gly Phe Glu Phe Ser Asp Thr Ala
625 630 635 640
Asp Tyr Arg Asp Ile Ser Glu Phe Tyr Arg Glu Val Glu Lys Gln Gly
645 650 655
Tyr Lys Leu Thr Phe Thr Asp Ile Asp Glu Ser Tyr Ile Asn Asp Leu
660 665 670
Ile Glu Lys Asn Glu Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe
675 680 685
Ser Glu Tyr Ser Lys Gly Lys Leu Asn Leu His Thr Leu Tyr Phe Met
690 695 700
Met Leu Phe Asp Gln Arg Asn Leu Asp Asn Val Val Tyr Lys Leu Asn
705 710 715 720
Gly Glu Ala Glu Val Phe Tyr Arg Pro Ala Ser Ile Ala Glu Asn Glu
725 730 735
Leu Val Ile His Lys Ala Gly Glu Gly Ile Lys Asn Lys Asn Pro Asn
740 745 750
Arg Ala Lys Val Lys Glu Thr Ser Thr Phe Ser Tyr Asp Ile Val Lys
755 760 765
Asp Lys Arg Tyr Ser Lys Tyr Lys Phe Thr Leu His Ile Pro Ile Thr
770 775 780
Met Asn Phe Gly Val Asp Glu Val Arg Arg Phe Asn Asp Val Ile Asn
785 790 795 800
Asn Ala Leu Arg Thr Asp Asp Asn Val Asn Val Ile Gly Ile Asp Arg
805 810 815
Gly Glu Arg Asn Leu Leu Tyr Val Val Val Ile Asn Ser Glu Gly Lys
820 825 830
Ile Leu Glu Gln Ile Ser Leu Asn Ser Ile Ile Asn Lys Glu Tyr Asp
835 840 845
Ile Glu Thr Asn Tyr His Ala Leu Leu Asp Glu Arg Glu Asp Asp Arg
850 855 860
Asn Lys Ala Arg Lys Asp Trp Asn Thr Ile Glu Asn Ile Lys Glu Leu
865 870 875 880
Lys Thr Gly Tyr Leu Ser Gln Val Val Asn Val Val Ala Lys Leu Val
885 890 895
Leu Lys Tyr Asn Ala Ile Ile Cys Leu Glu Asp Leu Asn Phe Gly Phe
900 905 910
Lys Arg Gly Arg Gln Lys Val Glu Lys Gln Val Tyr Gln Lys Phe Glu
915 920 925
Lys Met Leu Ile Glu Lys Leu Asn Tyr Leu Val Ile Asp Lys Ser Arg
930 935 940
Glu Gln Val Ser Pro Glu Lys Met Gly Gly Ala Leu Asn Ala Leu Gln
945 950 955 960
Leu Thr Ser Lys Phe Lys Ser Phe Ala Glu Leu Gly Lys Gln Ser Gly
965 970 975
Ile Ile Tyr Tyr Val Pro Ala Tyr Leu Thr Ser Lys Ile Asp Pro Thr
980 985 990
Thr Gly Phe Val Asn Leu Phe Tyr Ile Lys Tyr Glu Asn Ile Glu Lys
995 1000 1005
Ala Lys Gln Phe Phe Asp Gly Phe Asp Phe Ile Arg Phe Asn Lys
1010 1015 1020
Lys Asp Asp Met Phe Glu Phe Ser Phe Asp Tyr Lys Ser Phe Thr
1025 1030 1035
Gln Lys Ala Cys Gly Ile Arg Ser Lys Trp Ile Val Tyr Thr Asn
1040 1045 1050
Gly Glu Arg Ile Ile Lys Tyr Pro Asn Pro Glu Lys Asn Asn Leu
1055 1060 1065
Phe Asp Glu Lys Val Ile Asn Val Thr Asp Glu Ile Lys Gly Leu
1070 1075 1080
Phe Lys Gln Tyr Arg Ile Pro Tyr Glu Asn Gly Glu Asp Ile Lys
1085 1090 1095
Glu Ile Ile Ile Ser Lys Ala Glu Ala Asp Phe Tyr Lys Arg Leu
1100 1105 1110
Phe Arg Leu Leu His Gln Thr Leu Gln Met Arg Asn Ser Thr Ser
1115 1120 1125
Asp Gly Thr Arg Asp Tyr Ile Ile Ser Pro Val Lys Asn Asp Arg
1130 1135 1140
Gly Glu Phe Phe Cys Ser Glu Phe Ser Glu Gly Thr Met Pro Lys
1145 1150 1155
Asp Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala Arg Lys Gly Leu
1160 1165 1170
Trp Val Leu Glu Gln Ile Arg Gln Lys Asp Glu Gly Glu Lys Val
1175 1180 1185
Asn Leu Ser Met Thr Asn Ala Glu Trp Leu Lys Tyr Ala Gln Leu
1190 1195 1200
His Leu Leu
1205
<210> 54
<211> 1264
<212> PRT
<213> 卡普拉莫拉氏菌
<220>
<221> MISC_FEATURE
<222> (1)..(1264)
<223> Genbank WP_036388671 Cpf1
<400> 54
Met Leu Phe Gln Asp Phe Thr His Leu Tyr Pro Leu Ser Lys Thr Met
1 5 10 15
Arg Phe Glu Leu Lys Pro Ile Gly Lys Thr Leu Glu His Ile His Ala
20 25 30
Lys Asn Phe Leu Ser Gln Asp Glu Thr Met Ala Asp Met Tyr Gln Lys
35 40 45
Val Lys Ala Ile Leu Asp Asp Tyr His Arg Asp Phe Ile Ala Asp Met
50 55 60
Met Gly Glu Val Lys Leu Thr Lys Leu Ala Glu Phe Tyr Asp Val Tyr
65 70 75 80
Leu Lys Phe Arg Lys Asn Pro Lys Asp Asp Gly Leu Gln Lys Gln Leu
85 90 95
Lys Asp Leu Gln Ala Val Leu Arg Lys Glu Ile Val Lys Pro Ile Gly
100 105 110
Asn Gly Gly Lys Tyr Lys Ala Gly Tyr Asp Arg Leu Phe Gly Ala Lys
115 120 125
Leu Phe Lys Asp Gly Lys Glu Leu Gly Asp Leu Ala Lys Phe Val Ile
130 135 140
Ala Gln Glu Gly Glu Ser Ser Pro Lys Leu Ala His Leu Ala His Phe
145 150 155 160
Glu Lys Phe Ser Thr Tyr Phe Thr Gly Phe His Asp Asn Arg Lys Asn
165 170 175
Met Tyr Ser Asp Glu Asp Lys His Thr Ala Ile Thr Tyr Arg Leu Ile
180 185 190
His Glu Asn Leu Pro Arg Phe Ile Asp Asn Leu Gln Ile Leu Ala Thr
195 200 205
Ile Lys Gln Lys His Ser Ala Leu Tyr Asp Gln Ile Ile Asn Glu Leu
210 215 220
Thr Ala Ser Gly Leu Asp Val Ser Leu Ala Ser His Leu Asp Gly Tyr
225 230 235 240
His Lys Leu Leu Thr Gln Glu Gly Ile Thr Ala Tyr Asn Thr Leu Leu
245 250 255
Gly Gly Ile Ser Gly Glu Ala Gly Ser Arg Lys Ile Gln Gly Ile Asn
260 265 270
Glu Leu Ile Asn Ser His His Asn Gln His Cys His Lys Ser Glu Arg
275 280 285
Ile Ala Lys Leu Arg Pro Leu His Lys Gln Ile Leu Ser Asp Gly Met
290 295 300
Gly Val Ser Phe Leu Pro Ser Lys Phe Ala Asp Asp Ser Glu Met Cys
305 310 315 320
Gln Ala Val Asn Glu Phe Tyr Arg His Tyr Ala Asp Val Phe Ala Lys
325 330 335
Val Gln Ser Leu Phe Asp Gly Phe Asp Asp His Gln Lys Asp Gly Ile
340 345 350
Tyr Val Glu His Lys Asn Leu Asn Glu Leu Ser Lys Gln Ala Phe Gly
355 360 365
Asp Phe Ala Leu Leu Gly Arg Val Leu Asp Gly Tyr Tyr Val Asp Val
370 375 380
Val Asn Pro Glu Phe Asn Glu Arg Phe Ala Lys Ala Lys Thr Asp Asn
385 390 395 400
Ala Lys Ala Lys Leu Thr Lys Glu Lys Asp Lys Phe Ile Lys Gly Val
405 410 415
His Ser Leu Ala Ser Leu Glu Gln Ala Ile Glu His Tyr Thr Ala Arg
420 425 430
His Asp Asp Glu Ser Val Gln Ala Gly Lys Leu Gly Gln Tyr Phe Lys
435 440 445
His Gly Leu Ala Gly Val Asp Asn Pro Ile Gln Lys Ile His Asn Asn
450 455 460
His Ser Thr Ile Lys Gly Phe Leu Glu Arg Glu Arg Pro Ala Gly Glu
465 470 475 480
Arg Ala Leu Pro Lys Ile Lys Ser Gly Lys Asn Pro Glu Met Thr Gln
485 490 495
Leu Arg Gln Leu Lys Glu Leu Leu Asp Asn Ala Leu Asn Val Ala His
500 505 510
Phe Ala Lys Leu Leu Thr Thr Lys Thr Thr Leu Asp Asn Gln Asp Gly
515 520 525
Asn Phe Tyr Gly Glu Phe Gly Ala Leu Tyr Asp Glu Leu Ala Lys Ile
530 535 540
Pro Thr Leu Tyr Asn Lys Val Arg Asp Tyr Leu Ser Gln Lys Pro Phe
545 550 555 560
Ser Thr Glu Lys Tyr Lys Leu Asn Phe Gly Asn Pro Thr Leu Leu Asn
565 570 575
Gly Trp Asp Leu Asn Lys Glu Lys Asp Asn Phe Gly Ile Ile Leu Gln
580 585 590
Lys Asp Gly Cys Tyr Tyr Leu Ala Leu Leu Asp Lys Ala His Lys Lys
595 600 605
Val Phe Asp Asn Ala Pro Asn Thr Gly Lys Asn Val Tyr Gln Lys Met
610 615 620
Ile Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met Leu Pro Lys Val Phe
625 630 635 640
Phe Ala Lys Ser Asn Leu Asp Tyr Tyr Asn Pro Ser Ala Glu Leu Leu
645 650 655
Asp Lys Tyr Ala Gln Gly Thr His Lys Lys Gly Asn Asn Phe Asn Leu
660 665 670
Lys Asp Cys His Ala Leu Ile Asp Phe Phe Lys Ala Gly Ile Asn Lys
675 680 685
His Pro Glu Trp Gln His Phe Gly Phe Lys Phe Ser Pro Thr Ser Ser
690 695 700
Tyr Gln Asp Leu Ser Asp Phe Tyr Arg Glu Val Glu Pro Gln Gly Tyr
705 710 715 720
Gln Val Lys Phe Val Asp Ile Asn Ala Asp Tyr Ile Asn Glu Leu Val
725 730 735
Glu Gln Gly Gln Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser
740 745 750
Pro Lys Ala His Gly Lys Pro Asn Leu His Thr Leu Tyr Phe Lys Ala
755 760 765
Leu Phe Ser Lys Asp Asn Leu Ala Asn Pro Ile Tyr Lys Leu Asn Gly
770 775 780
Glu Ala Gln Ile Phe Tyr Arg Lys Ala Ser Leu Asp Met Asn Glu Thr
785 790 795 800
Thr Ile His Arg Ala Gly Glu Val Leu Glu Asn Lys Asn Pro Asp Asn
805 810 815
Pro Lys Lys Arg Gln Phe Val Tyr Asp Ile Ile Lys Asp Lys Arg Tyr
820 825 830
Thr Gln Asp Lys Phe Met Leu His Val Pro Ile Thr Met Asn Phe Gly
835 840 845
Val Gln Gly Met Thr Ile Lys Glu Phe Asn Lys Lys Val Asn Gln Ser
850 855 860
Ile Gln Gln Tyr Asp Glu Val Asn Val Ile Gly Ile Asp Arg Gly Glu
865 870 875 880
Arg His Leu Leu Tyr Leu Thr Val Ile Asn Ser Lys Gly Glu Ile Leu
885 890 895
Glu Gln Arg Ser Leu Asn Asp Ile Thr Thr Ala Ser Ala Asn Gly Thr
900 905 910
Gln Met Thr Thr Pro Tyr His Lys Ile Leu Asp Lys Arg Glu Ile Glu
915 920 925
Arg Leu Asn Ala Arg Val Gly Trp Gly Glu Ile Glu Thr Ile Lys Glu
930 935 940
Leu Lys Ser Gly Tyr Leu Ser His Val Val His Gln Ile Ser Gln Leu
945 950 955 960
Met Leu Lys Tyr Asn Ala Ile Val Val Leu Glu Asp Leu Asn Phe Gly
965 970 975
Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Ile Tyr Gln Asn Phe
980 985 990
Glu Asn Ala Leu Ile Lys Lys Leu Asn His Leu Val Leu Lys Asp Glu
995 1000 1005
Ala Asp Asp Glu Ile Gly Ser Tyr Lys Asn Ala Leu Gln Leu Thr
1010 1015 1020
Asn Asn Phe Thr Asp Leu Lys Ser Ile Gly Lys Gln Thr Gly Phe
1025 1030 1035
Leu Phe Tyr Val Pro Ala Trp Asn Thr Ser Lys Ile Asp Pro Glu
1040 1045 1050
Thr Gly Phe Val Asp Leu Leu Lys Pro Arg Tyr Glu Asn Ile Ala
1055 1060 1065
Gln Ser Gln Ala Phe Phe Gly Lys Phe Asp Lys Ile Cys Tyr Asn
1070 1075 1080
Ala Asp Lys Asp Tyr Phe Glu Phe His Ile Asp Tyr Ala Lys Phe
1085 1090 1095
Thr Asp Lys Ala Lys Asn Ser Arg Gln Ile Trp Lys Ile Cys Ser
1100 1105 1110
His Gly Asp Lys Arg Tyr Val Tyr Asp Lys Thr Ala Asn Gln Asn
1115 1120 1125
Lys Gly Ala Thr Lys Gly Ile Asn Val Asn Asp Glu Leu Lys Ser
1130 1135 1140
Leu Phe Ala Arg His His Ile Asn Asp Lys Gln Pro Asn Leu Val
1145 1150 1155
Met Asp Ile Cys Gln Asn Asn Asp Lys Glu Phe His Lys Ser Leu
1160 1165 1170
Ile Tyr Leu Leu Lys Thr Leu Leu Ala Leu Arg Tyr Ser Asn Ala
1175 1180 1185
Ser Ser Asp Glu Asp Phe Ile Leu Ser Pro Val Ala Asn Asp Glu
1190 1195 1200
Gly Met Phe Phe Asn Ser Ala Leu Ala Asp Asp Thr Gln Pro Gln
1205 1210 1215
Asn Ala Asp Ala Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Leu
1220 1225 1230
Trp Val Leu Glu Gln Ile Lys Asn Ser Asp Asp Leu Asn Lys Val
1235 1240 1245
Lys Leu Ala Ile Asp Asn Gln Thr Trp Leu Asn Phe Ala Gln Asn
1250 1255 1260
Arg
<210> 55
<211> 1259
<212> PRT
<213> 琼斯氏共生菌
<220>
<221> MISC_FEATURE
<222> (1)..(1259)
<223> Genbank WP_037975888 Cpf1
<400> 55
Met Ala Asn Ser Leu Lys Asp Phe Thr Asn Ile Tyr Gln Leu Ser Lys
1 5 10 15
Thr Leu Arg Phe Glu Leu Lys Pro Ile Gly Lys Thr Glu Glu His Ile
20 25 30
Asn Arg Lys Leu Ile Ile Met His Asp Glu Lys Arg Gly Glu Asp Tyr
35 40 45
Lys Ser Val Thr Lys Leu Ile Asp Asp Tyr His Arg Lys Phe Ile His
50 55 60
Glu Thr Leu Asp Pro Ala His Phe Asp Trp Asn Pro Leu Ala Glu Ala
65 70 75 80
Leu Ile Gln Ser Gly Ser Lys Asn Asn Lys Ala Leu Pro Ala Glu Gln
85 90 95
Lys Glu Met Arg Glu Lys Ile Ile Ser Met Phe Thr Ser Gln Ala Val
100 105 110
Tyr Lys Lys Leu Phe Lys Lys Glu Leu Phe Ser Glu Leu Leu Pro Glu
115 120 125
Met Ile Lys Ser Glu Leu Val Ser Asp Leu Glu Lys Gln Ala Gln Leu
130 135 140
Asp Ala Val Lys Ser Phe Asp Lys Phe Ser Thr Tyr Phe Thr Gly Phe
145 150 155 160
His Glu Asn Arg Lys Asn Ile Tyr Ser Lys Lys Asp Thr Ser Thr Ser
165 170 175
Ile Ala Phe Arg Ile Val His Gln Asn Phe Pro Lys Phe Leu Ala Asn
180 185 190
Val Arg Ala Tyr Thr Leu Ile Lys Glu Arg Ala Pro Glu Val Ile Asp
195 200 205
Lys Ala Gln Lys Glu Leu Ser Gly Ile Leu Gly Gly Lys Thr Leu Asp
210 215 220
Asp Ile Phe Ser Ile Glu Ser Phe Asn Asn Val Leu Thr Gln Asp Lys
225 230 235 240
Ile Asp Tyr Tyr Asn Gln Ile Ile Gly Gly Val Ser Gly Lys Ala Gly
245 250 255
Asp Lys Lys Leu Arg Gly Val Asn Glu Phe Ser Asn Leu Tyr Arg Gln
260 265 270
Gln His Pro Glu Val Ala Ser Leu Arg Ile Lys Met Val Pro Leu Tyr
275 280 285
Lys Gln Ile Leu Ser Asp Arg Thr Thr Leu Ser Phe Val Pro Glu Ala
290 295 300
Leu Lys Asp Asp Glu Gln Ala Ile Asn Ala Val Asp Gly Leu Arg Ser
305 310 315 320
Glu Leu Glu Arg Asn Asp Ile Phe Asn Arg Ile Lys Arg Leu Phe Gly
325 330 335
Lys Asn Asn Leu Tyr Ser Leu Asp Lys Ile Trp Ile Lys Asn Ser Ser
340 345 350
Ile Ser Ala Phe Ser Asn Glu Leu Phe Lys Asn Trp Ser Phe Ile Glu
355 360 365
Asp Ala Leu Lys Glu Phe Lys Glu Asn Glu Phe Asn Gly Ala Arg Ser
370 375 380
Ala Gly Lys Lys Ala Glu Lys Trp Leu Lys Ser Lys Tyr Phe Ser Phe
385 390 395 400
Ala Asp Ile Asp Ala Ala Val Lys Ser Tyr Ser Glu Gln Val Ser Ala
405 410 415
Asp Ile Ser Ser Ala Pro Ser Ala Ser Tyr Phe Ala Lys Phe Thr Asn
420 425 430
Leu Ile Glu Thr Ala Ala Glu Asn Gly Arg Lys Phe Ser Tyr Phe Ala
435 440 445
Ala Glu Ser Lys Ala Phe Arg Gly Asp Asp Gly Lys Thr Glu Ile Ile
450 455 460
Lys Ala Tyr Leu Asp Ser Leu Asn Asp Ile Leu His Cys Leu Lys Pro
465 470 475 480
Phe Glu Thr Glu Asp Ile Ser Asp Ile Asp Thr Glu Phe Tyr Ser Ala
485 490 495
Phe Ala Glu Ile Tyr Asp Ser Val Lys Asp Val Ile Pro Val Tyr Asn
500 505 510
Ala Val Arg Asn Tyr Thr Thr Gln Lys Pro Phe Ser Thr Glu Lys Phe
515 520 525
Lys Leu Asn Phe Glu Asn Pro Ala Leu Ala Lys Gly Trp Asp Lys Asn
530 535 540
Lys Glu Gln Asn Asn Thr Ala Ile Ile Leu Met Lys Asp Gly Lys Tyr
545 550 555 560
Tyr Leu Gly Val Ile Asp Lys Asn Asn Lys Leu Arg Ala Asp Asp Leu
565 570 575
Ala Asp Asp Gly Ser Ala Tyr Gly Tyr Met Lys Met Asn Tyr Lys Phe
580 585 590
Ile Pro Thr Pro His Met Glu Leu Pro Lys Val Phe Leu Pro Lys Arg
595 600 605
Ala Pro Lys Arg Tyr Asn Pro Ser Arg Glu Ile Leu Leu Ile Lys Glu
610 615 620
Asn Lys Thr Phe Ile Lys Asp Lys Asn Phe Asn Arg Thr Asp Cys His
625 630 635 640
Lys Leu Ile Asp Phe Phe Lys Asp Ser Ile Asn Lys His Lys Asp Trp
645 650 655
Arg Thr Phe Gly Phe Asp Phe Ser Asp Thr Asp Ser Tyr Glu Asp Ile
660 665 670
Ser Asp Phe Tyr Met Glu Val Gln Asp Gln Gly Tyr Lys Leu Thr Phe
675 680 685
Thr Arg Leu Ser Ala Glu Lys Ile Asp Lys Trp Val Glu Glu Gly Arg
690 695 700
Leu Phe Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ala Asp Gly Ala Gln
705 710 715 720
Gly Ser Pro Asn Leu His Thr Leu Tyr Trp Lys Ala Ile Phe Ser Glu
725 730 735
Glu Asn Leu Lys Asp Val Val Leu Lys Leu Asn Gly Glu Ala Glu Leu
740 745 750
Phe Phe Arg Arg Lys Ser Ile Asp Lys Pro Ala Val His Ala Lys Gly
755 760 765
Ser Met Lys Val Asn Arg Arg Asp Ile Asp Gly Asn Pro Ile Asp Glu
770 775 780
Gly Thr Tyr Val Glu Ile Cys Gly Tyr Ala Asn Gly Lys Arg Asp Met
785 790 795 800
Ala Ser Leu Asn Ala Gly Ala Arg Gly Leu Ile Glu Ser Gly Leu Val
805 810 815
Arg Ile Thr Glu Val Lys His Glu Leu Val Lys Asp Lys Arg Tyr Thr
820 825 830
Ile Asp Lys Tyr Phe Phe His Val Pro Phe Thr Ile Asn Phe Lys Ala
835 840 845
Gln Gly Gln Gly Asn Ile Asn Ser Asp Val Asn Leu Phe Leu Arg Asn
850 855 860
Asn Lys Asp Val Asn Ile Ile Gly Ile Asp Arg Gly Glu Arg Asn Leu
865 870 875 880
Val Tyr Val Ser Leu Ile Asp Arg Asp Gly His Ile Lys Leu Gln Lys
885 890 895
Asp Phe Asn Ile Ile Gly Gly Met Asp Tyr His Ala Lys Leu Asn Gln
900 905 910
Lys Glu Lys Glu Arg Asp Thr Ala Arg Lys Ser Trp Lys Thr Ile Gly
915 920 925
Thr Ile Lys Glu Leu Lys Glu Gly Tyr Leu Ser Gln Val Val His Glu
930 935 940
Ile Val Arg Leu Ala Val Asp Asn Asn Ala Val Ile Val Met Glu Asp
945 950 955 960
Leu Asn Ile Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Val
965 970 975
Tyr Gln Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Leu Val
980 985 990
Phe Lys Asp Ala Gly Tyr Asp Ala Pro Cys Gly Ile Leu Lys Gly Leu
995 1000 1005
Gln Leu Thr Glu Lys Phe Glu Ser Phe Thr Lys Leu Gly Lys Gln
1010 1015 1020
Cys Gly Ile Ile Phe Tyr Ile Pro Ala Gly Tyr Thr Ser Lys Ile
1025 1030 1035
Asp Pro Thr Thr Gly Phe Val Asn Leu Phe Asn Ile Asn Asp Val
1040 1045 1050
Ser Ser Lys Glu Lys Gln Lys Asp Phe Ile Gly Lys Leu Asp Ser
1055 1060 1065
Ile Arg Phe Asp Ala Lys Arg Asp Met Phe Thr Phe Glu Phe Asp
1070 1075 1080
Tyr Asp Lys Phe Arg Thr Tyr Gln Thr Ser Tyr Arg Lys Lys Trp
1085 1090 1095
Ala Val Trp Thr Asn Gly Lys Arg Ile Val Arg Glu Lys Asp Lys
1100 1105 1110
Asp Gly Lys Phe Arg Met Asn Asp Arg Leu Leu Thr Glu Asp Met
1115 1120 1125
Lys Asn Ile Leu Asn Lys Tyr Ala Leu Ala Tyr Lys Ala Gly Glu
1130 1135 1140
Asp Ile Leu Pro Asp Val Ile Ser Arg Asp Lys Ser Leu Ala Ser
1145 1150 1155
Glu Ile Phe Tyr Val Phe Lys Asn Thr Leu Gln Met Arg Asn Ser
1160 1165 1170
Lys Arg Asp Thr Gly Glu Asp Phe Ile Ile Ser Pro Val Leu Asn
1175 1180 1185
Ala Lys Gly Arg Phe Phe Asp Ser Arg Lys Thr Asp Ala Ala Leu
1190 1195 1200
Pro Ile Asp Ala Asp Ala Asn Gly Ala Tyr His Ile Ala Leu Lys
1205 1210 1215
Gly Ser Leu Val Leu Asp Ala Ile Asp Glu Lys Leu Lys Glu Asp
1220 1225 1230
Gly Arg Ile Asp Tyr Lys Asp Met Ala Val Ser Asn Pro Lys Trp
1235 1240 1245
Phe Glu Phe Met Gln Thr Arg Lys Phe Asp Phe
1250 1255
<210> 56
<211> 1264
<212> PRT
<213> 短普雷沃氏菌
<220>
<221> MISC_FEATURE
<222> (1)..(1264)
<223> Genbank WP_044110123 Cpf1
<400> 56
Met Lys Gln Phe Thr Asn Leu Tyr Gln Leu Ser Lys Thr Leu Arg Phe
1 5 10 15
Glu Leu Lys Pro Ile Gly Lys Thr Leu Glu His Ile Asn Ala Asn Gly
20 25 30
Phe Ile Asp Asn Asp Ala His Arg Ala Glu Ser Tyr Lys Lys Val Lys
35 40 45
Lys Leu Ile Asp Asp Tyr His Lys Asp Tyr Ile Glu Asn Val Leu Asn
50 55 60
Asn Phe Lys Leu Asn Gly Glu Tyr Leu Gln Ala Tyr Phe Asp Leu Tyr
65 70 75 80
Ser Gln Asp Thr Lys Asp Lys Gln Phe Lys Asp Ile Gln Asp Lys Leu
85 90 95
Arg Lys Ser Ile Ala Ser Ala Leu Lys Gly Asp Asp Arg Tyr Lys Thr
100 105 110
Ile Asp Lys Lys Glu Leu Ile Arg Gln Asp Met Lys Thr Phe Leu Lys
115 120 125
Lys Asp Thr Asp Lys Ala Leu Leu Asp Glu Phe Tyr Glu Phe Thr Thr
130 135 140
Tyr Phe Thr Gly Tyr His Glu Asn Arg Lys Asn Met Tyr Ser Asp Glu
145 150 155 160
Ala Lys Ser Thr Ala Ile Ala Tyr Arg Leu Ile His Asp Asn Leu Pro
165 170 175
Lys Phe Ile Asp Asn Ile Ala Val Phe Lys Lys Ile Ala Asn Thr Ser
180 185 190
Val Ala Asp Asn Phe Ser Thr Ile Tyr Lys Asn Phe Glu Glu Tyr Leu
195 200 205
Asn Val Asn Ser Ile Asp Glu Ile Phe Ser Leu Asp Tyr Tyr Asn Ile
210 215 220
Val Leu Thr Gln Thr Gln Ile Glu Val Tyr Asn Ser Ile Ile Gly Gly
225 230 235 240
Arg Thr Leu Glu Asp Asp Thr Lys Ile Gln Gly Ile Asn Glu Phe Val
245 250 255
Asn Leu Tyr Asn Gln Gln Leu Ala Asn Lys Lys Asp Arg Leu Pro Lys
260 265 270
Leu Lys Pro Leu Phe Lys Gln Ile Leu Ser Asp Arg Val Gln Leu Ser
275 280 285
Trp Leu Gln Glu Glu Phe Asn Thr Gly Ala Asp Val Leu Asn Ala Val
290 295 300
Lys Glu Tyr Cys Thr Ser Tyr Phe Asp Asn Val Glu Glu Ser Val Lys
305 310 315 320
Val Leu Leu Thr Gly Ile Ser Asp Tyr Asp Leu Ser Lys Ile Tyr Ile
325 330 335
Thr Asn Asp Leu Ala Leu Thr Asp Val Ser Gln Arg Met Phe Gly Glu
340 345 350
Trp Ser Ile Ile Pro Asn Ala Ile Glu Gln Arg Leu Arg Ser Asp Asn
355 360 365
Pro Lys Lys Thr Asn Glu Lys Glu Glu Lys Tyr Ser Asp Arg Ile Ser
370 375 380
Lys Leu Lys Lys Leu Pro Lys Ser Tyr Ser Leu Gly Tyr Ile Asn Glu
385 390 395 400
Cys Ile Ser Glu Leu Asn Gly Ile Asp Ile Ala Asp Tyr Tyr Ala Thr
405 410 415
Leu Gly Ala Ile Asn Thr Glu Ser Lys Gln Glu Pro Ser Ile Pro Thr
420 425 430
Ser Ile Gln Val His Tyr Asn Ala Leu Lys Pro Ile Leu Asp Thr Asp
435 440 445
Tyr Pro Arg Glu Lys Asn Leu Ser Gln Asp Lys Leu Thr Val Met Gln
450 455 460
Leu Lys Asp Leu Leu Asp Asp Phe Lys Ala Leu Gln His Phe Ile Lys
465 470 475 480
Pro Leu Leu Gly Asn Gly Asp Glu Ala Glu Lys Asp Glu Lys Phe Tyr
485 490 495
Gly Glu Leu Met Gln Leu Trp Glu Val Ile Asp Ser Ile Thr Pro Leu
500 505 510
Tyr Asn Lys Val Arg Asn Tyr Cys Thr Arg Lys Pro Phe Ser Thr Glu
515 520 525
Lys Ile Lys Val Asn Phe Glu Asn Ala Gln Leu Leu Asp Gly Trp Asp
530 535 540
Glu Asn Lys Glu Ser Thr Asn Ala Ser Ile Ile Leu Arg Lys Asn Gly
545 550 555 560
Met Tyr Tyr Leu Gly Ile Met Lys Lys Glu Tyr Arg Asn Ile Leu Thr
565 570 575
Lys Pro Met Pro Ser Asp Gly Asp Cys Tyr Asp Lys Val Val Tyr Lys
580 585 590
Phe Phe Lys Asp Ile Thr Thr Met Val Pro Lys Cys Thr Thr Gln Met
595 600 605
Lys Ser Val Lys Glu His Phe Ser Asn Ser Asn Asp Asp Tyr Thr Leu
610 615 620
Phe Glu Lys Asp Lys Phe Ile Ala Pro Val Val Ile Thr Lys Glu Ile
625 630 635 640
Phe Asp Leu Asn Asn Val Leu Tyr Asn Gly Val Lys Lys Phe Gln Ile
645 650 655
Gly Tyr Leu Asn Asn Thr Gly Asp Ser Phe Gly Tyr Asn His Ala Val
660 665 670
Glu Ile Trp Lys Ser Phe Cys Leu Lys Phe Leu Lys Ala Tyr Lys Ser
675 680 685
Thr Ser Ile Tyr Asp Phe Ser Ser Ile Glu Lys Asn Ile Gly Cys Tyr
690 695 700
Asn Asp Leu Asn Ser Phe Tyr Gly Ala Val Asn Leu Leu Leu Tyr Asn
705 710 715 720
Leu Thr Tyr Arg Lys Val Ser Val Asp Tyr Ile His Gln Leu Val Asp
725 730 735
Glu Asp Lys Met Tyr Leu Phe Met Ile Tyr Asn Lys Asp Phe Ser Thr
740 745 750
Tyr Ser Lys Gly Thr Pro Asn Met His Thr Leu Tyr Trp Lys Met Leu
755 760 765
Phe Asp Glu Ser Asn Leu Asn Asp Val Val Tyr Lys Leu Asn Gly Gln
770 775 780
Ala Glu Val Phe Tyr Arg Lys Lys Ser Ile Thr Tyr Gln His Pro Thr
785 790 795 800
His Pro Ala Asn Lys Pro Ile Asp Asn Lys Asn Val Asn Asn Pro Lys
805 810 815
Lys Gln Ser Asn Phe Glu Tyr Asp Leu Ile Lys Asp Lys Arg Tyr Thr
820 825 830
Val Asp Lys Phe Met Phe His Val Pro Ile Thr Leu Asn Phe Lys Gly
835 840 845
Met Gly Asn Gly Asp Ile Asn Met Gln Val Arg Glu Tyr Ile Lys Thr
850 855 860
Thr Asp Asp Leu His Phe Ile Gly Ile Asp Arg Gly Glu Arg His Leu
865 870 875 880
Leu Tyr Ile Cys Val Ile Asn Gly Lys Gly Glu Ile Val Glu Gln Tyr
885 890 895
Ser Leu Asn Glu Ile Val Asn Asn Tyr Lys Gly Thr Glu Tyr Lys Thr
900 905 910
Asp Tyr His Thr Leu Leu Ser Glu Arg Asp Lys Lys Arg Lys Glu Glu
915 920 925
Arg Ser Ser Trp Gln Thr Ile Glu Gly Ile Lys Glu Leu Lys Ser Gly
930 935 940
Tyr Leu Ser Gln Val Ile His Lys Ile Thr Gln Leu Met Ile Lys Tyr
945 950 955 960
Asn Ala Ile Val Leu Leu Glu Asp Leu Asn Met Gly Phe Lys Arg Gly
965 970 975
Arg Gln Lys Val Glu Ser Ser Val Tyr Gln Gln Phe Glu Lys Ala Leu
980 985 990
Ile Asp Lys Leu Asn Tyr Leu Val Asp Lys Asn Lys Asp Ala Asn Glu
995 1000 1005
Ile Gly Gly Leu Leu His Ala Tyr Gln Leu Thr Asn Asp Pro Lys
1010 1015 1020
Leu Pro Asn Lys Asn Ser Lys Gln Ser Gly Phe Leu Phe Tyr Val
1025 1030 1035
Pro Ala Trp Asn Thr Ser Lys Ile Asp Pro Val Thr Gly Phe Val
1040 1045 1050
Asn Leu Leu Asp Thr Arg Tyr Glu Asn Val Ala Lys Ala Gln Ala
1055 1060 1065
Phe Phe Lys Lys Phe Asp Ser Ile Arg Tyr Asn Lys Glu Tyr Asp
1070 1075 1080
Arg Phe Glu Phe Lys Phe Asp Tyr Ser Asn Phe Thr Ala Lys Ala
1085 1090 1095
Glu Asp Thr Arg Thr Gln Trp Thr Leu Cys Thr Tyr Gly Thr Arg
1100 1105 1110
Ile Glu Thr Phe Arg Asn Ala Glu Lys Asn Ser Asn Trp Asp Ser
1115 1120 1125
Arg Glu Ile Asp Leu Thr Thr Glu Trp Lys Thr Leu Phe Thr Gln
1130 1135 1140
His Asn Ile Pro Leu Asn Ala Asn Leu Lys Glu Ala Ile Leu Leu
1145 1150 1155
Gln Ala Asn Lys Asn Phe Tyr Thr Asp Ile Leu His Leu Met Lys
1160 1165 1170
Leu Thr Leu Gln Met Arg Asn Ser Val Thr Gly Thr Asp Ile Asp
1175 1180 1185
Tyr Met Val Ser Pro Val Ala Asn Glu Cys Gly Glu Phe Phe Asp
1190 1195 1200
Ser Arg Lys Val Lys Glu Gly Leu Pro Val Asn Ala Asp Ala Asn
1205 1210 1215
Gly Ala Tyr Asn Ile Ala Arg Lys Gly Leu Trp Leu Ala Gln Gln
1220 1225 1230
Ile Lys Asn Ala Asn Asp Leu Ser Asp Val Lys Leu Ala Ile Thr
1235 1240 1245
Asn Lys Glu Trp Leu Gln Phe Ala Gln Lys Lys Gln Tyr Leu Lys
1250 1255 1260
Asp
<210> 57
<211> 1273
<212> PRT
<213> 黄杆菌
<220>
<221> MISC_FEATURE
<222> (1)..(1273)
<223> Genbank WP_045971446 Cpf1
<400> 57
Met Lys Asn Phe Ser Asn Leu Tyr Gln Val Ser Lys Thr Val Arg Phe
1 5 10 15
Glu Leu Lys Pro Ile Gly Asn Thr Leu Glu Asn Ile Lys Asn Lys Ser
20 25 30
Leu Leu Lys Asn Asp Ser Ile Arg Ala Glu Ser Tyr Gln Lys Met Lys
35 40 45
Lys Thr Ile Asp Glu Phe His Lys Tyr Phe Ile Asp Leu Ala Leu Asn
50 55 60
Asn Lys Lys Leu Ser Tyr Leu Asn Glu Tyr Ile Ala Leu Tyr Thr Gln
65 70 75 80
Ser Ala Glu Ala Lys Lys Glu Asp Lys Phe Lys Ala Asp Phe Lys Lys
85 90 95
Val Gln Asp Asn Leu Arg Lys Glu Ile Val Ser Ser Phe Thr Glu Gly
100 105 110
Glu Ala Lys Ala Ile Phe Ser Val Leu Asp Lys Lys Glu Leu Ile Thr
115 120 125
Ile Glu Leu Glu Lys Trp Lys Asn Glu Asn Asn Leu Ala Val Tyr Leu
130 135 140
Asp Glu Ser Phe Lys Ser Phe Thr Thr Tyr Phe Thr Gly Phe His Gln
145 150 155 160
Asn Arg Lys Asn Met Tyr Ser Ala Glu Ala Asn Ser Thr Ala Ile Ala
165 170 175
Tyr Arg Leu Ile His Glu Asn Leu Pro Lys Phe Ile Glu Asn Ser Lys
180 185 190
Ala Phe Glu Lys Ser Ser Gln Ile Ala Glu Leu Gln Pro Lys Ile Glu
195 200 205
Lys Leu Tyr Lys Glu Phe Glu Ala Tyr Leu Asn Val Asn Ser Ile Ser
210 215 220
Glu Leu Phe Glu Ile Asp Tyr Phe Asn Glu Val Leu Thr Gln Lys Gly
225 230 235 240
Ile Thr Val Tyr Asn Asn Ile Ile Gly Gly Arg Thr Ala Thr Glu Gly
245 250 255
Lys Gln Lys Ile Gln Gly Leu Asn Glu Ile Ile Asn Leu Tyr Asn Gln
260 265 270
Thr Lys Pro Lys Asn Glu Arg Leu Pro Lys Leu Lys Gln Leu Tyr Lys
275 280 285
Gln Ile Leu Ser Asp Arg Ile Ser Leu Ser Phe Leu Pro Asp Ala Phe
290 295 300
Thr Glu Gly Lys Gln Val Leu Lys Ala Val Phe Glu Phe Tyr Lys Ile
305 310 315 320
Asn Leu Leu Ser Tyr Lys Gln Asp Gly Val Glu Glu Ser Gln Asn Leu
325 330 335
Leu Glu Leu Ile Gln Gln Val Val Lys Asn Leu Gly Asn Gln Asp Val
340 345 350
Asn Lys Ile Tyr Leu Lys Asn Asp Thr Ser Leu Thr Thr Ile Ala Gln
355 360 365
Gln Leu Phe Gly Asp Phe Ser Val Phe Ser Ala Ala Leu Gln Tyr Arg
370 375 380
Tyr Glu Thr Val Val Asn Pro Lys Tyr Thr Ala Glu Tyr Gln Lys Ala
385 390 395 400
Asn Glu Ala Lys Gln Glu Lys Leu Asp Lys Glu Lys Ile Lys Phe Val
405 410 415
Lys Gln Asp Tyr Phe Ser Ile Ala Phe Leu Gln Glu Val Val Ala Asp
420 425 430
Tyr Val Lys Thr Leu Asp Glu Asn Leu Asp Trp Lys Gln Lys Tyr Thr
435 440 445
Pro Ser Cys Ile Ala Asp Tyr Phe Thr Thr His Phe Ile Ala Lys Lys
450 455 460
Glu Asn Glu Ala Asp Lys Thr Phe Asn Phe Ile Ala Asn Ile Lys Ala
465 470 475 480
Lys Tyr Gln Cys Ile Gln Gly Ile Leu Glu Gln Ala Asp Asp Tyr Glu
485 490 495
Asp Glu Leu Lys Gln Asp Gln Lys Leu Ile Asp Asn Ile Lys Phe Phe
500 505 510
Leu Asp Ala Ile Leu Glu Val Val His Phe Ile Lys Pro Leu His Leu
515 520 525
Lys Ser Glu Ser Ile Thr Glu Lys Asp Asn Ala Phe Tyr Asp Val Phe
530 535 540
Glu Asn Tyr Tyr Glu Ala Leu Asn Val Val Thr Pro Leu Tyr Asn Met
545 550 555 560
Val Arg Asn Tyr Val Thr Gln Lys Pro Tyr Ser Thr Glu Lys Ile Lys
565 570 575
Leu Asn Phe Glu Asn Ala Gln Leu Leu Asn Gly Trp Asp Ala Asn Lys
580 585 590
Glu Lys Asp Tyr Leu Thr Thr Ile Leu Lys Arg Asp Gly Asn Tyr Phe
595 600 605
Leu Ala Ile Met Asp Lys Lys His Asn Lys Thr Phe Gln Gln Phe Thr
610 615 620
Glu Asp Asp Glu Asn Tyr Glu Lys Ile Val Tyr Lys Leu Leu Pro Gly
625 630 635 640
Val Asn Lys Met Leu Pro Lys Val Phe Phe Ser Asn Lys Asn Ile Ala
645 650 655
Phe Phe Asn Pro Ser Lys Glu Ile Leu Asp Asn Tyr Lys Asn Asn Thr
660 665 670
His Lys Lys Gly Ala Thr Phe Asn Leu Lys Asp Cys His Ala Leu Ile
675 680 685
Asp Phe Phe Lys Asp Ser Leu Asn Lys His Glu Asp Trp Lys Tyr Phe
690 695 700
Asp Phe Gln Phe Ser Glu Thr Lys Thr Tyr Gln Asp Leu Ser Gly Phe
705 710 715 720
Tyr Lys Glu Val Glu His Gln Gly Tyr Lys Ile Asn Phe Lys Lys Val
725 730 735
Ser Val Ser Gln Ile Asp Thr Leu Ile Glu Glu Gly Lys Met Tyr Leu
740 745 750
Phe Gln Ile Tyr Asn Lys Asp Phe Ser Pro Tyr Ala Lys Gly Lys Pro
755 760 765
Asn Met His Thr Leu Tyr Trp Lys Ala Leu Phe Glu Thr Gln Asn Leu
770 775 780
Glu Asn Val Ile Tyr Lys Leu Asn Gly Gln Ala Glu Ile Phe Phe Arg
785 790 795 800
Lys Ala Ser Ile Lys Lys Lys Asn Ile Ile Thr His Lys Ala His Gln
805 810 815
Pro Ile Ala Ala Lys Asn Pro Leu Thr Pro Thr Ala Lys Asn Thr Phe
820 825 830
Ala Tyr Asp Leu Ile Lys Asp Lys Arg Tyr Thr Val Asp Lys Phe Gln
835 840 845
Phe His Val Pro Ile Thr Met Asn Phe Lys Ala Thr Gly Asn Ser Tyr
850 855 860
Ile Asn Gln Asp Val Leu Ala Tyr Leu Lys Asp Asn Pro Glu Val Asn
865 870 875 880
Ile Ile Gly Leu Asp Arg Gly Glu Arg His Leu Val Tyr Leu Thr Leu
885 890 895
Ile Asp Gln Lys Gly Thr Ile Leu Leu Gln Glu Ser Leu Asn Val Ile
900 905 910
Gln Asp Glu Lys Thr His Thr Pro Tyr His Thr Leu Leu Asp Asn Lys
915 920 925
Glu Ile Ala Arg Asp Lys Ala Arg Lys Asn Trp Gly Ser Ile Glu Ser
930 935 940
Ile Lys Glu Leu Lys Glu Gly Tyr Ile Ser Gln Val Val His Lys Ile
945 950 955 960
Thr Lys Met Met Ile Glu His Asn Ala Ile Val Val Met Glu Asp Leu
965 970 975
Asn Phe Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Ile Tyr
980 985 990
Gln Lys Leu Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Leu Val Leu
995 1000 1005
Lys Asp Lys Gln Pro His Glu Leu Gly Gly Leu Tyr Asn Ala Leu
1010 1015 1020
Gln Leu Thr Asn Lys Phe Glu Ser Phe Gln Lys Met Gly Lys Gln
1025 1030 1035
Ser Gly Phe Leu Phe Tyr Val Pro Ala Trp Asn Thr Ser Lys Ile
1040 1045 1050
Asp Pro Thr Thr Gly Phe Val Asn Tyr Phe Tyr Thr Lys Tyr Glu
1055 1060 1065
Asn Val Glu Lys Ala Lys Thr Phe Phe Ser Lys Phe Asp Ser Ile
1070 1075 1080
Leu Tyr Asn Lys Thr Lys Gly Tyr Phe Glu Phe Val Val Lys Asn
1085 1090 1095
Tyr Ser Asp Phe Asn Pro Lys Ala Ala Asp Thr Arg Gln Glu Trp
1100 1105 1110
Thr Ile Cys Thr His Gly Glu Arg Ile Glu Thr Lys Arg Gln Lys
1115 1120 1125
Glu Gln Asn Asn Asn Phe Val Ser Thr Thr Ile Gln Leu Thr Glu
1130 1135 1140
Gln Phe Val Asn Phe Phe Glu Lys Val Gly Leu Asp Leu Ser Lys
1145 1150 1155
Glu Leu Lys Thr Gln Leu Ile Ala Gln Asn Glu Lys Ser Phe Phe
1160 1165 1170
Glu Glu Leu Phe His Leu Leu Lys Leu Thr Leu Gln Met Arg Asn
1175 1180 1185
Ser Glu Ser His Thr Glu Ile Asp Tyr Leu Ile Ser Pro Val Ala
1190 1195 1200
Asn Glu Lys Gly Ile Phe Tyr Asp Ser Arg Lys Ala Thr Ala Ser
1205 1210 1215
Leu Pro Ile Asp Ala Asp Ala Asn Gly Ala Tyr His Ile Ala Lys
1220 1225 1230
Lys Gly Leu Trp Ile Met Glu Gln Ile Asn Lys Thr Asn Ser Glu
1235 1240 1245
Asp Asp Leu Lys Lys Val Lys Leu Ala Ile Ser Asn Arg Glu Trp
1250 1255 1260
Leu Gln Tyr Val Gln Gln Val Gln Lys Lys
1265 1270
<210> 58
<211> 973
<212> PRT
<213> 羊膜斯聂氏菌(Sneathia amnii)
<220>
<221> MISC_FEATURE
<222> (1)..(973)
<223> Genbank WP_046328599 Cpf1
<400> 58
Met Thr Glu Glu Asp Thr Lys Ser Phe Val Asp Glu Ile Leu Leu Thr
1 5 10 15
Pro Glu Ser Val Ile Lys Thr Ile Asp Asn Phe Ile Asp Ser Ile Ile
20 25 30
Met Asn Asp Ile Glu Gly Leu Lys Glu Glu Phe Leu Lys Ile Ser Leu
35 40 45
Glu Asn Phe Glu Gly Ile Tyr Ile Ser Asn Lys Lys Leu Asn Glu Ile
50 55 60
Ser Asn Arg Lys Phe Gly Asp Tyr Asn Ser Ile Asn Met Met Ile Lys
65 70 75 80
Gln Ser Met Asn Glu Lys Gly Ile Leu Ser Lys Lys Glu Ile Asn Glu
85 90 95
Leu Ile Pro Asp Leu Glu Asn Ile Asn Lys Pro Lys Val Lys Ser Phe
100 105 110
Asn Leu Ser Phe Ile Phe Glu Asn Leu Thr Lys Glu His Lys Glu Leu
115 120 125
Ile Ile Asp Tyr Ile Arg Glu Asn Ile Cys Asn Val Ile Glu Asn Val
130 135 140
Lys Ile Thr Ile Glu Lys Tyr Arg Asn Ile Asp Asn Lys Ile Glu Phe
145 150 155 160
Lys Asn Asn Ala Glu Lys Val Ser Lys Ile Lys Glu Met Leu Glu Ser
165 170 175
Ile Asn Glu Leu Cys Lys Leu Ile Lys Glu Phe Asn Thr Asp Glu Ile
180 185 190
Glu Lys Asn Asn Glu Phe Tyr Asn Ile Leu Asn Lys Asn Phe Glu Ile
195 200 205
Phe Glu Ser Ser Tyr Lys Val Leu Asn Lys Val Arg Asn Phe Val Thr
210 215 220
Lys Lys Glu Val Ile Glu Asn Lys Met Lys Leu Asn Phe Ser Asn Tyr
225 230 235 240
Gln Leu Gly Asn Gly Trp His Lys Asn Lys Glu Lys Asp Cys Ser Ile
245 250 255
Ile Leu Phe Arg Lys Arg Asn Asn Glu Arg Trp Ile Tyr Tyr Leu Gly
260 265 270
Ile Leu Lys His Gly Thr Lys Ile Lys Glu Asn Asp Tyr Leu Ser Ser
275 280 285
Val Asp Thr Gly Phe Tyr Lys Met Asp Tyr Tyr Ala Gln Asn Ser Leu
290 295 300
Ser Lys Met Ile Pro Lys Cys Ser Ile Thr Val Lys Asn Val Lys Asn
305 310 315 320
Ala Pro Glu Asp Glu Ser Val Ile Leu Asn Asp Ser Lys Lys Phe Asn
325 330 335
Glu Pro Leu Glu Ile Thr Pro Glu Ile Arg Lys Leu Tyr Gly Asn Asn
340 345 350
Glu His Ile Lys Gly Asp Lys Phe Lys Lys Glu Ser Leu Val Lys Trp
355 360 365
Ile Asp Phe Cys Lys Glu Phe Leu Leu Lys Tyr Lys Ser Phe Glu Lys
370 375 380
Ala Lys Lys Glu Ile Leu Lys Leu Lys Glu Ser Asn Leu Tyr Glu Asn
385 390 395 400
Leu Glu Glu Phe Tyr Ser Asp Ala Glu Glu Lys Ala Tyr Phe Leu Glu
405 410 415
Phe Ile Asn Ile Asp Glu Asp Lys Ile Lys Lys Leu Val Lys Glu Lys
420 425 430
Asn Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Ala Tyr Ser
435 440 445
Thr Gly Asn Lys Asn Leu His Thr Met Tyr Phe Glu Glu Leu Phe Thr
450 455 460
Asp Glu Asn Leu Lys Lys Pro Val Phe Lys Leu Asn Gly Asn Thr Glu
465 470 475 480
Val Phe Tyr Arg Ile Ala Ser Ser Lys Pro Lys Ile Val His Asn Lys
485 490 495
Gly Glu Lys Leu Val Asn Lys Thr Tyr Leu Asp Asp Gly Ile Ile Lys
500 505 510
Thr Ile Pro Asp Ser Val Tyr Glu Glu Ile Ser Glu Lys Val Lys Asn
515 520 525
Asn Glu Asp Tyr Ser Lys Leu Leu Glu Glu Asn Asn Ile Lys Asn Leu
530 535 540
Glu Ile Lys Val Ala Thr His Glu Ile Val Lys Asp Lys Arg Tyr Phe
545 550 555 560
Glu Asn Lys Phe Leu Phe Tyr Leu Pro Ile Thr Leu Asn Lys Lys Val
565 570 575
Ser Asn Lys Asn Thr Asn Lys Asn Ile Asn Lys Asn Val Ile Asp Glu
580 585 590
Ile Lys Asp Cys Asn Glu Tyr Asn Val Ile Gly Ile Asp Arg Gly Glu
595 600 605
Arg Asn Leu Ile Ser Leu Cys Ile Ile Asn Gln Asn Gly Glu Ile Ile
610 615 620
Leu Gln Lys Glu Met Asn Ile Ile Gln Ser Ser Asp Lys Tyr Asn Val
625 630 635 640
Asp Tyr Asn Glu Lys Leu Glu Ile Lys Ser Lys Glu Arg Asp Asn Ala
645 650 655
Lys Lys Asn Trp Ser Glu Ile Gly Lys Ile Lys Asp Leu Lys Ser Gly
660 665 670
Tyr Leu Ser Ala Val Val His Glu Ile Val Lys Leu Ala Ile Glu Tyr
675 680 685
Asn Ala Val Ile Ile Leu Glu Asp Leu Asn Asn Gly Phe Lys Asn Ser
690 695 700
Arg Lys Lys Val Asp Lys Gln Ile Tyr Gln Lys Phe Glu Arg Ala Leu
705 710 715 720
Ile Glu Lys Leu Gln Phe Leu Ile Phe Lys Asn Tyr Asp Lys Asn Glu
725 730 735
Lys Gly Gly Leu Arg Asn Ala Phe Gln Leu Thr Pro Glu Leu Lys Asn
740 745 750
Ile Thr Lys Val Ala Ser Gln Gln Gly Ile Ile Ile Tyr Thr Asn Pro
755 760 765
Ala Tyr Thr Ser Lys Ile Asp Pro Thr Thr Gly Tyr Ala Asn Ile Ile
770 775 780
Lys Lys Ser Asn Asn Asn Glu Glu Ser Ile Val Lys Ala Ile Asp Lys
785 790 795 800
Ile Ser Tyr Asp Lys Glu Lys Asp Met Phe Tyr Phe Asp Ile Asn Leu
805 810 815
Ser Asn Ser Ser Phe Asn Leu Thr Val Lys Asn Val Leu Lys Lys Glu
820 825 830
Trp Arg Ile Tyr Thr Asn Gly Glu Arg Ile Ile Tyr Lys Asp Arg Lys
835 840 845
Tyr Ile Thr Leu Asn Ile Thr Gln Glu Met Lys Asp Ile Leu Ser Lys
850 855 860
Cys Gly Ile Asp Tyr Leu Asn Ile Asp Asn Leu Lys Gln Asp Ile Leu
865 870 875 880
Lys Asn Lys Leu His Lys Lys Val Tyr Tyr Ile Phe Glu Leu Ala Asn
885 890 895
Lys Met Arg Asn Glu Asn Lys Asp Val Asp Tyr Ile Ile Ser Pro Val
900 905 910
Leu Asn Lys Asp Gly Lys Phe Phe Met Thr Gln Glu Ile Asn Glu Leu
915 920 925
Thr Pro Lys Asp Ala Asp Leu Asn Gly Ala Tyr Asn Ile Ala Leu Lys
930 935 940
Gly Lys Leu Met Ile Asp Asn Leu Asn Lys Lys Glu Lys Phe Val Phe
945 950 955 960
Leu Ser Asn Glu Asp Trp Leu Asn Phe Ile Gln Gly Arg
965 970
<210> 59
<211> 1238
<212> PRT
<213> 产甲烷菌暂定种
<220>
<221> MISC_FEATURE
<222> (1)..(1238)
<223> Genbank WP_048112740 Cpf1
<400> 59
Met Asn Asn Tyr Asp Glu Phe Thr Lys Leu Tyr Pro Ile Gln Lys Thr
1 5 10 15
Ile Arg Phe Glu Leu Lys Pro Gln Gly Arg Thr Met Glu His Leu Glu
20 25 30
Thr Phe Asn Phe Phe Glu Glu Asp Arg Asp Arg Ala Glu Lys Tyr Lys
35 40 45
Ile Leu Lys Glu Ala Ile Asp Glu Tyr His Lys Lys Phe Ile Asp Glu
50 55 60
His Leu Thr Asn Met Ser Leu Asp Trp Asn Ser Leu Lys Gln Ile Ser
65 70 75 80
Glu Lys Tyr Tyr Lys Ser Arg Glu Glu Lys Asp Lys Lys Val Phe Leu
85 90 95
Ser Glu Gln Lys Arg Met Arg Gln Glu Ile Val Ser Glu Phe Lys Lys
100 105 110
Asp Asp Arg Phe Lys Asp Leu Phe Ser Lys Lys Leu Phe Ser Glu Leu
115 120 125
Leu Lys Glu Glu Ile Tyr Lys Lys Gly Asn His Gln Glu Ile Asp Ala
130 135 140
Leu Lys Ser Phe Asp Lys Phe Ser Gly Tyr Phe Ile Gly Leu His Glu
145 150 155 160
Asn Arg Lys Asn Met Tyr Ser Asp Gly Asp Glu Ile Thr Ala Ile Ser
165 170 175
Asn Arg Ile Val Asn Glu Asn Phe Pro Lys Phe Leu Asp Asn Leu Gln
180 185 190
Lys Tyr Gln Glu Ala Arg Lys Lys Tyr Pro Glu Trp Ile Ile Lys Ala
195 200 205
Glu Ser Ala Leu Val Ala His Asn Ile Lys Met Asp Glu Val Phe Ser
210 215 220
Leu Glu Tyr Phe Asn Lys Val Leu Asn Gln Glu Gly Ile Gln Arg Tyr
225 230 235 240
Asn Leu Ala Leu Gly Gly Tyr Val Thr Lys Ser Gly Glu Lys Met Met
245 250 255
Gly Leu Asn Asp Ala Leu Asn Leu Ala His Gln Ser Glu Lys Ser Ser
260 265 270
Lys Gly Arg Ile His Met Thr Pro Leu Phe Lys Gln Ile Leu Ser Glu
275 280 285
Lys Glu Ser Phe Ser Tyr Ile Pro Asp Val Phe Thr Glu Asp Ser Gln
290 295 300
Leu Leu Pro Ser Ile Gly Gly Phe Phe Ala Gln Ile Glu Asn Asp Lys
305 310 315 320
Asp Gly Asn Ile Phe Asp Arg Ala Leu Glu Leu Ile Ser Ser Tyr Ala
325 330 335
Glu Tyr Asp Thr Glu Arg Ile Tyr Ile Arg Gln Ala Asp Ile Asn Arg
340 345 350
Val Ser Asn Val Ile Phe Gly Glu Trp Gly Thr Leu Gly Gly Leu Met
355 360 365
Arg Glu Tyr Lys Ala Asp Ser Ile Asn Asp Ile Asn Leu Glu Arg Thr
370 375 380
Cys Lys Lys Val Asp Lys Trp Leu Asp Ser Lys Glu Phe Ala Leu Ser
385 390 395 400
Asp Val Leu Glu Ala Ile Lys Arg Thr Gly Asn Asn Asp Ala Phe Asn
405 410 415
Glu Tyr Ile Ser Lys Met Arg Thr Ala Arg Glu Lys Ile Asp Ala Ala
420 425 430
Arg Lys Glu Met Lys Phe Ile Ser Glu Lys Ile Ser Gly Asp Glu Glu
435 440 445
Ser Ile His Ile Ile Lys Thr Leu Leu Asp Ser Val Gln Gln Phe Leu
450 455 460
His Phe Phe Asn Leu Phe Lys Ala Arg Gln Asp Ile Pro Leu Asp Gly
465 470 475 480
Ala Phe Tyr Ala Glu Phe Asp Glu Val His Ser Lys Leu Phe Ala Ile
485 490 495
Val Pro Leu Tyr Asn Lys Val Arg Asn Tyr Leu Thr Lys Asn Asn Leu
500 505 510
Asn Thr Lys Lys Ile Lys Leu Asn Phe Lys Asn Pro Thr Leu Ala Asn
515 520 525
Gly Trp Asp Gln Asn Lys Val Tyr Asp Tyr Ala Ser Leu Ile Phe Leu
530 535 540
Arg Asp Gly Asn Tyr Tyr Leu Gly Ile Ile Asn Pro Lys Arg Lys Lys
545 550 555 560
Asn Ile Lys Phe Glu Gln Gly Ser Gly Asn Gly Pro Phe Tyr Arg Lys
565 570 575
Met Val Tyr Lys Gln Ile Pro Gly Pro Asn Lys Asn Leu Pro Arg Val
580 585 590
Phe Leu Thr Ser Thr Lys Gly Lys Lys Glu Tyr Lys Pro Ser Lys Glu
595 600 605
Ile Ile Glu Gly Tyr Glu Ala Asp Lys His Ile Arg Gly Asp Lys Phe
610 615 620
Asp Leu Asp Phe Cys His Lys Leu Ile Asp Phe Phe Lys Glu Ser Ile
625 630 635 640
Glu Lys His Lys Asp Trp Ser Lys Phe Asn Phe Tyr Phe Ser Pro Thr
645 650 655
Glu Ser Tyr Gly Asp Ile Ser Glu Phe Tyr Leu Asp Val Glu Lys Gln
660 665 670
Gly Tyr Arg Met His Phe Glu Asn Ile Ser Ala Glu Thr Ile Asp Glu
675 680 685
Tyr Val Glu Lys Gly Asp Leu Phe Leu Phe Gln Ile Tyr Asn Lys Asp
690 695 700
Phe Val Lys Ala Ala Thr Gly Lys Lys Asp Met His Thr Ile Tyr Trp
705 710 715 720
Asn Ala Ala Phe Ser Pro Glu Asn Leu Gln Asp Val Val Val Lys Leu
725 730 735
Asn Gly Glu Ala Glu Leu Phe Tyr Arg Asp Lys Ser Asp Ile Lys Glu
740 745 750
Ile Val His Arg Glu Gly Glu Ile Leu Val Asn Arg Thr Tyr Asn Gly
755 760 765
Arg Thr Pro Val Pro Asp Lys Ile His Lys Lys Leu Thr Asp Tyr His
770 775 780
Asn Gly Arg Thr Lys Asp Leu Gly Glu Ala Lys Glu Tyr Leu Asp Lys
785 790 795 800
Val Arg Tyr Phe Lys Ala His Tyr Asp Ile Thr Lys Asp Arg Arg Tyr
805 810 815
Leu Asn Asp Lys Ile Tyr Phe His Val Pro Leu Thr Leu Asn Phe Lys
820 825 830
Ala Asn Gly Lys Lys Asn Leu Asn Lys Met Val Ile Glu Lys Phe Leu
835 840 845
Ser Asp Glu Lys Ala His Ile Ile Gly Ile Asp Arg Gly Glu Arg Asn
850 855 860
Leu Leu Tyr Tyr Ser Ile Ile Asp Arg Ser Gly Lys Ile Ile Asp Gln
865 870 875 880
Gln Ser Leu Asn Val Ile Asp Gly Phe Asp Tyr Arg Glu Lys Leu Asn
885 890 895
Gln Arg Glu Ile Glu Met Lys Asp Ala Arg Gln Ser Trp Asn Ala Ile
900 905 910
Gly Lys Ile Lys Asp Leu Lys Glu Gly Tyr Leu Ser Lys Ala Val His
915 920 925
Glu Ile Thr Lys Met Ala Ile Gln Tyr Asn Ala Ile Val Val Met Glu
930 935 940
Glu Leu Asn Tyr Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln
945 950 955 960
Ile Tyr Gln Lys Phe Glu Asn Met Leu Ile Asp Lys Met Asn Tyr Leu
965 970 975
Val Phe Lys Asp Ala Pro Asp Glu Ser Pro Gly Gly Val Leu Asn Ala
980 985 990
Tyr Gln Leu Thr Asn Pro Leu Glu Ser Phe Ala Lys Leu Gly Lys Gln
995 1000 1005
Thr Gly Ile Leu Phe Tyr Val Pro Ala Ala Tyr Thr Ser Lys Ile
1010 1015 1020
Asp Pro Thr Thr Gly Phe Val Asn Leu Phe Asn Thr Ser Ser Lys
1025 1030 1035
Thr Asn Ala Gln Glu Arg Lys Glu Phe Leu Gln Lys Phe Glu Ser
1040 1045 1050
Ile Ser Tyr Ser Ala Lys Asp Gly Gly Ile Phe Ala Phe Ala Phe
1055 1060 1065
Asp Tyr Arg Lys Phe Gly Thr Ser Lys Thr Asp His Lys Asn Val
1070 1075 1080
Trp Thr Ala Tyr Thr Asn Gly Glu Arg Met Arg Tyr Ile Lys Glu
1085 1090 1095
Lys Lys Arg Asn Glu Leu Phe Asp Pro Ser Lys Glu Ile Lys Glu
1100 1105 1110
Ala Leu Thr Ser Ser Gly Ile Lys Tyr Asp Gly Gly Gln Asn Ile
1115 1120 1125
Leu Pro Asp Ile Leu Arg Ser Asn Asn Asn Gly Leu Ile Tyr Thr
1130 1135 1140
Met Tyr Ser Ser Phe Ile Ala Ala Ile Gln Met Arg Val Tyr Asp
1145 1150 1155
Gly Lys Glu Asp Tyr Ile Ile Ser Pro Ile Lys Asn Ser Lys Gly
1160 1165 1170
Glu Phe Phe Arg Thr Asp Pro Lys Arg Arg Glu Leu Pro Ile Asp
1175 1180 1185
Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala Leu Arg Gly Glu Leu
1190 1195 1200
Thr Met Arg Ala Ile Ala Glu Lys Phe Asp Pro Asp Ser Glu Lys
1205 1210 1215
Met Ala Lys Leu Glu Leu Lys His Lys Asp Trp Phe Glu Phe Met
1220 1225 1230
Gln Thr Arg Gly Asp
1235
<210> 60
<211> 1222
<212> PRT
<213> 口腔杆菌(Oribacterium sp.)
<220>
<221> MISC_FEATURE
<222> (1)..(1222)
<223> Genbank WP_049895985 Cpf1
<400> 60
Met Glu Thr Glu Ile Leu Lys Tyr Asp Phe Phe Glu Arg Glu Gly Lys
1 5 10 15
Tyr Met Tyr Tyr Asp Gly Leu Thr Lys Gln Tyr Ala Leu Ser Lys Thr
20 25 30
Ile Arg Asn Glu Leu Val Pro Ile Gly Lys Thr Leu Asp Asn Ile Lys
35 40 45
Lys Asn Arg Ile Leu Glu Ala Asp Ile Lys Arg Lys Ser Asp Tyr Glu
50 55 60
His Val Lys Lys Leu Met Asp Met Tyr His Lys Lys Ile Ile Asn Glu
65 70 75 80
Ala Leu Asp Asn Phe Lys Leu Ser Val Leu Glu Asp Ala Ala Asp Ile
85 90 95
Tyr Phe Asn Lys Gln Asn Asp Glu Arg Asp Ile Asp Ala Phe Leu Lys
100 105 110
Ile Gln Asp Lys Leu Arg Lys Glu Ile Val Glu Gln Leu Lys Gly His
115 120 125
Thr Asp Tyr Ser Lys Val Gly Asn Lys Asp Phe Leu Gly Leu Leu Lys
130 135 140
Ala Ala Ser Thr Glu Glu Asp Arg Ile Leu Ile Glu Ser Phe Asp Asn
145 150 155 160
Phe Tyr Thr Tyr Phe Thr Ser Tyr Asn Lys Val Arg Ser Asn Leu Tyr
165 170 175
Ser Ala Glu Asp Lys Ser Ser Thr Val Ala Tyr Arg Leu Ile Asn Glu
180 185 190
Asn Leu Pro Lys Phe Phe Asp Asn Ile Lys Ala Tyr Arg Thr Val Arg
195 200 205
Asn Ala Gly Val Ile Ser Gly Asp Met Ser Ile Val Glu Gln Asp Glu
210 215 220
Leu Phe Glu Val Asp Thr Phe Asn His Thr Leu Thr Gln Tyr Gly Ile
225 230 235 240
Asp Thr Tyr Asn His Met Ile Gly Gln Leu Asn Ser Ala Ile Asn Leu
245 250 255
Tyr Asn Gln Lys Met His Gly Ala Gly Ser Phe Lys Lys Leu Pro Lys
260 265 270
Met Lys Glu Leu Tyr Lys Gln Leu Leu Thr Glu Arg Glu Glu Glu Phe
275 280 285
Ile Glu Glu Tyr Thr Asp Asp Glu Val Leu Ile Thr Ser Val His Asn
290 295 300
Tyr Val Ser Tyr Leu Ile Asp Tyr Leu Asn Ser Asp Lys Val Glu Ser
305 310 315 320
Phe Phe Asp Thr Leu Arg Lys Ser Asp Gly Lys Glu Val Phe Ile Lys
325 330 335
Asn Asp Val Ser Lys Thr Thr Met Ser Asn Ile Leu Phe Asp Asn Trp
340 345 350
Ser Thr Ile Asp Asp Leu Ile Asn His Glu Tyr Asp Ser Ala Pro Glu
355 360 365
Asn Val Lys Lys Thr Lys Asp Asp Lys Tyr Phe Glu Lys Arg Gln Lys
370 375 380
Asp Leu Lys Lys Asn Lys Ser Tyr Ser Leu Ser Lys Ile Ala Ala Leu
385 390 395 400
Cys Arg Asp Thr Thr Ile Leu Glu Lys Tyr Ile Arg Arg Leu Val Asp
405 410 415
Asp Ile Glu Lys Ile Tyr Thr Ser Asn Asn Val Phe Ser Asp Ile Val
420 425 430
Leu Ser Lys His Asp Arg Ser Lys Lys Leu Ser Lys Asn Thr Asn Ala
435 440 445
Val Gln Ala Ile Lys Asn Met Leu Asp Ser Ile Lys Asp Phe Glu His
450 455 460
Asp Val Met Leu Ile Asn Gly Ser Gly Gln Glu Ile Lys Lys Asn Leu
465 470 475 480
Asn Val Tyr Ser Glu Gln Glu Ala Leu Ala Gly Ile Leu Arg Gln Val
485 490 495
Asp His Ile Tyr Asn Leu Thr Arg Asn Tyr Leu Thr Lys Lys Pro Phe
500 505 510
Ser Thr Glu Lys Ile Lys Leu Asn Phe Asn Arg Pro Thr Phe Leu Asp
515 520 525
Gly Trp Asp Lys Asn Lys Glu Glu Ala Asn Leu Gly Ile Leu Leu Ile
530 535 540
Lys Asp Asn Arg Tyr Tyr Leu Gly Ile Met Asn Thr Ser Ser Asn Lys
545 550 555 560
Ala Phe Val Asn Pro Pro Lys Ala Ile Ser Asn Asp Ile Tyr Lys Lys
565 570 575
Val Asp Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met Leu Pro Lys Val
580 585 590
Phe Phe Ala Thr Lys Asn Ile Ala Tyr Tyr Ala Pro Ser Glu Glu Leu
595 600 605
Leu Ser Lys Tyr Arg Lys Gly Thr His Lys Lys Gly Asp Ser Phe Ser
610 615 620
Ile Asp Asp Cys Arg Asn Leu Ile Asp Phe Phe Lys Ser Ser Ile Asn
625 630 635 640
Lys Asn Thr Asp Trp Ser Thr Phe Gly Phe Asn Phe Ser Asp Thr Asn
645 650 655
Ser Tyr Asn Asp Ile Ser Asp Phe Tyr Arg Glu Val Glu Lys Gln Gly
660 665 670
Tyr Lys Leu Ser Phe Thr Asp Ile Asp Ala Cys Tyr Ile Lys Asp Leu
675 680 685
Val Asp Asn Asn Glu Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe
690 695 700
Ser Pro Tyr Ser Lys Gly Lys Leu Asn Leu His Thr Leu Tyr Phe Lys
705 710 715 720
Met Leu Phe Asp Gln Arg Asn Leu Asp Asn Val Val Tyr Lys Leu Asn
725 730 735
Gly Glu Ala Glu Val Phe Tyr Arg Pro Ala Ser Ile Glu Ser Asp Glu
740 745 750
Gln Ile Ile His Lys Ser Gly Gln Asn Ile Lys Asn Lys Asn Gln Lys
755 760 765
Arg Ser Asn Cys Lys Lys Thr Ser Thr Phe Asp Tyr Asp Ile Val Lys
770 775 780
Asp Arg Arg Tyr Cys Lys Asp Lys Phe Met Leu His Leu Pro Ile Thr
785 790 795 800
Val Asn Phe Gly Thr Asn Glu Ser Gly Lys Phe Asn Glu Leu Val Asn
805 810 815
Asn Ala Ile Arg Ala Asp Lys Asp Val Asn Val Ile Gly Ile Asp Arg
820 825 830
Gly Glu Arg Asn Leu Leu Tyr Val Val Val Val Asp Pro Cys Gly Lys
835 840 845
Ile Ile Glu Gln Ile Ser Leu Asn Thr Ile Val Asp Lys Glu Tyr Asp
850 855 860
Ile Glu Thr Asp Tyr His Gln Leu Leu Asp Glu Lys Glu Gly Ser Arg
865 870 875 880
Asp Lys Ala Arg Lys Asp Trp Asn Thr Ile Glu Asn Ile Lys Glu Leu
885 890 895
Lys Glu Gly Tyr Leu Ser Gln Val Val Asn Ile Ile Ala Lys Leu Val
900 905 910
Leu Lys Tyr Asp Ala Ile Ile Cys Leu Glu Asp Leu Asn Phe Gly Phe
915 920 925
Lys Arg Gly Arg Gln Lys Val Glu Lys Gln Val Tyr Gln Lys Phe Glu
930 935 940
Lys Met Leu Ile Asp Lys Met Asn Tyr Leu Val Leu Asp Lys Ser Arg
945 950 955 960
Lys Gln Glu Ser Pro Gln Lys Pro Gly Gly Ala Leu Asn Ala Leu Gln
965 970 975
Leu Thr Ser Ala Phe Lys Ser Phe Lys Glu Leu Gly Lys Gln Thr Gly
980 985 990
Ile Ile Tyr Tyr Val Pro Ala Tyr Leu Thr Ser Lys Ile Asp Pro Thr
995 1000 1005
Thr Gly Phe Ala Asn Leu Phe Tyr Ile Lys Tyr Glu Ser Val Asp
1010 1015 1020
Lys Ala Arg Asp Phe Phe Ser Lys Phe Asp Phe Ile Arg Tyr Asn
1025 1030 1035
Gln Met Asp Asn Tyr Phe Glu Phe Gly Phe Asp Tyr Lys Ser Phe
1040 1045 1050
Thr Glu Arg Ala Ser Gly Cys Lys Ser Lys Trp Ile Ala Cys Thr
1055 1060 1065
Asn Gly Glu Arg Ile Val Lys Tyr Arg Asn Ser Asp Lys Asn Asn
1070 1075 1080
Ser Phe Asp Asp Lys Thr Val Ile Leu Thr Asp Glu Tyr Arg Ser
1085 1090 1095
Leu Phe Asp Lys Tyr Leu Gln Asn Tyr Ile Asp Glu Asp Asp Leu
1100 1105 1110
Lys Asp Gln Ile Leu Gln Ile Asp Ser Ala Asp Phe Tyr Lys Asn
1115 1120 1125
Leu Ile Lys Leu Phe Gln Leu Thr Leu Gln Met Arg Asn Ser Ser
1130 1135 1140
Ser Asp Gly Lys Arg Asp Tyr Ile Ile Ser Pro Val Lys Asn Tyr
1145 1150 1155
Arg Glu Glu Phe Phe Cys Ser Glu Phe Ser Asp Asp Thr Phe Pro
1160 1165 1170
Arg Asp Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala Arg Lys Gly
1175 1180 1185
Leu Trp Val Ile Lys Gln Ile Arg Glu Thr Lys Ser Gly Thr Lys
1190 1195 1200
Ile Asn Leu Ala Met Ser Asn Ser Glu Trp Leu Glu Tyr Ala Gln
1205 1210 1215
Cys Asn Leu Leu
1220
<210> 61
<211> 1326
<212> PRT
<213> 解糖胨普雷沃氏菌
<220>
<221> MISC_FEATURE
<222> (1)..(1326)
<223> Genbank WP_050786240 Cpf1
<400> 61
Met Lys Val Met Glu Asn Tyr Gln Glu Phe Thr Asn Leu Phe Gln Leu
1 5 10 15
Asn Lys Thr Leu Arg Phe Glu Leu Lys Pro Ile Gly Lys Thr Cys Glu
20 25 30
Leu Leu Glu Glu Gly Lys Ile Phe Ala Ser Gly Ser Phe Leu Glu Lys
35 40 45
Asp Lys Val Arg Ala Asp Asn Val Ser Tyr Val Lys Lys Glu Ile Asp
50 55 60
Lys Lys His Lys Ile Phe Ile Glu Glu Thr Leu Ser Ser Phe Ser Ile
65 70 75 80
Ser Asn Asp Leu Leu Lys Gln Tyr Phe Asp Cys Tyr Asn Glu Leu Lys
85 90 95
Ala Phe Lys Lys Asp Cys Lys Ser Asp Glu Glu Glu Val Lys Lys Thr
100 105 110
Ala Leu Arg Asn Lys Cys Thr Ser Ile Gln Arg Ala Met Arg Glu Ala
115 120 125
Ile Ser Gln Ala Phe Leu Lys Ser Pro Gln Lys Lys Leu Leu Ala Ile
130 135 140
Lys Asn Leu Ile Glu Asn Val Phe Lys Ala Asp Glu Asn Val Gln His
145 150 155 160
Phe Ser Glu Phe Thr Ser Tyr Phe Ser Gly Phe Glu Thr Asn Arg Glu
165 170 175
Asn Phe Tyr Ser Asp Glu Glu Lys Ser Thr Ser Ile Ala Tyr Arg Leu
180 185 190
Val His Asp Asn Leu Pro Ile Phe Ile Lys Asn Ile Tyr Ile Phe Glu
195 200 205
Lys Leu Lys Glu Gln Phe Asp Ala Lys Thr Leu Ser Glu Ile Phe Glu
210 215 220
Asn Tyr Lys Leu Tyr Val Ala Gly Ser Ser Leu Asp Glu Val Phe Ser
225 230 235 240
Leu Glu Tyr Phe Asn Asn Thr Leu Thr Gln Lys Gly Ile Asp Asn Tyr
245 250 255
Asn Ala Val Ile Gly Lys Ile Val Lys Glu Asp Lys Gln Glu Ile Gln
260 265 270
Gly Leu Asn Glu His Ile Asn Leu Tyr Asn Gln Lys His Lys Asp Arg
275 280 285
Arg Leu Pro Phe Phe Ile Ser Leu Lys Lys Gln Ile Leu Ser Asp Arg
290 295 300
Glu Ala Leu Ser Trp Leu Pro Asp Met Phe Lys Asn Asp Ser Glu Val
305 310 315 320
Ile Lys Ala Leu Lys Gly Phe Tyr Ile Glu Asp Gly Phe Glu Asn Asn
325 330 335
Val Leu Thr Pro Leu Ala Thr Leu Leu Ser Ser Leu Asp Lys Tyr Asn
340 345 350
Leu Asn Gly Ile Phe Ile Arg Asn Asn Glu Ala Leu Ser Ser Leu Ser
355 360 365
Gln Asn Val Tyr Arg Asn Phe Ser Ile Asp Glu Ala Ile Asp Ala Asn
370 375 380
Ala Glu Leu Gln Thr Phe Asn Asn Tyr Glu Leu Ile Ala Asn Ala Leu
385 390 395 400
Arg Ala Lys Ile Lys Lys Glu Thr Lys Gln Gly Arg Lys Ser Phe Glu
405 410 415
Lys Tyr Glu Glu Tyr Ile Asp Lys Lys Val Lys Ala Ile Asp Ser Leu
420 425 430
Ser Ile Gln Glu Ile Asn Glu Leu Val Glu Asn Tyr Val Ser Glu Phe
435 440 445
Asn Ser Asn Ser Gly Asn Met Pro Arg Lys Val Glu Asp Tyr Phe Ser
450 455 460
Leu Met Arg Lys Gly Asp Phe Gly Ser Asn Asp Leu Ile Glu Asn Ile
465 470 475 480
Lys Thr Lys Leu Ser Ala Ala Glu Lys Leu Leu Gly Thr Lys Tyr Gln
485 490 495
Glu Thr Ala Lys Asp Ile Phe Lys Lys Asp Glu Asn Ser Lys Leu Ile
500 505 510
Lys Glu Leu Leu Asp Ala Thr Lys Gln Phe Gln His Phe Ile Lys Pro
515 520 525
Leu Leu Gly Thr Gly Glu Glu Ala Asp Arg Asp Leu Val Phe Tyr Gly
530 535 540
Asp Phe Leu Pro Leu Tyr Glu Lys Phe Glu Glu Leu Thr Leu Leu Tyr
545 550 555 560
Asn Lys Val Arg Asn Arg Leu Thr Gln Lys Pro Tyr Ser Lys Asp Lys
565 570 575
Ile Arg Leu Cys Phe Asn Lys Pro Lys Leu Met Thr Gly Trp Val Asp
580 585 590
Ser Lys Thr Glu Lys Ser Asp Asn Gly Thr Gln Tyr Gly Gly Tyr Leu
595 600 605
Phe Arg Lys Lys Asn Glu Ile Gly Glu Tyr Asp Tyr Phe Leu Gly Ile
610 615 620
Ser Ser Lys Ala Gln Leu Phe Arg Lys Asn Glu Ala Val Ile Gly Asp
625 630 635 640
Tyr Glu Arg Leu Asp Tyr Tyr Gln Pro Lys Ala Asn Thr Ile Tyr Gly
645 650 655
Ser Ala Tyr Glu Gly Glu Asn Ser Tyr Lys Glu Asp Lys Lys Arg Leu
660 665 670
Asn Lys Val Ile Ile Ala Tyr Ile Glu Gln Ile Lys Gln Thr Asn Ile
675 680 685
Lys Lys Ser Ile Ile Glu Ser Ile Ser Lys Tyr Pro Asn Ile Ser Asp
690 695 700
Asp Asp Lys Val Thr Pro Ser Ser Leu Leu Glu Lys Ile Lys Lys Val
705 710 715 720
Ser Ile Asp Ser Tyr Asn Gly Ile Leu Ser Phe Lys Ser Phe Gln Ser
725 730 735
Val Asn Lys Glu Val Ile Asp Asn Leu Leu Lys Thr Ile Ser Pro Leu
740 745 750
Lys Asn Lys Ala Glu Phe Leu Asp Leu Ile Asn Lys Asp Tyr Gln Ile
755 760 765
Phe Thr Glu Val Gln Ala Val Ile Asp Glu Ile Cys Lys Gln Lys Thr
770 775 780
Phe Ile Tyr Phe Pro Ile Ser Asn Val Glu Leu Glu Lys Glu Met Gly
785 790 795 800
Asp Lys Asp Lys Pro Leu Cys Leu Phe Gln Ile Ser Asn Lys Asp Leu
805 810 815
Ser Phe Ala Lys Thr Phe Ser Ala Asn Leu Arg Lys Lys Arg Gly Ala
820 825 830
Glu Asn Leu His Thr Met Leu Phe Lys Ala Leu Met Glu Gly Asn Gln
835 840 845
Asp Asn Leu Asp Leu Gly Ser Gly Ala Ile Phe Tyr Arg Ala Lys Ser
850 855 860
Leu Asp Gly Asn Lys Pro Thr His Pro Ala Asn Glu Ala Ile Lys Cys
865 870 875 880
Arg Asn Val Ala Asn Lys Asp Lys Val Ser Leu Phe Thr Tyr Asp Ile
885 890 895
Tyr Lys Asn Arg Arg Tyr Met Glu Asn Lys Phe Leu Phe His Leu Ser
900 905 910
Ile Val Gln Asn Tyr Lys Ala Ala Asn Asp Ser Ala Gln Leu Asn Ser
915 920 925
Ser Ala Thr Glu Tyr Ile Arg Lys Ala Asp Asp Leu His Ile Ile Gly
930 935 940
Ile Asp Arg Gly Glu Arg Asn Leu Leu Tyr Tyr Ser Val Ile Asp Met
945 950 955 960
Lys Gly Asn Ile Val Glu Gln Asp Ser Leu Asn Ile Ile Arg Asn Asn
965 970 975
Asp Leu Glu Thr Asp Tyr His Asp Leu Leu Asp Lys Arg Glu Lys Glu
980 985 990
Arg Lys Ala Asn Arg Gln Asn Trp Glu Ala Val Glu Gly Ile Lys Asp
995 1000 1005
Leu Lys Lys Gly Tyr Leu Ser Gln Ala Val His Gln Ile Ala Gln
1010 1015 1020
Leu Met Leu Lys Tyr Asn Ala Ile Ile Ala Leu Glu Asp Leu Gly
1025 1030 1035
Gln Met Phe Val Thr Arg Gly Gln Lys Ile Glu Lys Ala Val Tyr
1040 1045 1050
Gln Gln Phe Glu Lys Ser Leu Val Asp Lys Leu Ser Tyr Leu Val
1055 1060 1065
Asp Lys Lys Arg Pro Tyr Asn Glu Leu Gly Gly Ile Leu Lys Ala
1070 1075 1080
Tyr Gln Leu Ala Ser Ser Ile Thr Lys Asn Asn Ser Asp Lys Gln
1085 1090 1095
Asn Gly Phe Leu Phe Tyr Val Pro Ala Trp Asn Thr Ser Lys Ile
1100 1105 1110
Asp Pro Val Thr Gly Phe Thr Asp Leu Leu Arg Pro Lys Ala Met
1115 1120 1125
Thr Ile Lys Glu Ala Gln Asp Phe Phe Gly Ala Phe Asp Asn Ile
1130 1135 1140
Ser Tyr Asn Asp Lys Gly Tyr Phe Glu Phe Glu Thr Asn Tyr Asp
1145 1150 1155
Lys Phe Lys Ile Arg Met Lys Ser Ala Gln Thr Arg Trp Thr Ile
1160 1165 1170
Cys Thr Phe Gly Asn Arg Ile Lys Arg Lys Lys Asp Lys Asn Tyr
1175 1180 1185
Trp Asn Tyr Glu Glu Val Glu Leu Thr Glu Glu Phe Lys Lys Leu
1190 1195 1200
Phe Lys Asp Ser Asn Ile Asp Tyr Glu Asn Cys Asn Leu Lys Glu
1205 1210 1215
Glu Ile Gln Asn Lys Asp Asn Arg Lys Phe Phe Asp Asp Leu Ile
1220 1225 1230
Lys Leu Leu Gln Leu Thr Leu Gln Met Arg Asn Ser Asp Asp Lys
1235 1240 1245
Gly Asn Asp Tyr Ile Ile Ser Pro Val Ala Asn Ala Glu Gly Gln
1250 1255 1260
Phe Phe Asp Ser Arg Asn Gly Asp Lys Lys Leu Pro Leu Asp Ala
1265 1270 1275
Asp Ala Asn Gly Ala Tyr Asn Ile Ala Arg Lys Gly Leu Trp Asn
1280 1285 1290
Ile Arg Gln Ile Lys Gln Thr Lys Asn Asp Lys Lys Leu Asn Leu
1295 1300 1305
Ser Ile Ser Ser Thr Glu Trp Leu Asp Phe Val Arg Glu Lys Pro
1310 1315 1320
Tyr Leu Lys
1325
<210> 62
<211> 998
<212> PRT
<213> 布氏弓形杆菌
<220>
<221> MISC_FEATURE
<222> (1)..(998)
<223> Genbank WP_052943011 Cpf1
<400> 62
Met Gly Leu Leu Glu His Leu Glu Gly Ala Ile Val Glu Asp Met Phe
1 5 10 15
Ser Leu Asp Tyr Phe Ser Leu Thr Leu Ser Gln Arg Tyr Ile Asp Ile
20 25 30
Tyr Asn Thr Met Ile Gly Gly Asn Thr Leu Ala Asp Gly Thr Lys Val
35 40 45
Gln Gly Ile Asn Glu Asn Ile Asn Ile Tyr Arg Gln Lys Asn Asn Ile
50 55 60
Asp Arg Lys Asn Leu Pro Thr Leu Lys Pro Leu His Lys Gln Leu Leu
65 70 75 80
Ser Asp Arg Glu Thr Leu Ser Trp Ile Pro Glu Ala Phe Lys Thr Lys
85 90 95
Glu Glu Val Val Gly Ala Ile Glu Asp Phe Tyr Lys Asn Asn Ile Ile
100 105 110
Ser Phe Lys Cys Cys Asp Asn Ile Val Asp Ile Thr Lys Gln Phe Ile
115 120 125
Asp Ile Phe Ser Leu Asn Glu Asp Tyr Glu Leu Asn Lys Ile Phe Ile
130 135 140
Lys Asn Asp Ile Ser Ile Thr Ser Ile Ser Gln Asp Ile Phe Lys Asp
145 150 155 160
Tyr Arg Ile Ile Lys Glu Ala Leu Trp Gln Lys His Ile Asn Glu Asn
165 170 175
Pro Lys Ala Ala Lys Ser Lys Asp Leu Thr Gly Asp Lys Glu Lys Tyr
180 185 190
Phe Ser Arg Lys Asn Ser Phe Phe Ser Phe Glu Glu Ile Ile Ser Ser
195 200 205
Leu Lys Leu Met Gly Arg Lys Ile Asp Leu Phe Ser Tyr Phe Lys Asp
210 215 220
Asn Val Glu Tyr Arg Ala His Ser Ile Glu Thr Thr Phe Ile Lys Trp
225 230 235 240
Gln Lys Asn Lys Asn Asp Lys Lys Thr Thr Lys Glu Leu Leu Asp Asn
245 250 255
Ile Leu Asn Leu Gln Arg Val Leu Lys Pro Leu Tyr Leu Lys Ala Glu
260 265 270
Val Glu Lys Asp Ile Leu Phe Tyr Ser Ile Phe Asp Ile Tyr Phe Glu
275 280 285
Ser Leu Asn Glu Ile Val Lys Leu Tyr Asn Lys Val Arg Asp Phe Glu
290 295 300
Ser Lys Lys Pro Tyr Ser Leu Glu Lys Phe Lys Leu Asn Phe Gln Asn
305 310 315 320
Ser Thr Leu Leu Ser Gly Trp Asp Val Asn Lys Glu Pro Asp Asn Thr
325 330 335
Ser Ile Leu Leu Lys Lys Asp Gly Leu Tyr Tyr Leu Gly Ile Met Asp
340 345 350
Lys Lys His Asn Arg Val Phe Lys Asn Leu Glu Ser Ser Lys Gly Gly
355 360 365
Tyr Glu Lys Ile Glu Tyr Lys Leu Leu Ser Gly Pro Asn Lys Met Leu
370 375 380
Pro Lys Val Phe Phe Ser Asn Lys Ser Ile Gly Tyr Tyr Asn Pro Ser
385 390 395 400
Pro Ala Leu Leu Glu Lys Tyr Lys Ser Gly Val His Lys Lys Gly Glu
405 410 415
Ser Phe Asp Leu Asn Phe Cys His Glu Leu Ile Asp Phe Phe Lys Ala
420 425 430
Ser Ile Asp Lys His Glu Asp Trp Lys Asn Phe Asn Phe Lys Phe Ser
435 440 445
Asp Thr Ser Glu Tyr Ala Asp Ile Ser Gly Phe Tyr Arg Glu Val Glu
450 455 460
Gln Gln Gly Tyr Lys Ile Thr Phe Lys Asn Ile Asp Glu Glu Phe Ile
465 470 475 480
Asn Thr Leu Ile Asn Glu Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn
485 490 495
Lys Asp Phe Ser Thr Phe Ser Lys Gly Thr Lys Asn Leu His Thr Leu
500 505 510
Tyr Trp Glu Met Ile Phe Asn Glu Glu Asn Leu Lys Asn Val Val Tyr
515 520 525
Lys Leu Asn Gly Glu Ala Glu Ile Phe Tyr Arg Lys Lys Ser Ile Glu
530 535 540
Tyr Ser Glu Asp Lys Met Lys Tyr Gly His His Tyr Glu Glu Leu Lys
545 550 555 560
Asp Lys Phe Asn Tyr Pro Ile Ile Lys Asp Lys Arg Phe Thr Met Asp
565 570 575
Lys Phe Gln Phe His Val Pro Ile Thr Met Asn Phe Lys Ala Thr Gly
580 585 590
Arg Ser Tyr Ile Asn Glu Glu Val Asn Asp Phe Leu Arg Gln Asn Ser
595 600 605
Lys Asp Val Lys Ile Ile Gly Ile Asn Arg Gly Glu Arg His Leu Ile
610 615 620
Tyr Leu Thr Met Ile Asn Ala Lys Gly Glu Ile Ile Gln Gln Tyr Ser
625 630 635 640
Leu Asn Glu Ile Val Asn Ser Tyr Asn Asn Lys Asn Phe Thr Val Asn
645 650 655
Tyr Asn Glu Lys Leu Ser Lys Lys Glu Gly Glu Arg Ala Ile Ala Arg
660 665 670
Glu Asn Trp Gly Val Val Glu Asn Ile Lys Glu Leu Lys Glu Gly Tyr
675 680 685
Leu Ser His Ala Ile His Thr Ile Ser Asn Leu Ile Val Glu Asn Asn
690 695 700
Ala Ile Val Val Leu Glu Asp Leu Asn Phe Glu Phe Lys Arg Glu Arg
705 710 715 720
Leu Lys Val Glu Lys Ser Ile Tyr Gln Lys Phe Glu Lys Met Leu Ile
725 730 735
Asp Lys Leu Asn Tyr Leu Val Asp Lys Lys Lys Asp Ile Asn Glu Asn
740 745 750
Gly Gly Leu Leu Lys Ala Leu Gln Leu Thr Asn Lys Phe Glu Ser Phe
755 760 765
Glu Lys Ile Gly Lys Gln Asn Gly Phe Leu Phe Phe Val Asn Ala Trp
770 775 780
Asn Ile Thr Lys Ile Cys Pro Val Thr Gly Phe Val Ser Leu Phe Asp
785 790 795 800
Thr Arg Tyr Gln Ser Val Asp Lys Ala Arg Glu Phe Phe Ser Lys Phe
805 810 815
Asp Ser Ile Lys Tyr Asn Glu Glu Lys Glu His Tyr Glu Phe Val Phe
820 825 830
Asp Tyr Ser Asn Phe Thr Asp Lys Ala Lys Asp Thr Lys Thr Lys Trp
835 840 845
Thr Val Cys Ser Tyr Gly Thr Arg Ile Lys Thr Phe Arg Asn Ser Glu
850 855 860
Lys Asn Asn Asn Trp Asp Asn Lys Thr Val Ser Pro Thr Glu Asp Leu
865 870 875 880
Ser Lys Leu Leu Lys Ser Cys Asp Arg Asp Ile Lys Glu Phe Ile Ile
885 890 895
Ser Gln Asp Lys Lys Glu Phe Phe Val Glu Leu Leu Glu Ile Phe Ser
900 905 910
Leu Ile Val Gln Met Lys Asn Ser Ile Ile Asn Ser Glu Ile Asp Tyr
915 920 925
Ile Ile Ser Pro Val Ala Asn Glu Asn Gly Glu Phe Phe Asp Ser Arg
930 935 940
Phe Ala Asn Ser Ser Leu Pro Lys Asn Ala Asp Ala Asn Ala Ala Tyr
945 950 955 960
Asn Thr Ala Arg Lys Gly Leu Met Leu Leu Glu Lys Ile Arg Asp Ser
965 970 975
Glu Ile Gly Lys Lys Ile Asp Met Lys Ile Thr Asn Thr Glu Trp Leu
980 985 990
Asn Phe Val Gln Glu Arg
995
<210> 63
<211> 1475
<212> PRT
<213> 欧氏密螺旋体内共生菌(Treponema endosymbiont of Eucomonympha sp.)
<220>
<221> MISC_FEATURE
<222> (1)..(1475)
<223> Genbank WP_062376669 Cpf1
<400> 63
Met Thr Ala Phe Glu Glu Leu Thr Asn Leu Tyr Ser Val Ser Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Lys Pro Ala Gly Lys Asp Gly Asn Thr Leu Ser
20 25 30
Ala Glu Glu Ser Ala Lys Leu Phe Lys Glu Ile Leu Asp Gln Asp Arg
35 40 45
Lys Ile Lys Asp Ala Tyr Leu Ala Leu Lys Pro Val Met Asp Thr Ile
50 55 60
His Glu Lys Ile Ile Asn Ser Ser Leu Gly Ser Asp Glu Ala Arg Gln
65 70 75 80
Ile Asp Phe Ser Ala Tyr Tyr Ile Glu Tyr Asn Lys Lys Asp Asn Glu
85 90 95
Tyr Ala Val Lys Lys Ala Glu Glu Ser Leu Arg Ala Ala Ile Arg Ser
100 105 110
Ala Phe Asp Lys Ala Ala Asn Glu Leu Ala Lys Asn Ala Gly Asn Asp
115 120 125
Glu Lys Gly Lys Pro Ile Phe Lys Lys Lys Lys Gly Lys Asp Val Gly
130 135 140
Val Glu Tyr Leu Thr Gln Ala Gly Ile Ile Lys Tyr Ile Glu Asn His
145 150 155 160
Ile Lys Thr Leu Val Pro Gln Lys Val Lys Glu Phe Ile Asp Lys Lys
165 170 175
Asn Val Ile Asn Ala Lys Gly Lys Lys Ile Thr Glu Arg Thr Gly His
180 185 190
Leu Ala Val Phe Asp Ser Phe Phe Thr Tyr Phe Gly Gly Tyr Asn Thr
195 200 205
Asn Arg Lys Asn Tyr Tyr Thr Glu Asp Lys Glu Lys Ala Thr Ala Val
210 215 220
Ala Thr Arg Ile Val His Asp Asn Leu Pro Lys Phe Cys Asp Asn Cys
225 230 235 240
Ile Gln Phe Ser Gln Asp Lys Ile Val Thr Lys Arg Lys Ser Lys Glu
245 250 255
Thr Ile Pro Ala Arg Lys Asp Glu Tyr Leu Asn Ala Tyr Gln Phe Leu
260 265 270
Lys Asp Ser Glu Lys Thr Thr Gln Ile Lys Asp Ala Ala Thr Asn Thr
275 280 285
Met Ile Glu Ala Tyr Pro Val Asp Glu Arg Thr Phe Glu Ile Ala Arg
290 295 300
Phe Ser Glu Cys Leu Thr Gln Ala Gly Ile Glu Glu Tyr Asn Arg Ile
305 310 315 320
Ile Gly His Tyr Asn Ser Leu Ile Asn Leu Tyr Asn Gln Thr Arg Lys
325 330 335
Gly Glu Ser Asp Phe Arg Lys Leu Glu Gln Phe Lys Thr Leu His Lys
340 345 350
Gln Ile Gly Cys Gly Glu Lys Lys Gly Trp Arg Asp Ala Leu Lys Asp
355 360 365
Asn Glu Asp Leu Lys Gly Lys Leu Lys Ala Ile Ser Glu Ala Gly Gln
370 375 380
Lys Tyr Phe Thr Ala Ser Leu Asn Pro Asp Asp Ile Thr Phe Phe Ser
385 390 395 400
Phe Ile Asp Trp Leu Lys Lys Asn Glu Asp Trp Asp Gly Val Tyr Trp
405 410 415
Ser Lys Ser Ala Val Asp Lys Val Ser Gly Val Tyr Phe Ala Asn Trp
420 425 430
His Asp Ile Lys Asp Arg Leu Lys Gly Asn Lys Ala Cys Val His Leu
435 440 445
Asp Lys Asn Lys Glu Leu Gln Ile Asn Asp Ala Val Glu Leu Ser Gly
450 455 460
Leu Phe Ala Val Ile Asn Ser Gly Asn Ser Glu Ser Ala Asp Ile His
465 470 475 480
Ala Gln Glu His Gly Leu Phe Asp Ala Leu Asn Gln Glu Asn Asn Glu
485 490 495
Asp Leu Ser Lys Thr Leu Phe Lys Gln Ser Val Leu Asp Glu Lys Ala
500 505 510
Ser Leu Ile Asp Glu Lys Leu Ser Ala Ser Lys Asn Leu Ile Asn Leu
515 520 525
Ile Cys Ala Asp Met Glu Asn Leu Ala Asn Ala Phe Cys Glu Thr Ser
530 535 540
Ala Glu Ile Met Lys Ile Thr Asp Phe Lys Asn Glu Ala Asn Ile Leu
545 550 555 560
Ser Ile Lys Asn Trp Leu Asp Thr Ala Lys Ser Leu Ile Trp Arg Val
565 570 575
Lys Asp Phe Asp Ile Lys Glu Ser Lys Arg Lys Gly Asn Ala Ile Asn
580 585 590
Ala Glu Leu Ser Asn Met Leu Thr Glu Leu Leu His Ala Asp Asp Ala
595 600 605
Arg Trp Phe Asp Trp Tyr Asp Leu Val Arg Asn Tyr Leu Thr Lys Lys
610 615 620
Pro Gln Asp Asp Ala Lys Ala Asn Lys Leu Lys Leu Asn Phe Gly Tyr
625 630 635 640
Gly Lys Leu Leu Asp Gly Phe Val Asp Ser His Thr Asp Glu Ser Asp
645 650 655
Ala Ser Thr Gln Tyr Gly Gly Tyr Leu Phe Arg Lys Arg Thr Asp Ala
660 665 670
Gln Glu Lys Ser Asp Phe Glu Tyr Phe Leu Gly Ile Ser Lys Asn Thr
675 680 685
Lys Leu Phe Arg Cys His Leu Gln Ser Thr Val Gln Asn His Asp Lys
690 695 700
Ser Asn Phe Glu Arg Leu Glu Tyr Tyr Gln Ala Lys Ser Thr Thr Tyr
705 710 715 720
Phe Asp Ala Lys Tyr Ser Glu Asn Lys Ala Lys Leu Val Glu Ile Ile
725 730 735
Glu Ser Leu Ile Asp Asp Arg Ala Lys Thr Asp Ser Glu Leu Thr Val
740 745 750
Ile Gly Asn Glu Ile Lys Lys Arg Asp Gly Lys Gly Glu Ile Thr Pro
755 760 765
Ser Ala Leu Phe Asp Arg Val Lys Lys Asp Lys Ala Phe Ser Arg Ile
770 775 780
Leu Asn Asp Asp Thr Leu Leu Lys Ala Val Thr Gln Ala Ile Arg Asp
785 790 795 800
Leu Gln Asn Ser Cys Asp Asn Phe Ser Glu Arg Ala Pro Arg Leu Lys
805 810 815
Glu Val Gln Lys Arg Gly Tyr Phe Gly Ile Val Gly Phe Lys Gln Ile
820 825 830
Val Glu Asp Leu Gln Thr Val Ala Lys Glu Asn Lys Val Phe Asn Phe
835 840 845
Phe Asn Val Ser Gln Pro Glu Phe Glu Asn Ala Phe Ile Ser Gly Gly
850 855 860
Lys Arg Ile Tyr Leu Phe Lys Ile Ser Asn Lys Asp Leu Ser Tyr Ser
865 870 875 880
Glu Thr Ser Gln Lys Asp Glu Asn Gly Gln Gln Lys Arg Asn Phe Lys
885 890 895
Gly Ile Glu Asn Leu His Thr His Tyr Phe Arg Ala Leu Met Arg Glu
900 905 910
Tyr Lys Asn Cys Thr Asn Val Asp Leu Gly Lys Gly Glu Ile Phe Phe
915 920 925
Arg Ala Pro Val Pro Glu Ile Lys Lys Glu Ala Thr His Lys Val Tyr
930 935 940
Asp Lys Met Val Asn Arg Arg Glu Asn Glu Thr His Ile Ala Ile Pro
945 950 955 960
Glu Lys Val His Asn Glu Leu Leu Leu Phe Ala Asn Glu Lys Ile Ala
965 970 975
Val Asn Asn Leu Ser Asn Glu Thr Lys Leu Tyr Leu Asp Gln Asn Lys
980 985 990
Lys Ile Asp Glu Ser Arg Val Lys Ile Lys Asp Val Lys His Asp Ile
995 1000 1005
Ile Lys Asp Lys Arg Phe Thr Glu Ala Lys Tyr Gln Leu His Leu
1010 1015 1020
Ser Ile Leu Leu Asn Phe Thr Pro Thr Lys Glu Glu Val Asn Ala
1025 1030 1035
Lys Ile Asn Asp Thr Phe Thr Lys Ser Asp Asp Ile Gln Phe Leu
1040 1045 1050
Gly Ile Asp Arg Gly Glu Lys His Leu Ile Tyr Tyr Ser Leu Val
1055 1060 1065
Asp Ala Asn Gly Thr Ile Arg Ala Gln Asp His Phe Asp Val Ile
1070 1075 1080
Asn Lys Thr Asp Tyr Leu Gln Lys Ile Thr Glu Ala Ala Lys Ile
1085 1090 1095
Arg Arg Glu Lys Gln Glu Asn Trp Gln Gln Lys Gly Asn Ile Ser
1100 1105 1110
Asn Leu Lys Asp Gly Tyr Ile Ser Leu Val Val His Glu Ile Ile
1115 1120 1125
Glu Lys Met Lys Asp Glu Asn Gly Ser Phe Lys Pro Met Phe Ile
1130 1135 1140
Val Leu Glu Asp Leu Asn Thr Gly Phe Lys Arg Ser Arg Gln Lys
1145 1150 1155
Phe Glu Gln Gln Val Tyr Gln Lys Phe Glu Leu Ala Leu Ala Lys
1160 1165 1170
Lys Leu Asn Tyr Leu Val Asp Lys Asn Ala Lys Asp Gly Glu Leu
1175 1180 1185
Ala Ser Val Ser Arg Ala Leu Gln Leu Thr Pro Leu Val Met Asn
1190 1195 1200
Tyr Gln Asp Ile Glu Asn Arg Lys Gln Val Gly Ile Met Leu Tyr
1205 1210 1215
Thr Arg Ala Asn Tyr Thr Ser Val Thr Asp Pro Ala Thr Gly Trp
1220 1225 1230
Arg Lys Thr Val Tyr Leu Lys Pro Gly Ser Glu Glu Ser Ile Lys
1235 1240 1245
Lys Gln Ile Leu Gly Val Phe Ser Glu Ile Gly Val Asp Glu Lys
1250 1255 1260
Gly Asp Tyr Phe Phe Gln Tyr Ser Asp Thr Asn Thr Glu Arg Thr
1265 1270 1275
Trp Arg Leu Trp Ser Ser Lys Asn Gly Lys Ser Leu Glu Arg Tyr
1280 1285 1290
Arg Gly Arg Arg Asp Lys Asn Thr Asn Ala Phe Thr Val Glu Pro
1295 1300 1305
Tyr Asp Val Lys Ala Thr Leu Asn Lys Leu Phe Val Gly Phe Asp
1310 1315 1320
Lys Asp Met Ser Leu Leu Lys Gln Leu Lys Asp Gly Lys Thr Leu
1325 1330 1335
Ser Lys Ile Asp Gly Arg Lys Glu Thr Ala Trp Glu Ser Leu Arg
1340 1345 1350
Phe Val Ile Asp Leu Ile Gln Gln Ile Arg Asn Ser Gly Asp Thr
1355 1360 1365
Ser Lys Asn Gln Asp Asp Asn Phe Leu Leu Ser Pro Val Arg Asn
1370 1375 1380
Ala Gln Gly Glu His Phe Asp Ser Arg Leu Tyr Gln Asn Gln Glu
1385 1390 1395
Thr Pro Lys Leu Pro Lys Asp Ala Asp Ala Asn Gly Ala Tyr Asn
1400 1405 1410
Ile Ala Arg Lys Gly Ile Ile Met Tyr Ala His Ile Arg Gln Trp
1415 1420 1425
Ile Asn Asp Gly Gly Gln Arg Phe Glu Lys Ser Ser Asp Leu Asp
1430 1435 1440
Leu Phe Val Ser Asp Asn Glu Trp Asp Leu Trp Leu Phe Asp Ser
1445 1450 1455
Lys Gln Trp Lys Glu Gln Leu Gln Lys Phe Ala Ser Arg Lys Gln
1460 1465 1470
Lys Lys
1475
<210> 64
<211> 1251
<212> PRT
<213> 结膜炎莫拉氏菌
<220>
<221> MISC_FEATURE
<222> (1)..(1251)
<223> Genbank WP_062499108 Cpf1
<400> 64
Met Leu Phe Gln Asp Phe Thr His Leu Tyr Pro Leu Ser Lys Thr Val
1 5 10 15
Arg Phe Glu Leu Lys Pro Ile Gly Lys Thr Leu Glu His Ile His Ala
20 25 30
Lys Asn Phe Leu Ser Gln Asp Glu Thr Met Ala Asp Met Tyr Gln Lys
35 40 45
Val Lys Ala Ile Leu Asp Asp Tyr His Arg Asp Phe Ile Thr Lys Met
50 55 60
Met Ser Glu Val Thr Leu Thr Lys Leu Pro Glu Phe Tyr Glu Val Tyr
65 70 75 80
Leu Ala Leu Arg Lys Asn Pro Lys Asp Asp Thr Leu Gln Lys Gln Leu
85 90 95
Thr Glu Ile Gln Thr Ala Leu Arg Glu Glu Val Val Lys Pro Ile Asp
100 105 110
Ser Gly Gly Lys Tyr Lys Ala Gly Tyr Glu Arg Leu Phe Gly Ala Lys
115 120 125
Leu Phe Lys Asp Gly Lys Glu Leu Gly Asp Leu Ala Lys Phe Val Ile
130 135 140
Ala Gln Glu Gly Glu Ser Ser Pro Lys Leu Pro Gln Ile Ala His Phe
145 150 155 160
Glu Lys Phe Ser Thr Tyr Phe Thr Gly Phe His Asp Asn Arg Lys Asn
165 170 175
Met Tyr Ser Ser Asp Asp Lys His Thr Ala Ile Ala Tyr Arg Leu Ile
180 185 190
His Glu Asn Leu Pro Arg Phe Ile Asp Asn Leu Gln Ile Leu Val Thr
195 200 205
Ile Lys Gln Lys His Ser Val Leu Tyr Asp Gln Ile Val Asn Glu Leu
210 215 220
Asn Ala Asn Gly Leu Asp Val Ser Leu Ala Ser His Leu Asp Gly Tyr
225 230 235 240
His Lys Leu Leu Thr Gln Glu Gly Ile Thr Ala Tyr Asn Arg Ile Ile
245 250 255
Gly Glu Val Asn Ser Tyr Thr Asn Lys His Asn Gln Ile Cys His Lys
260 265 270
Ser Glu Arg Ile Ala Lys Leu Arg Pro Leu His Lys Gln Ile Leu Ser
275 280 285
Asp Gly Met Gly Val Ser Phe Leu Pro Ser Lys Phe Ala Asp Asp Ser
290 295 300
Glu Met Cys Gln Ala Val Asn Glu Phe Tyr Arg His Tyr Ala His Val
305 310 315 320
Phe Ala Lys Val Gln Ser Leu Phe Asp Arg Phe Asp Asp Tyr Gln Lys
325 330 335
Asp Gly Ile Tyr Val Glu His Lys Asn Leu Asn Glu Leu Ser Lys Gln
340 345 350
Ala Phe Gly Asp Phe Ala Leu Leu Gly Arg Val Leu Asp Gly Tyr Tyr
355 360 365
Val Asp Val Val Asn Pro Glu Phe Asn Asp Lys Phe Ala Lys Ala Lys
370 375 380
Thr Asp Asn Ala Lys Glu Lys Leu Thr Lys Glu Lys Asp Lys Phe Ile
385 390 395 400
Lys Gly Val His Ser Leu Ala Ser Leu Glu Gln Ala Ile Glu His Tyr
405 410 415
Ile Ala Gly His Asp Asp Glu Ser Val Gln Ala Gly Lys Leu Gly Gln
420 425 430
Tyr Phe Lys His Gly Leu Ala Gly Val Asp Asn Pro Ile Gln Lys Ile
435 440 445
His Asn Ser His Ser Thr Ile Lys Gly Phe Leu Glu Arg Glu Arg Pro
450 455 460
Ala Gly Glu Arg Thr Leu Pro Lys Ile Lys Ser Asp Lys Ser Leu Glu
465 470 475 480
Met Thr Gln Leu Arg Gln Leu Lys Glu Leu Leu Asp Asn Ala Leu Asn
485 490 495
Val Val His Phe Ala Lys Leu Leu Thr Thr Lys Thr Thr Leu Asp Asn
500 505 510
Gln Asp Gly Asn Phe Tyr Gly Glu Phe Gly Ala Leu Tyr Asp Glu Leu
515 520 525
Ala Lys Ile Ala Thr Leu Tyr Asn Lys Val Arg Asp Tyr Leu Ser Gln
530 535 540
Lys Pro Phe Ser Thr Glu Lys Tyr Lys Leu Asn Phe Gly Asn Pro Thr
545 550 555 560
Leu Leu Asn Gly Trp Asp Leu Asn Lys Glu Lys Asp Asn Phe Gly Val
565 570 575
Ile Leu Gln Lys Asp Gly Cys Tyr Tyr Leu Ala Leu Leu Asp Lys Ala
580 585 590
His Lys Lys Val Phe Asp Asn Ala Pro Asn Thr Gly Lys Ser Val Tyr
595 600 605
Gln Lys Met Val Tyr Lys Leu Leu Pro Gly Ser Asn Lys Met Leu Pro
610 615 620
Lys Val Phe Phe Ala Lys Ser Asn Leu Asp Tyr Tyr Asn Pro Ser Ala
625 630 635 640
Glu Leu Leu Asp Lys Tyr Ala Gln Gly Thr His Lys Lys Gly Asp Asn
645 650 655
Phe Asn Leu Lys Asp Cys His Ala Leu Ile Asp Phe Phe Lys Ala Ser
660 665 670
Ile Asn Lys His Pro Glu Trp Gln His Phe Gly Phe Glu Phe Ser Leu
675 680 685
Thr Ser Ser Tyr Gln Asp Leu Ser Asp Phe Tyr Arg Glu Val Glu Pro
690 695 700
Gln Gly Tyr Gln Val Lys Phe Val Asp Ile Asp Ala Asp Tyr Ile Asp
705 710 715 720
Glu Leu Val Glu Gln Gly Gln Leu Tyr Leu Phe Gln Ile Tyr Asn Lys
725 730 735
Asp Phe Ser Pro Lys Ala His Gly Lys Pro Asn Leu His Thr Leu Tyr
740 745 750
Phe Lys Ala Leu Phe Ser Glu Asp Asn Leu Ala Asn Pro Ile Tyr Lys
755 760 765
Leu Asn Gly Glu Ala Glu Ile Phe Tyr Arg Lys Ala Ser Leu Asp Met
770 775 780
Asn Glu Thr Thr Ile His Arg Ala Gly Glu Val Leu Glu Asn Lys Asn
785 790 795 800
Pro Asp Asn Pro Lys Glu Arg Gln Phe Val Tyr Asp Ile Ile Lys Asp
805 810 815
Lys Arg Tyr Thr Gln Asp Lys Phe Met Leu His Val Pro Ile Thr Met
820 825 830
Asn Phe Gly Val Gln Gly Met Thr Ile Lys Glu Phe Asn Lys Lys Val
835 840 845
Asn Gln Ser Ile Gln Gln Tyr Asp Glu Val Asn Val Ile Gly Ile Asp
850 855 860
Arg Gly Glu Arg His Leu Leu Tyr Leu Thr Val Ile Asn Ser Lys Gly
865 870 875 880
Glu Ile Leu Glu Gln Arg Ser Leu Asn Asp Ile Ile Thr Thr Ser Ala
885 890 895
Asn Gly Thr Gln Met Thr Thr Pro Tyr His Lys Ile Leu Asp Lys Arg
900 905 910
Glu Ile Glu Arg Leu Asn Ala Arg Val Gly Trp Gly Glu Ile Glu Thr
915 920 925
Ile Lys Glu Leu Lys Ser Gly Tyr Leu Ser His Val Val His Gln Ile
930 935 940
Ser Gln Leu Met Leu Lys Tyr Asn Ala Ile Val Val Leu Glu Asp Leu
945 950 955 960
Asn Phe Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Ile Tyr
965 970 975
Gln Asn Phe Glu Asn Ala Leu Ile Lys Lys Leu Asn His Leu Val Leu
980 985 990
Lys Asp Lys Ala Asp Asn Glu Ile Gly Ser Tyr Lys Asn Ala Leu Gln
995 1000 1005
Leu Thr Asn Asn Phe Thr Asp Leu Lys Ser Ile Gly Lys Gln Thr
1010 1015 1020
Gly Phe Leu Phe Tyr Val Pro Ala Trp Asn Thr Ser Lys Ile Asp
1025 1030 1035
Pro Val Thr Gly Phe Val Asp Leu Leu Lys Pro Arg Tyr Glu Asn
1040 1045 1050
Ile Ala Gln Ser Gln Ala Phe Phe Asp Lys Phe Asp Lys Ile Cys
1055 1060 1065
Tyr Asn Ala Asp Lys Gly Tyr Phe Glu Phe His Ile Asp Tyr Ala
1070 1075 1080
Lys Phe Thr Asp Lys Ala Lys Asn Ser Arg Gln Ile Trp Thr Ile
1085 1090 1095
Cys Ser His Gly Asp Lys Arg Tyr Val Tyr Asp Lys Thr Ala Asn
1100 1105 1110
Gln Asn Lys Gly Ala Thr Ile Gly Ile Asn Val Asn Asp Glu Leu
1115 1120 1125
Lys Ser Leu Phe Ala Arg Tyr Arg Ile Asn Asp Lys Gln Pro Asn
1130 1135 1140
Leu Val Met Asp Ile Cys Gln Asn Asn Asp Lys Glu Phe His Lys
1145 1150 1155
Ser Leu Thr Tyr Leu Leu Lys Ala Leu Leu Ala Leu Arg Tyr Ser
1160 1165 1170
Asn Ala Ser Ser Asp Glu Asp Phe Ile Leu Ser Pro Val Ala Asn
1175 1180 1185
Asp Lys Gly Val Phe Phe Asn Ser Ala Leu Ala Asp Asp Thr Gln
1190 1195 1200
Pro Gln Asn Ala Asp Ala Asn Gly Ala Tyr His Ile Ala Leu Lys
1205 1210 1215
Gly Leu Trp Leu Leu Asn Glu Leu Lys Asn Ser Asp Asp Leu Asp
1220 1225 1230
Lys Val Lys Leu Ala Ile Asp Asn Gln Thr Trp Leu Asn Phe Ala
1235 1240 1245
Gln Asn Arg
1250
<210> 65
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的cTAG_A序列
<400> 65
actgggtgga atcccttctg cagcacctgg attaccctgt tatccctagt 50
<210> 66
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的cTAG_B序列
<400> 66
taatgagtag tcctcatctc cctcaagcag gcgccggcgg tactgccatc 50
<210> 67
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的cTAG_C序列
<400> 67
catataatct ccctcaagca ggccccgctg gcgcgcgcga atgttaggaa 50
<210> 68
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的cTAG_D序列
<400> 68
gcctataatg tgaagagctt cactgagtag ggcccgggct gtaaacggtt 50
<210> 69
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的cTAG_E序列
<400> 69
attcgctagc agatgtagtg tttccacagg ggcgatcgct gatatgggtc 50
<210> 70
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的cTAG_F序列
<400> 70
actacctagc tgcattttca ggaggaagcg atgggcggcc gcacaccttc 50
<210> 71
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的cTAG_G序列
<400> 71
tgataatggg tgagtgagtg tgtgcgtgtg gggcgcgcca gatgggaaca 50
<210> 72
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的cTAG_H序列
<400> 72
actccagtct ttctagaaga tggcaaacag ctattatggg tattatgggt 50
<210> 73
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的cTAG_I序列
<400> 73
tagtggacgg ggccactagg gacaggattg gcctgcagga ttcccgtcaa 50
<210> 74
<211> 42
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的cTAG_J序列
<400> 74
tgaactaagg cggctgcaca accagtggag gcctaaatga tc 42
<210> 75
<211> 25
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的标签D_FWD PCR引物
<400> 75
gcctataatg tgaagagctt cactg 25
<210> 76
<211> 19
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的标签E_REV PCR引物
<400> 76
gacccatatc agcgatcgc 19
<210> 77
<211> 9312
<212> DNA
<213> 人工序列
<220>
<223> 质粒13000444591
<400> 77
tccaaacgag agtctaatag aatgaggtcg aaaagtaaat cgcgcgggtt tgttactgat 60
aaagcaggca agacctaaaa tgtgtaaagg gcaaagtgta tactttggcg tcacccctta 120
catattttag gtcttttttt attgtgcgta actaacttgc catcttcaaa caggagggct 180
ggaagaagca gaccgctaac acagtacata aaaaaggaga catgaacgac tccagtcttt 240
ctagaagatg gcaaacagct attatgggta ttatgggtcc ccgaagcagg gttatgcagc 300
ggaaaagctc cccgaaaagt gccacctggg tccttttcat cacgtgctat aaaaataatt 360
ataatttaaa ttttttaata taaatatata aattaaaaat agaaagtaaa aaaagaaatt 420
aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac tctaggggga tcgccaacaa 480
atactacctt ttatcttgct cttcctgctc tcaggtatta atgccgaatt gtttcatctt 540
gtctgtgtag aagaccacac acgaaaatcc tgtgatttta cattttactt atcgttaatc 600
gaatgtatat ctatttaatc tgcttttctt gtctaataaa tatatatgta aagtacgctt 660
tttgttgaaa ttttttaaac ctttgtttat tttttttttc ttcattccgt aactcttcta 720
ccttctttat ttactttcta aaatccaaat acaaaacata aaaataaata aacacagagt 780
aaattcccaa attattccat cattaaaaga tacgaggcgc gtgtaagtta caggcaagcg 840
atccgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag 900
gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc 960
ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc 1020
gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt 1080
actgagagtg caccatacca cagccggaag aggagtaggg aatattactg gctgaaaata 1140
agtcttgaat gaacgtatac gcgtatattt ctaccaatct ctcaacactg agtaatggta 1200
gttataagaa agagaccgag ttagggacag ttagaggcgg tggagatatt ccttatggca 1260
tgtctggcga tgataaaact tttcaaacgg cagccccgat ctaaaagagc tgacagggaa 1320
atggtcagaa aaagaaacgt gcacccgccc gtctggacgc gccgctcacc cgcacggcag 1380
agaccaatca gtaaaaatca acggttaacg acattactat atatataata taggaagcat 1440
ttaatagaac agcatcgtaa tatatgtgta ctttgcagtt atgacgccag atggcagtag 1500
tggaagatat tctttattga aaaatagctt gtcaccttac gtacaatctt gatccggagc 1560
ttttcttttt ttgccgatta agaattcggt cgaaaaaaga aaaggagagg gccaagaggg 1620
agggcattgg tgactattga gcacgtgagt atacgtgatt aagcacacaa aggcagcttg 1680
gagtatgtct gttattaatt tcacaggtag ttctggtcca ttggtgaaag tttgcggctt 1740
gcagagcaca gaggccgcag aatgtgctct agattccgat gctgacttgc tgggtattat 1800
atgtgtgccc aatagaaaga gaacaattga cccggttatt gcaaggaaaa tttcaagtct 1860
tgtaaaagca tataaaaata gttcaggcac tccgaaatac ttggttggcg tgtttcgtaa 1920
tcaacctaag gaggatgttt tggctctggt caatgattac ggcattgata tcgtccaact 1980
gcatggagat gagtcgtggc aagaatacca agagttcctc ggtttgccag ttattaaaag 2040
actcgtattt ccaaaagact gcaacatact actcagtgca gcttcacaga aacctcattc 2100
gtttattccc ttgtttgatt cagaagcagg tgggacaggt gaacttttgg attggaactc 2160
gatttctgac tgggttggaa ggcaagagag ccccgaaagc ttacatttta tgttagctgg 2220
tggactgacg ccagaaaatg ttggtgatgc gcttagatta aatggcgtta ttggtgttga 2280
tgtaagcgga ggtgtggaga caaatggtgt aaaagactct aacaaaatag caaatttcgt 2340
caaaaatgct aagaaatagg ttattactga gtagtattta tttaagtatt gtttgtgcac 2400
ttgcctgcag gccttttgaa aagcaagcat aaaagatcta aacataaaat ctgtaaaata 2460
acaagatgta aagataatgc taaatcattt ggctttttga ttgattgtac aggactgggt 2520
ggaatccctt ctgcagcacc tggattaccc tgttatccct agttttgatg ggtatcggta 2580
ttgctactgg ggaaaatcgc gcggcagagg cagcaaaaaa agcaatttcc agcccgcttc 2640
ttgaagcggc cattgacggt gcgcaaggcg tcctcatgaa catcactgga ggaacaaacc 2700
tcagcctata tgaggttcag gaagcagcag acattgtcgc ttcggcgtct gatcaagacg 2760
taaacatgat tttcggttct gttattaatg aaaatctaaa agatgagatt gtggtgacag 2820
tgattgcaac cggctttatc gaacaagaga aggacgtgac gaagcctcag cgcccaagct 2880
taaatcaaag catcaaaaca cacaatcaaa gtgttccgaa gcgtgacgca aaacgtgagg 2940
aacctcagca gcagaacaca gtaagccgtc atacttcaca gccggctgat gatacgcttg 3000
acatcccgac attcttaaga aaccgtaata aacgcggcta atgtaaagga caaaatcgtt 3060
ttcgattttg tcttttttgt ttttctcttc acacttcctt cttataaagt ctttttccct 3120
attgctttcc ttcgcttagt aacaaaacag ataattagac ccatttattt ttgtgacatt 3180
tttatcattt tcatatatat ggaaattgaa tgacatgaaa cgacaatatc tgtaattcag 3240
attgtctaca gttaatatac agcgatgttc tgacaaacca ttcattatta aaaggaggga 3300
cgacactttt tttaaaaagc atgttgaaaa agggggatga aaatgaggaa aaaaacgaaa 3360
aacagactca tcagctctgt tttaagtaca gttgtcatca gttcactgct gtttccggga 3420
gcagccgggg caagcagtaa agtcacctca ccttctgtta aaaaggagct tcaatctgcg 3480
gaatccattc aaaacaagat ttcgagttca ttaaagaaaa gctttaaaaa gaaagaaaaa 3540
acgacttttc tgattaaatt taaagatctt aatgagtagt cctcatctcc ctcaagcagg 3600
cgccggcggt actgccatcc aaaaatcaga ccagacaaaa gcggcaaatg aataagcgga 3660
acggggaagg atttgcggtc aagtccttcc cttccgcacg tatcaattcg caagcttttc 3720
ctttataata gaatgaatga gaaggatgaa accgaatgaa cacttatgaa caaattaaca 3780
aagtgaaaaa gatcttacgc aaacacttga aaaataactt aattggcaca tacatgttcg 3840
gtagcggagt agaaagcggc cttaaaccga acagcgacct tgacttcctt gttgttgttt 3900
cagagccttt aacggatcaa tctaaagaaa ttttaatcca gaaaattcgc ccaatctcta 3960
aaaaaatcgg agacaaatca aatctgcgct atatcgaact tacaattatt attcaacaag 4020
agatggtccc atggaatcac cctccgaaac aagagttcat ttacggtgag tggttgcagg 4080
aactctatga acaaggatat atcccgcaga aggagcttaa ctccgatctt actatcatgc 4140
tttaccaagc gaaacgtaaa aataaacgta tctacggtaa ctacgatctc gaagaacttc 4200
ttcctgatat tccattctct gatgttcgcc gcgccatcat ggatagctct gaagaactca 4260
tcgacaacta ccaggacgat gaaacaaact ctattcttac attatgtcgc atgattctta 4320
ctatggatac gggtaaaatc atcccaaagg atattgcagg aaacgcagta gctgaatcct 4380
cccctcttga gcatcgcgaa cgcattcttc ttgctgtacg ttcttacctt ggggaaaata 4440
tcgaatggac aaacgaaaat gttaatttaa caatcaacta tcttaacaac cgccttaaaa 4500
aactgtgaaa aaaagcgcag ctgaaatagc tgcgcttttt tgtgtcataa catataatct 4560
ccctcaagca ggccccgctg gcgcgcgcga atgttaggaa acgattagtc ttttgactgt 4620
ttgacggtgg tggtactggg gcctataatg tgaagagctt cactgagtag ggcccgggct 4680
gtaaacggtt aattttgtca aaataatttt attgacaacg tcttattaac gttgatataa 4740
tttaaatttt atttgacaaa aatgggctcg tgttgtacaa taaatgttac tagagaaagg 4800
tggtgaatac tagatgacgg cattgacgga aggtgcaaaa ctgtttgaga aagagatccc 4860
gtatatcacc gaactggaag gcgacgtcga aggtatgaaa tttatcatta aaggcgaggg 4920
taccggtgac gcgaccacgg gtaccattaa agcgaaatac atctgcacta cgggcgacct 4980
gccggtcccg tgggcaaccc tggtgagcac cctgagctac ggtgttcagt gtttcgccaa 5040
gtacccgagc cacatcaagg atttctttaa gagcgccatg ccggaaggtt atacccaaga 5100
gcgtaccatc agcttcgaag gcgacggcgt gtacaagacg cgtgctatgg ttacctacga 5160
acgcggttct atctacaatc gtgtcacgct gactggtgag aactttaaga aagacggtca 5220
cattctgcgt aagaacgttg cattccaatg cccgccaagc attctgtata ttctgcctga 5280
caccgttaac aatggcatcc gcgttgagtt caaccaggcg tacgatattg aaggtgtgac 5340
cgaaaaactg gttaccaaat gcagccaaat gaatcgtccg ttggcgggct ccgcggcagt 5400
gcatatcccg cgttatcatc acattaccta ccacaccaaa ctgagcaaag accgcgacga 5460
gcgccgtgat cacatgtgtc tggtagaggt cgtgaaagcg gttgatctgg acacgtatca 5520
gtaaaaaaaa agcgcagctg aaatagctgc gcttttttgt gtcataaccc tttacagtca 5580
taaaaattat ggtataatca tttctgttgt ctttttaaag acacaagcat gaccattatg 5640
actagattcg ctagcagatg tagtgtttcc acaggggcga tcgctgatat gggtccagtt 5700
caaaagggat tatggtcgat aaagattttt atctcgtata tatccagtcg aaacctgatc 5760
cgtattcacc tggattggca atggatgaaa ccggccagaa ttccggccgc aactggcagt 5820
atatagatgg aaagtggcag ccaggtgaca aagcggatgg caactatatg attcgcgcat 5880
tagttgatta tgaagctgct gtacctgaga ttacttcacc gacagacaaa tcatacacaa 5940
ataaggatag cgtcactgta aaaggaaacg cttctcctgg cacaacggta cacatttata 6000
atggagagaa agaagcagga gaaacgaaag ctgctgcgga tggcacgttc catgcaggca 6060
tcatactcaa caagggtgaa aatgagctga cggcaactgc atcaactgac aacggaacaa 6120
cagatgcctc cagcccaatc acggtcacgc ttgatcaaga aaagcctgaa ttaacactgg 6180
acaatccaaa ggatggcggg aaaacaaata aagaaacgct gactgtcaaa ggggctgtat 6240
ccgatgacaa tctgaaagac gtcaaggtga atggcaaaaa agcaacagta gctgatggtt 6300
catactcagc ccgtattctt ttggaaaatg gaagaaatga aatcaaggta attgctacag 6360
acttggcagg caacaaaacg acgaaaaaga cagtcattga tgtgaacttt gacaagcctg 6420
tcatttccgg cttaattccg ggagaggata aaaacttaaa agccggtgaa tctgtgaaaa 6480
tcgctttctc aagcgctgag gatttagatg caacgtttac cattcgtatg ccgctgacca 6540
atgcaagagc gagtgtgcaa aatgccaccg aactcccgtt aagagaaatc tctccgggga 6600
gatatgaagg ctattggact gccacttctt ctattaaagc aaaaggagca aaagtagaag 6660
tgatcgtccg agatgattat ggaaatgaaa caagaaaaac tgcgaatgga aaacttaata 6720
tgaacacaga aaatactacc tagctgcatt ttcaggagga agcgatgggc ggccgcacac 6780
cttctatgcg gtgtgaaata ccgccatgac ccttaaatat tctgacaaat gctctttccc 6840
taaactcccc ccataaaaaa acccgccgaa gcgggttttt acgttatttg cggattaacg 6900
attactcgtt atcagaaccg cccaggatgc ctggcagttc cctactctcg ccgctgcgct 6960
cggtcgttcg gctgcgggac ctcagcgcta gcggagtgta tactggctta ctatgttggc 7020
actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc 7080
gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct 7140
cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag 7200
atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag 7260
gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc 7320
gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt 7380
tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac 7440
gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc 7500
ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa 7560
gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg 7620
aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca 7680
agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc 7740
aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa 7800
gatcatctta ttaagcctca ctcattaggc accccaggct ttacactgat aatgggtgag 7860
tgagtgtgtg cgtgtggggc gcgccagatg ggaacagcta gcttcacgct gccgcaagca 7920
ctcagggcgc aagggctgct aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg 7980
gtgctgaccc cggatgaatg tcagctactg ggctatctgg acaagggaaa acgcaagcgc 8040
aaagagaaag caggtagctt gcagtgggct tacatggcga tagctagact gggcggtttt 8100
atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc 8160
ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag 8220
atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 8280
aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 8340
cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 8400
caagaccgac ctgtccggtg ccctgaatga actccaagac gaggcagcgc ggctatcgtg 8460
gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 8520
ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 8580
tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 8640
tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 8700
agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 8760
actgttcgcc aggctcaagg cgcggatgcc cgacggcgag gatctcgtcg tgacccatgg 8820
cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 8880
tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 8940
tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 9000
cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 9060
gggttcgcta gaggatcgat cctttttaac ccatcacata tacctgccgt tcactattat 9120
ttagtgaaat gagatattat gatattttct gaattgtgat taaaaaggca actttatgcc 9180
catgcaacag aaactataaa aaatacagag aatgaaaaga aacagataga ttttttagtt 9240
ctttaggccc gtagtctgca aatcctttta tgattttcta tcaaacaaaa gaggaaaata 9300
gaccagttgc aa 9312
<210> 78
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> 从pMLN005化学合成的克隆标签cTag K
<400> 78
gcagatgtag tgtttccaca gggcccccct tattaaatga cttctcgaaa 50
<210> 79
<211> 77
<212> DNA
<213> 人工序列
<220>
<223> 从pMLN005化学合成的克隆标签cTag L
<400> 79
tttcgtctag agccttttgt ataaaaatta gggaccttgc actgactgcc ccccggtgag 60
tgagtgtgtg cgtgtgg 77
<210> 80
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> 从pMAS006化学合成的克隆标签cTag K’
<400> 80
gcagatgtag tgtttccaca gggtttcgag aagtcattta ataagccccc 50
<210> 81
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> 从pMAS006化学合成的克隆标签cTag L’
<400> 81
aaaaaataca aaaggctcta gacgaaaggt gagtgagtgt gtgcgtgtgg 50
<210> 82
<211> 2781
<212> DNA
<213> 人工序列
<220>
<223> 在体外合成地构建的质粒pMLN005 CamR
<400> 82
gcattttcag gaggaagcga tgggcggccg cacaccttct catgaccaaa atcccttaac 60
gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 120
atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 180
tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 240
gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga 300
actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 360
gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 420
agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 480
ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 540
aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 600
cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 660
gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 720
cctgataatt gtaggctgga taagatgcgt cagcatcgca tccggcaaag gcagatctcg 780
cagatgtagt gtttccacag ggcccccttg atcgggcacg taagaggttc caactttcac 840
cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct 900
aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg 960
catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc 1020
gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat 1080
ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca 1140
atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat 1200
gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt 1260
ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa 1320
gggtttattg agaatatgtt tttcgtttca gccaatccct gggtgagttt caccagtttt 1380
gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat 1440
tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt 1500
gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag 1560
ggcggggcgt aatttgatat cgagctcgct tggactcctg ttgatagatc cagtaatgac 1620
ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg 1680
taaaaattag ggaccttgca ctgactgccc cccggtgagt gagtgtgtgc gtgtggtcag 1740
aattggttaa ttggttgtaa cactgacccc tatttgttta tttttctaaa tacattcaaa 1800
tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa 1860
gaatatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct 1920
tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg 1980
tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg 2040
ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt 2100
atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga 2160
cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga 2220
attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac 2280
gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg 2340
ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac 2400
gatgcctgta gcgatggcaa caacgttgcg caaactatta actggcgaac tacttactct 2460
agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct 2520
gcgctcggcc cttccggctg gctggtttat tgctgataaa tccggagccg gtgagcgtgg 2580
ttctcgcggt atcatcgcag cgctggggcc agatggtaag ccctcccgta tcgtagttat 2640
ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg 2700
tgcctcactg attaagcatt ggtaataatg aagcgatagc gccggcttag tcagatttaa 2760
tctgcgcgcg tggtggatat t 2781
<210> 83
<211> 3128
<212> DNA
<213> 人工序列
<220>
<223> 合成地构建的质粒13000789485 pMLN 005
<400> 83
gcattttcag gaggaagcga tgggcggccg cacaccttct catgaccaaa atcccttaac 60
gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 120
atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 180
tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 240
gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga 300
actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 360
gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 420
agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 480
ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 540
aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 600
cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 660
gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 720
cctgataatt gtaggctgga taagatgcgt cagcatcgca tccggcaaag gcagatctcg 780
cagatgtagt gtttccacag ggcccccctt attaaatgac ttctcgaaac tgtggataac 840
cgtattaccg ccggcggtgt tgacataaat accactggcg gtgatactga gcacatcagc 900
aggtcacaca ggaaagtact agatgtcgca tcttgcagaa ttagtagctt cagcgaaggc 960
cgcgatttct caggcgagtg acgtcgcagc actggataat gtacgtgttg agtacctggg 1020
aaagaaggga caccttactc ttcaaatgac aaccctgcgc gaactgccgc cggaggaacg 1080
ccccgcagca ggagcggtaa tcaatgaggc aaaggagcaa gtacaacagg cactgaacgc 1140
ccgtaaggct gagttggaat ccgccgcatt aaacgcgcgc cttgctgcgg aaaccattga 1200
tgtctcgctg cccgggcgcc gcattgagaa tggaggctta cacccagtga ctcgtaccat 1260
cgaccgtatc gaatctttct ttggcgaact tggcttcact gtggcaactg gaccggagat 1320
tgaggacgac taccacaatt tcgatgcctt gaacattccc ggtcatcatc ctgcacgcgc 1380
cgatcatgat acattctggt ttgataccac ccgtttgctt cgtacccaga caagcggtgt 1440
ccaaatccgt acgatgaagg ctcagcaacc accgatccgt atcattgctc cagggcgcgt 1500
gtaccgtaac gattatgacc agacacatac accgatgttt caccaaatgg aagggttgat 1560
tgtggatacg aatatctctt tcacgaatct gaagggcacc ttacatgatt tcttacgcaa 1620
ctttttcgag gaggaccttc aaattcgctt tcgtccatcg tacttccctt ttgcagaacc 1680
ttcggctgaa gtggatgtaa tggggaaaaa cggtaagtgg ctggaggttt taggttgcgg 1740
gatggttcat ccaaatgtgc ttcgcaacgt cggcatcgac cccgaagtct acagtggatt 1800
cggattcggg atgggaatgg aacgtctgac tatgcttcgt tacggcgtaa cggatttgcg 1860
ctcctttttt gagaacgatc ttcgttttct gaagcaattc aaataagcat ttttagtacg 1920
tgcaataacc actctggttt ttccagggtg gttttttgat gccctttttg gagtcttcaa 1980
ctgctgcgtt atcccctgat tctgtgtttc gtctagagcc ttttgtataa aaattaggga 2040
ccttgcactg actgcccccc ggtgagtgag tgtgtgcgtg tggtcagaat tggttaattg 2100
gttgtaacac tgacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 2160
atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagaa tatgagtatt 2220
caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 2280
cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 2340
tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 2400
tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac 2460
gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 2520
tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 2580
gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 2640
aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 2700
gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagcg 2760
atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 2820
caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 2880
ccggctggct ggtttattgc tgataaatcc ggagccggtg agcgtggttc tcgcggtatc 2940
atcgcagcgc tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 3000
agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 3060
aagcattggt aataatgaag cgatagcgcc ggcttagtca gatttaatct gcgcgcgtgg 3120
tggatatt 3128
<210> 84
<211> 2498
<212> DNA
<213> 人工序列
<220>
<223> 合成地构建的质粒13000823784 pMAS006
<400> 84
acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg 60
cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg 120
aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag 180
gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt 240
cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca ggtcgactct 300
agaggatccc cgcagatgta gtgtttccac agggtttcga gaagtcattt aataagcccc 360
cttgatcggg cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg 420
cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat 480
cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt 540
tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt 600
aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg 660
cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg 720
ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct 780
ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc 840
gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt 900
ttcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa 960
cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat 1020
gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct 1080
taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttg atatcgagct 1140
cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca 1200
gaacgctcgg ttgccgccgg gcgtttttta ttggtaaaaa atacaaaagg ctctagacga 1260
aaggtgagtg agtgtgtgcg tgtgggtcga ctctagagga tccccgggta ccgagctcga 1320
attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta 1380
atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg 1440
atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc 1500
tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct 1560
ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac 1620
gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca 1680
tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac 1740
gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt 1800
ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt 1860
atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta 1920
tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg 1980
tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac 2040
gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg 2100
aagaacgccc tggcttgttg tccacaaccg ttaaacctta aaagctttaa aagccttata 2160
tattcttttt tttcttataa aacttaaaac cttagaggct atttaagttg ctgatttata 2220
ttaattttat tgttcaaaca tgagagctta gtacgtgaaa catgagagct tagtacgtta 2280
gccatgaggg tttagttcgt tagccatgag ggtttagttc gttaaacatg agagcttagt 2340
acgttaaaca tgagagctta gtacgtgaaa catgagagct tagtacgtac tatcaacagg 2400
ttgaactgct gatcttcttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta 2460
ccgcctttga gtgagctgat accgctcgcc gcagccga 2498
<210> 85
<211> 11567
<212> DNA
<213> 人工序列
<220>
<223> 合成地构建的质粒13000283399
<400> 85
actacctagc tgcattttca ggaggaagcg atgggcggcc gcacaccttc tatgcggtgt 60
gaaataccgc catgaccctt aaatattctg acaaatgctc tttccctaaa ctccccccat 120
aaaaaaaccc gccgaagcgg gtttttacgt tatttgcgga ttaacgatta ctcgttatca 180
gaaccgccca ggatgcctgg cagttcccta ctctcgccgc tgcgctcggt cgttcggctg 240
cgggacctca gcgctagcgg agtgtatact ggcttactat gttggcactg atgagggtgt 300
cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca gcagaatatg 360
tgatacagga tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg 420
cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata 480
cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg 540
acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa 600
gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt 660
ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt 720
tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac 780
cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca 840
ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg 900
ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg 960
gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc 1020
gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa 1080
gcctcactca ttaggcaccc caggctttac actgataatg ggtgagtgag tgtgtgcgtg 1140
tggggcgcgc cagatgggaa cagctagctt cacgctgccg caagcactca gggcgcaagg 1200
gctgctaaag gaagcggaac acgtagaaag ccagtccgca gaaacggtgc tgaccccgga 1260
tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag agaaagcagg 1320
tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg acagcaagcg 1380
aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact 1440
ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct gatcaagaga 1500
caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg 1560
cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg 1620
ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt 1680
ccggtgccct gaatgaactc caagacgagg cagcgcggct atcgtggctg gccacgacgg 1740
gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat 1800
tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat 1860
ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg 1920
accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg 1980
atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc 2040
tcaaggcgcg gatgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc 2100
cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg 2160
tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg 2220
gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca 2280
tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgctagagg 2340
atcgatcctt tttaacccat cacatatacc tgccgttcac tattatttag tgaaatgaga 2400
tattatgata ttttctgaat tgtgattaaa aaggcaactt tatgcccatg caacagaaac 2460
tataaaaaat acagagaatg aaaagaaaca gatagatttt ttagttcttt aggcccgtag 2520
tctgcaaatc cttttatgat tttctatcaa acaaaagagg aaaatagacc agttgcaatc 2580
caaacgagag tctaatagaa tgaggtcgaa aagtaaatcg cgcgggtttg ttactgataa 2640
agcaggcaag acctaaaatg tgtaaagggc aaagtgtata ctttggcgtc accccttaca 2700
tattttaggt ctttttttat tgtgcgtaac taacttgcca tcttcaaaca ggagggctgg 2760
aagaagcaga ccgctaacac agtacataaa aaaggagaca tgaacgactc cagtctttct 2820
agaagatggc aaacagctat tatgggtatt atgggtcccc gaagcagggt tatgcagcgg 2880
aaaagctccc cgaaaagtgc cacctgggtc cttttcatca cgtgctataa aaataattat 2940
aatttaaatt ttttaatata aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa 3000
agaaaaaata gtttttgttt tccgaagatg taaaagactc tagggggatc gccaacaaat 3060
actacctttt atcttgctct tcctgctctc aggtattaat gccgaattgt ttcatcttgt 3120
ctgtgtagaa gaccacacac gaaaatcctg tgattttaca ttttacttat cgttaatcga 3180
atgtatatct atttaatctg cttttcttgt ctaataaata tatatgtaaa gtacgctttt 3240
tgttgaaatt ttttaaacct ttgtttattt tttttttctt cattccgtaa ctcttctacc 3300
ttctttattt actttctaaa atccaaatac aaaacataaa aataaataaa cacagagtaa 3360
attcccaaat tattccatca ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat 3420
ccgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc 3480
cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg 3540
agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt 3600
cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac 3660
tgagagtgca ccataccaca gccggaagag gagtagggaa tattactggc tgaaaataag 3720
tcttgaatga acgtatacgc gtatatttct accaatctct caacactgag taatggtagt 3780
tataagaaag agaccgagtt agggacagtt agaggcggtg gagatattcc ttatggcatg 3840
tctggcgatg ataaaacttt tcaaacggca gccccgatct aaaagagctg acagggaaat 3900
ggtcagaaaa agaaacgtgc acccgcccgt ctggacgcgc cgctcacccg cacggcagag 3960
accaatcagt aaaaatcaac ggttaacgac attactatat atataatata ggaagcattt 4020
aatagaacag catcgtaata tatgtgtact ttgcagttat gacgccagat ggcagtagtg 4080
gaagatattc tttattgaaa aatagcttgt caccttacgt acaatcttga tccggagctt 4140
ttcttttttt gccgattaag aattcggtcg aaaaaagaaa aggagagggc caagagggag 4200
ggcattggtg actattgagc acgtgagtat acgtgattaa gcacacaaag gcagcttgga 4260
gtatgtctgt tattaatttc acaggtagtt ctggtccatt ggtgaaagtt tgcggcttgc 4320
agagcacaga ggccgcagaa tgtgctctag attccgatgc tgacttgctg ggtattatat 4380
gtgtgcccaa tagaaagaga acaattgacc cggttattgc aaggaaaatt tcaagtcttg 4440
taaaagcata taaaaatagt tcaggcactc cgaaatactt ggttggcgtg tttcgtaatc 4500
aacctaagga ggatgttttg gctctggtca atgattacgg cattgatatc gtccaactgc 4560
atggagatga gtcgtggcaa gaataccaag agttcctcgg tttgccagtt attaaaagac 4620
tcgtatttcc aaaagactgc aacatactac tcagtgcagc ttcacagaaa cctcattcgt 4680
ttattccctt gtttgattca gaagcaggtg ggacaggtga acttttggat tggaactcga 4740
tttctgactg ggttggaagg caagagagcc ccgaaagctt acattttatg ttagctggtg 4800
gactgacgcc agaaaatgtt ggtgatgcgc ttagattaaa tggcgttatt ggtgttgatg 4860
taagcggagg tgtggagaca aatggtgtaa aagactctaa caaaatagca aatttcgtca 4920
aaaatgctaa gaaataggtt attactgagt agtatttatt taagtattgt ttgtgcactt 4980
gcctgcaggc cttttgaaaa gcaagcataa aagatctaaa cataaaatct gtaaaataac 5040
aagatgtaaa gataatgcta aatcatttgg ctttttgatt gattgtacag gactgggtgg 5100
aatcccttct gcagcacctg gattaccctg ttatccctag taacaaaatt ctccagtctt 5160
cacatcggtt tgaaaggagg aagcggaaga atgaagtaag agggattttt gactccgaag 5220
taagtcttca aaaaatcaaa taaggagtgt caagaatgtt tgcaaaacga ttcaaaacct 5280
ctttactgcc gttattcgct ggatttttat tgctgtttca tttggttctg gcaggaccgg 5340
cggctgcgag tgctgaaacg gcgaacaaat cgaatgagct tacagcaccg tcgatcaaaa 5400
gcggaaccat tcttcatgca tggaattggt cgttcaatac gttaaaacac aatatgaagg 5460
atattcatga tgcaggatat acagccattc agacatctcc gattaaccaa gtaaaggaag 5520
ggaatcaagg agataaaagc atgtcgaact ggtactggct gtatcagccg acatcgtatc 5580
aaattggcaa ccgttactta ggtactgaac aagaatttaa agaaatgtgt gcagccgctg 5640
aagaatatgg cataaaggtc attgttgacg cggtcatcaa tcataccacc agtgattatg 5700
ccgcgatttc caatgaggtt aagagtattc caaactggac acatggaaac acacaaatta 5760
aaaactggtc tgatcgaaat agtacataat aatgagtagt cctcatctcc ctcaagcagg 5820
cgccggcggt actgccatct catgtttgac agcttatcat cggcaatagt tacccttatt 5880
atcaagataa gaaagaaaag gatttttcgc tacgctcaaa tcctttaaaa aaacacaaaa 5940
gaccacattt tttaatgtgg tctttattct tcaactaaag cacccattag ttcaacaaac 6000
gaaaattgga taaagtggga tatttttaaa atatatattt atgttacagt aatattgact 6060
tttaaaaaag gattgattct aatgaagaaa gcagacaagt aagcctccta aattcacttt 6120
agataaaaat ttaggaggca tatcaaatga actttaataa aattgattta gacaattgga 6180
agagaaaaga gatatttaat cattatttga accaacaaac gacttttagt ataaccacag 6240
aaattgatat tagtgtttta taccgaaaca taaaacaaga aggatataaa ttttaccctg 6300
catttatttt cttagtgaca agggtgataa actcaaatac agcttttaga actggttaca 6360
atagcgacgg agagttaggt tattgggata agttagagcc actttataca atttttgatg 6420
gtgtatctaa aacattctct ggtatttgga ctcctgtaaa gaatgacttc aaagagtttt 6480
atgatttata cctttctgat gtagagaaat ataatggttc ggggaaattg tttcccaaaa 6540
cacctatacc tgaaaatgct ttttctcttt ctattattcc atggacttca tttactgggt 6600
ttaacttaaa tatcaataat aatagtaatt accttctacc cattattaca gcaggaaaat 6660
tcattaataa aggtaattca atatatttac cgctatcttt acaggtacat cattctgttt 6720
gtgatggtta tcatgcagga ttgtttatga actctattca ggaattgtca gataggccta 6780
atgactggct tttataatat gagataatgc cgactgtact ttttacagtc ggttttctaa 6840
tgtcactaac ctgccccgtt agttgaagaa ggtttttata ttacagctcc agatcctcta 6900
cgccggacgc atcgtggcat ataatctccc tcaagcaggc cccgctggcg cgcgcgaatg 6960
ttaggaaacg attagtcttt tgactgtttg acggtggtgg tactggggcc tataatgtga 7020
agagcttcac tgagtagggc ccgggctgta aacggttgaa ttcgcggccg cttctagagg 7080
gagttctgag aattggtatg ccttataagt ccaattaaca gttgaaaacc tgcataggag 7140
agctatgcgg gttttttatt ttacataatg atacataatt taccgaaact tgcggaacat 7200
aattgaggaa tcatagaatt ttgtcaaaat aattttattg acaacgtctt attaacgttg 7260
atataattta aattttattt gacaaaaatg ggctcgtgtt gtacaataaa tgtagttact 7320
agtagcggcc gctgcaggga tccacagtac ataaaaaagg agacatgacg atggtcgttt 7380
tacaacgtga ctgggtcgac cgggaaaacc ctggcgttac ccaacttaat cgccttgcag 7440
cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc 7500
aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg 7560
tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa 7620
actggcagat gcacggttac gatgcgccca tctacaccaa cgtaacctat cccattacgg 7680
tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg 7740
ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc gttaactcgg 7800
cgtttcatct gtggtgcaac gggcgctggg tcggttacgg ccaggacagt cgtttgccgt 7860
ctgaatttga cctgagcgca tttttacgcg ccggagaaaa ccgcctcgcg gtgatggtgc 7920
tgcgttggag tgacggcagt tatctggaag atcaggatat gtggcggatg agcggcattt 7980
tccgtgacgt ctcgttgctg cataaaccga ctacacaaat cagcgatttc catgttgcca 8040
ctcgctttaa tgatgatttc agccgcgctg tactggaggc tgaagttcag atgtgcggcg 8100
agttgcgtga ctacctacgg gtaacagttt ctttatggca gggtgaaacg caggtcgcca 8160
gcggcaccgc gcctttcggc ggtgaaatta tcgatgagcg tggtggttat gccgatcgcg 8220
tcacactacg tctgaacgtc gaaaacccga aactgtggag cgccgaaatc ccgaatctct 8280
atcgtgcggt ggttgaactg cacaccgccg acggcacgct gattgaagca gaagcctgcg 8340
atgtcggttt ccgcgaggtg cggattgaaa atggtctgct gctgctgaac ggcaagccgt 8400
tgctgattcg aggcgttaac cgtcacgagc atcatcctct gcatggtcag gtcatggatg 8460
agcagacgat ggtgcaggat atcctgctga tgaagcagaa caactttaac gccgtgcgct 8520
gttcgcatta tccgaaccat ccgctgtggt acacgctgtg cgaccgctac ggcctgtatg 8580
tggtggatga agccaatatt gaaacccacg gcatggtgcc aatgaatcgt ctgaccgatg 8640
atccgcgctg gctaccggcg atgagcgaac gcgtaacgcg aatggtgcag cgcgatcgta 8700
atcacccgag tgtgatcatc tggtcgctgg ggaatgaatc aggccacggc gctaatcacg 8760
acgcgctgta tcgctggatc aaatctgtcg atccttcccg cccggtgcag tatgaaggcg 8820
gcggagccga caccacggcc accgatatta tttgcccgat gtacgcgcgc gtggatgaag 8880
accagccctt cccggctgtg ccgaaatggt ccatcaaaaa atggctttcg ctacctggag 8940
agacgcgccc gctgatcctt tgcgaatacg cccacgcgat gggtaacagt cttggcggtt 9000
tcgctaaata ctggcaggcg tttcgtcagt atccccgttt acagggcggc ttcgtctggg 9060
actgggtgga tcagtcgctg attaaatatg atgaaaacgg caacccgtgg tcggcttacg 9120
gcggtgattt tggcgatacg ccgaacgatc gccagttctg tatgaacggt ctggtctttg 9180
ccgaccgcac gccgcatcca gcgctgacgg aagcaaaaca ccagcagcag tttttccagt 9240
tccgtttatc cgggcaaacc atcgaagtga ccagcgaata cctgttccgt catagcgata 9300
acgagctcct gcactggatg gtggcgctgg atggtaagcc gctggcaagc ggtgaagtgc 9360
ctctggatgt cgctccacaa ggtaaacagt tgattgaact gcctgaacta ccgcagccgg 9420
agagcgccgg gcaactctgg ctcacagtac gcgtagtgca accgaacgcg accgcatggt 9480
cagaagccgg gcacatcagc gcctggcagc agtggcgtct ggcggaaaac ctcagtgtga 9540
cgctccccgc cgcgtcccac gccatcccgc atctgaccac cagcgaaatg gatttttgca 9600
tcgagctggg taataagcgt tggcaattta accgccagtc aggctttctt tcacagatgt 9660
ggattggcga taaaaaacaa ctgctgacgc cgctgcgcga tcagttcacc cgtgcaccgc 9720
tggataacga cattggcgta agtgaagcga cccgcattga ccctaacgcc tgggtcgaac 9780
gctggaaggc ggcgggccat taccaggccg aagcagcgtt gttgcagtgc acggcagata 9840
cacttgctga tgcggtgctg attacgaccg ctcacgcgtg gcagcatcag gggaaaacct 9900
tatttatcag ccggaaaacc taccggattg atggtagtgg tcaaatggcg attaccgttg 9960
atgttgaagt ggcgagcgat acaccgcatc cggcgcggat tggcctgaac tgccagctgg 10020
cgcaggtagc agagcgggta aactggctcg gattagggcc gcaagaaaac tatcccgacc 10080
gccttactgc cgcctgtttt gaccgctggg atctgccatt gtcagacatg tataccccgt 10140
acgtcttccc gagcgaaaac ggtctgcgct gcgggacgcg cgaattgaat tatggcccac 10200
accagtggcg cggcgacttc cagttcaaca tcagccgcta cagtcaacag caactgatgg 10260
aaaccagcca tcgccatctg ctgcacgcgg aagaaggcac atggctgaat atcgacggtt 10320
tccatatggg gattggtggc gacgactcct ggagcccgtc agtatcggcg gaattaattc 10380
cagctgagcg ccggtcgcta ccattaccag ttggtctggt gtcaaaaata ataataaccg 10440
ggcaggccat gtctgcccgt atttcgcgta aggaaatcca attcgctagc agatgtagtg 10500
tttccacagg ggcgatcgct gatatgggtc gtcgatcgac atggatgagc gatgatgata 10560
tccgtttagg ctgggcggtg atagcttctc gttcaggcag tacgcctctt ttcttttcca 10620
gacctgaggg aggcggaaat ggtgtgaggt tccccagatc tgggggaaaa gccaaatagg 10680
cgatcgcggg agtgctttat ttgaagatca ggctatcact gcggtcaata gatttcacaa 10740
tgtgatggct ggacagcctg aggaactctc gaacccgaat ggaaacaacc agatatttat 10800
gaatcagcgc ggctcacatg gcgttgtgct ggcaaatgca ggttcatcct ctgtctctat 10860
caatacggca acaaaattgc ctgatggcag gtatgacaat aaagctggag cgggttcatt 10920
tcaagtgaac gatggtaaac tgacaggcac gatcaatgcc aggtctgtag ctgtgcttta 10980
tcctgatgat attgcaaaag cgcctcatgt tttccttgag aattacaaaa caggtgtaac 11040
acattctttc aatgatcaac tgacgattac cttgcgtgca gatgcgaata caacaaaagc 11100
cgtttatcaa atcaataatg gaccagacga caggcgttta aggatggaga tcaattcaca 11160
atcggaaaag gagatccaat ttggcaaaac atacaccatc atgttaaaag gaacgaacag 11220
tgatggtgta acgaggaccg agaaatacag ttttgttaaa agagatccag cgtcggccaa 11280
aaccatcggc tatcaaaatc cgaatcattg gagccaggta aatgcttata tctataaaca 11340
tgatgggagc cgagtaattg aattgaccgg atcttggcct ggaaaaccaa tgactaaaaa 11400
tgcagacgga atttacacgc tgacgctgcc tgcggacacg gatacaacca acgcaaaagt 11460
gatttttaat aatggcagcg cccaagtgcc cggtcagaat cagcctggct ttgattacgt 11520
gctaaatggt ttatataatg actcgggctt aagcggttct cttcccc 11567
<210> 86
<211> 79
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸GFP Cpf1 cTAG M fwd
<400> 86
cagcacctgg attaccctgt tatccctagt tttgggttaa agatggttaa atgattcgaa 60
aataataaag ggaaaatca 79
<210> 87
<211> 79
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸GFP Cpf1 cTAG N fwd
<400> 87
cagcacctgg attaccctgt tatccctagt tttgggatgt taagagtccc tatcttcgaa 60
aataataaag ggaaaatca 79
<210> 88
<211> 81
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸GFP Cpf1 cTAG P fwd
<400> 88
cagcacctgg attaccctgt tatccctagt tttgaggagt gttcagtctc cgtgaactcg 60
aaaataataa agggaaaatc a 81
<210> 89
<211> 79
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸GFP Cpf1 cTAG O rvs
<400> 89
cgcttcctcc tgaaaatgca gctaggtagt tttgaccgcc ccccccatac cccaatcgac 60
atgccgaact cagaagtga 79
<210> 90
<211> 79
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸GFP Cpf1 cTAG N rvs
<400> 90
cgcttcctcc tgaaaatgca gctaggtagt tttgggatgt taagagtccc tatcttcgac 60
atgccgaact cagaagtga 79
<210> 91
<211> 51
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸CAT01 Cpf1 cTAG M fwd
<400> 91
tttgggttaa agatggttaa atgattcgac atacacataa agtagcttgc g 51
<210> 92
<211> 51
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸CAT01 Cpf1 cTAG N fwd
<400> 92
tttgggatgt taagagtccc tatcttcgac atacacataa agtagcttgc g 51
<210> 93
<211> 53
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸CAT01 Cpf1 cTAG P fwd
<400> 93
tttgaggagt gttcagtctc cgtgaactcg acatacacat aaagtagctt gcg 53
<210> 94
<211> 49
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸CAT01 Cpf1 cTAG N rvs
<400> 94
tttgggatgt taagagtccc tatcttcgac tggaaggaca agggggacc 49
<210> 95
<211> 49
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸CAT01 Cpf1 cTAG O rvs
<400> 95
tttgaccgcc ccccccatac cccaatcgac tggaaggaca agggggacc 49
<210> 96
<211> 25
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸Cpf1 cTAG M
<400> 96
tttgggttaa agatggttaa atgat 25
<210> 97
<211> 25
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸Cpf1 cTAG N
<400> 97
tttgggatgt taagagtccc tatct 25
<210> 98
<211> 25
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸Cpf1 cTAG O
<400> 98
tttgaccgcc ccccccatac cccaa 25
<210> 99
<211> 27
<212> DNA
<213> 人工序列
<220>
<223> 化学合成的寡核苷酸Cpf1 cTAG P
<400> 99
tttgaggagt gttcagtctc cgtgaac 27
<210> 100
<211> 5093
<212> DNA
<213> 人工序列
<220>
<223> 合成地构建的质粒pZHR039(卡那霉素抗性，GFP)
<400> 100
actacctagc tgcattttca ggaggaagcg atgggcggcc gcacaccttc tatgcggtgt 60
gaaataccgc catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 120
ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 180
tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 240
ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 300
tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 360
tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 420
actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 480
cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 540
gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 600
tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 660
ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 720
ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 780
cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 840
cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga 900
gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc 960
attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa 1020
ttaatgtgag ttagctcact cattaggcac cccaggcttt acactgataa tgggtgagtg 1080
agtgtgtgcg tgtggggcgc gccagatggg aacagctagc ttcacgctgc cgcaagcact 1140
cagggcgcaa gggctgctaa aggaagcgga acacgtagaa agccagtccg cagaaacggt 1200
gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa 1260
agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat 1320
ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct 1380
gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat 1440
ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 1500
gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 1560
gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 1620
agaccgacct gtccggtgcc ctgaatgaac tccaagacga ggcagcgcgg ctatcgtggc 1680
tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 1740
actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 1800
ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 1860
cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 1920
ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 1980
tgttcgccag gctcaaggcg cggatgcccg acggcgagga tctcgtcgtg acccatggcg 2040
atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 2100
gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 2160
aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 2220
attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 2280
gttcgctaga ggatcgatcc tttttaaccc atcacatata cctgccgttc actattattt 2340
agtgaaatga gatattatga tattttctga attgtgatta aaaaggcaac tttatgccca 2400
tgcaacagaa actataaaaa atacagagaa tgaaaagaaa cagatagatt ttttagttct 2460
ttaggcccgt agtctgcaaa tccttttatg attttctatc aaacaaaaga ggaaaataga 2520
ccagttgcaa tccaaacgag agtctaatag aatgaggtcg aaaagtaaat cgcgcgggtt 2580
tgttactgat aaagcaggca agacctaaaa tgtgtaaagg gcaaagtgta tactttggcg 2640
tcacccctta catattttag gtcttttttt attgtgcgta actaacttgc catcttcaaa 2700
caggagggct ggaagaagca gaccgctaac acagtacata aaaaaggaga catgaacgac 2760
tccagtcttt ctagaagatg gcaaacagct attatgggta ttatgggtcc ccgaagcagg 2820
gttatgcagc ggaaaagctc cccgaaaagt gccacctggg tccttttcat cacgtgctat 2880
aaaaataatt ataatttaaa ttttttaata taaatatata aattaaaaat agaaagtaaa 2940
aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac tctaggggga 3000
tcgccaacaa atactacctt ttatcttgct cttcctgctc tcaggtatta atgccgaatt 3060
gtttcatctt gtctgtgtag aagaccacac acgaaaatcc tgtgatttta cattttactt 3120
atcgttaatc gaatgtatat ctatttaatc tgcttttctt gtctaataaa tatatatgta 3180
aagtacgctt tttgttgaaa ttttttaaac ctttgtttat ttttttttct tcattccgta 3240
actcttctac cttctttatt tactttctaa aatccaaata caaaacataa aaataaataa 3300
acacagagta aattcccaaa ttattccatc attaaaagat acgaggcgcg tgtaagttac 3360
aggcaagcga tccgtcctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 3420
gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 3480
tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 3540
gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 3600
agcagattgt actgagagtg caccatacca cagccggaag aggagtaggg aatattactg 3660
gctgaaaata agtcttgaat gaacgtatac gcgtatattt ctaccaatct ctcaacactg 3720
agtaatggta gttataagaa agagaccgag ttagggacag ttagaggcgg tggagatatt 3780
ccttatggca tgtctggcga tgataaaact tttcaaacgg cagccccgat ctaaaagagc 3840
tgacagggaa atggtcagaa aaagaaacgt gcacccgccc gtctggacgc gccgctcacc 3900
cgcacggcag agaccaatca gtaaaaatca acggttaacg acattactat atatataata 3960
taggaagcat ttaatagaac agcatcgtaa tatatgtgta ctttgcagtt atgacgccag 4020
atggcagtag tggaagatat tctttattga aaaatagctt gtcaccttac gtacaatctt 4080
gatccggagc ttttcttttt ttgccgatta agaattcggt cgaaaaaaga aaaggagagg 4140
gccaagaggg agggcattgg tgactattga gcacgtgagt atacgtgatt aagcacacaa 4200
aggcagcttg gagtatgtct gttattaatt tcacaggtag ttctggtcca ttggtgaaag 4260
tttgcggctt gcagagcaca gaggccgcag aatgtgctct agattccgat gctgacttgc 4320
tgggtattat atgtgtgccc aatagaaaga gaacaattga cccggttatt gcaaggaaaa 4380
tttcaagtct tgtaaaagca tataaaaata gttcaggcac tccgaaatac ttggttggcg 4440
tgtttcgtaa tcaacctaag gaggatgttt tggctctggt caatgattac ggcattgata 4500
tcgtccaact gcatggagat gagtcgtggc aagaatacca agagttcctc ggtttgccag 4560
ttattaaaag actcgtattt ccaaaagact gcaacatact actcagtgca gcttcacaga 4620
aacctcattc gtttattccc ttgtttgatt cagaagcagg tgggacaggt gaacttttgg 4680
attggaactc gatttctgac tgggttggaa ggcaagagag ccccgaaagc ttacatttta 4740
tgttagctgg tggactgacg ccagaaaatg ttggtgatgc gcttagatta aatggcgtta 4800
ttggtgttga tgtaagcgga ggtgtggaga caaatggtgt aaaagactct aacaaaatag 4860
caaatttcgt caaaaatgct aagaaatagg ttattactga gtagtattta tttaagtatt 4920
gtttgtgcac ttgcctgcag gccttttgaa aagcaagcat aaaagatcta aacataaaat 4980
ctgtaaaata acaagatgta aagataatgc taaatcattt ggctttttga ttgattgtac 5040
aggactgggt ggaatccctt ctgcagcacc tggattaccc tgttatccct agt 5093
<210> 101
<211> 13737
<212> DNA
<213> 人工序列
<220>
<223> 合成地构建的质粒13000223370 Cat01(氯霉素来源)
<400> 101
gaatcttatc ccatggctaa gaaagtagac acctcgaacg ctacccccgc tctagccctt 60
cttacggaga ggcaaattcc ttttgagctg gatgttcatg atgtagatcc aaaatcatca 120
aagggcttcg cattggatgc ctctgaagta atgggtgtgg agccggaagt ggtgtttaaa 180
acgctcatgg cagatattga tggtgaacac gtggtcgcga ttgttccagc cagcagcacg 240
ttgaatctca agcagttggc taaggctgga aaaggtaagc atgcaaacat gatggatcgc 300
agccgtgcac aggtagtcac ggggtatgtc cctggtggaa tctcaccgat agggcagaag 360
aataagcacc gcgtgttttt ggatgagtct gcaattctcc aggagcgaat ctacgtcagc 420
gcaggacgcc gaggctggtc actgattatc gccccggatg atgttcttct ggctaccgat 480
ggggtttacg cggatattgc tgatcattca taaaaggtaa aacccacccc cgcaggggtg 540
cgtcgtaaag caagctcctt ttcttagtac ggcgcgatag tggactcgtg cgcgtcgatg 600
ttgatgcggc cgtgcacgga cgggaatgcg cgctgcgcgc agttctcgcg ggtgcacact 660
cggcagccgg acccgatggg ggtggcggtg gataggtcct ggaggttgaa gccgcgggag 720
tacacggtgc ggtcggcgtg gcgcgcttcg cagccgagcc cgatggcgaa cattttgtcc 780
acttcgccga accgggcttc gtggtgtcgc acggtgcgtg agatccacag gtagttgcgt 840
ccgtcgggca tttgcgcgaa ttggcggagc acttggccgg ggttggtgaa ggtttcaaac 900
acgttccaca gggggcaggt gccgccgtag tgggtgaagt ggaagccggt ggcggattgg 960
cgtttggaca tgttgccggc gcggtcgacg cgcacgaagg taaacgggat gccgcgcagg 1020
ttgggacgct gcagggtgga caggcggtgg gcggtcgtct cgtagcccac gccgaaaagc 1080
tgacctaggt attcgatgtc gtagccggat ttttcggcct cggagtggaa gattttgtag 1140
ggcagcatta cggcggcggc gaagtaggag gcgacgccgc ggatggctag ggtgcgggct 1200
tcgggggtgg accagatgcc gtcgtcgacg atgccttcga tgaggtcgtt ggcttctagg 1260
tagccgagtt cggtggccat gcggaaggcg cgttgtccgg ggttgaggcg tgcgtggatt 1320
gtcagtaggc gcgtctcggg gtcaaagtgg tgcagcgtgc cggattcctc tttggaggag 1380
gtgatggtga catcgtgatc catttgcagg cgcctggcga tcgaatcttc catggcgcgg 1440
gaatcgtacg gctgccagcc cagttgcgcg gcgatggctt cggcgcggcg gtcgagcgca 1500
tcgaagtagt tttggcgggc gtaaatgaaa tcgcgcacct cttcgtgcgg catgctcacg 1560
gcctccgcga tggggcgacg ttcctcaggc gtgttggtgc gattgtccac tgcgatggag 1620
aacttgtcgc gtacgtttcg gtagcgctgg tgcatttcca ccatggcgcg cgccagctgc 1680
gggtggttgt acaccatctc cgatagctct tggagctcca cgttcgcggg gttgatctcc 1740
cggtccaaca tgacatcctg aacttcagct aaaaggcggg aatcatcgtc gcgggagaaa 1800
aacgttgcgt ctacgccgaa cgcctcggtg atgcgcaata acaccggtac ggtgagcggg 1860
cgtacgtcgt gctcaatctg atttacataa ctggcagata agccaagggt tgctgccaac 1920
gacgcctggc tcaggtctct ttcgcggcgc agttggcgca gcctggaccc cacatatgtc 1980
tttcccatgg cgcaactata gcgtgatcac catcacctta accactcttg gcattgaggt 2040
gttgcaaacc tcagaggcta aagaacaagg gcttactgtg cgcagaggtg ccctcaccgg 2100
ggtcaaaaat ttgacatccg gtgagggtaa taccagcaca gcctagtctc gccagaatgc 2160
tggtcagcat acgaaagagc ttaaggcagg ccaattcgca ctgtcagggt cacttgggtg 2220
tttagcacta ccgacaggta cgctagtatg cgttcttcct accagaggtc tgtggccgcg 2280
tggtcaaaag tgcggctttc gtatttgctg ctcgtgttta ctctcacact tagctttgac 2340
ctgcacaaat agttgcaaat tgtcccacat acacataaag tagcttgcgt atttaaaatt 2400
atgaacctaa ggggtttagc acttcacgct gccgcaagca ctcagggcgc aagggctgct 2460
aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg 2520
tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt 2580
gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg 2640
aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg 2700
ctttcttgcc gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat 2760
gaggatcgtt tcgcatggag aaaaagatca cgggctacac taccgtggac atctcgcaat 2820
ggcatcgcaa ggaacacttc gaggcatttc aaagcgtggc acaatgtact tacaaccaga 2880
ccgtccagct ggatattacc gcgtttttga agaccgttaa gaaaaacaag cacaagtttt 2940
atccagcctt tatccatatt ctcgcccgct tgatgaatgc gcaccccgaa tttcgtatgg 3000
ccatgaaaga tggtgagctc gttatctggg actcagtcca tccatgctat accgttttcc 3060
acgaacaaac tgaaactttt tcttcgctgt ggtccgaata tcacgatgat ttccgccaat 3120
ttttgcatat ctacagccaa gatgtcgcgt gctatggtga aaacctggct tactttccca 3180
agggattcat cgagaatatg ttctttgttt cagcaaaccc ctgggtgtcc ttcacgtcgt 3240
ttgacttgaa cgtggccaat atggataatt tcttcgctcc agttttcacc atgggtaagt 3300
actataccca aggagacaag gtccttatgc cacttgcaat ccaagtacac cacgcagtct 3360
gcgatggttt ccatgtggga cgcatgctta acgaactcca acagtactgt gatgaatggc 3420
aaggcggcgc gtagcccccc aaccgaagtt gaggggattt ttgaatcctc ggtccccctt 3480
gtccttccag attgatcgac gcgttgttga ttttcgcttt tcgacgcagc ccgccgccat 3540
cgggtgcccg gcgtggtcag gccacatgcg ccccgggaac tttttgggca cctacggtgc 3600
aacagttgcg aaaattgtgt cacctgcgca aagccttgct tctattcggg aaattcgggt 3660
gtctaaactt tttggttgat accaaacggg gttagaaact gttcagatcg gtatcctgtg 3720
aggaagctca ccttggtttt agaatgttga aaaagcctca cgtttccgca ggtagagcac 3780
actcaattaa atgagcgtca aacgacaata aagtaaggct accctaataa ctggggtttt 3840
atgcctctaa acagtcagtt gggggcggta ggggagcgtc ccatgactgg ttaatgcctc 3900
gatctgggac gtacagtaac agcgacactg gaggtgccat gactgttaga aatcccgacc 3960
gtgaggcaat ccgtcacgga aaaattacga cggaggcgct gcgtgagcgt cccgcatacc 4020
cgacctgggc aatgaagctg accatggcca tcactggcct aatcttcggt ggcttcgttc 4080
ttgttcacat gatcggaaac ctgaaaatct tcatgccgga ctacgcagcc gattctgcgc 4140
atccgggtga agcacaagta gatgtctacg gcgagttcct gcgtgagatc ggatccccga 4200
tcctcccaca cggctcagtc ctctggatcc tacgtattat cctgctggtc gcattggttc 4260
tgcacatcta ctgtgcattc gcattgaccg gccgttctca ccagtcccgc ggaaagttcc 4320
gccgtaccaa cctcgttggc ggcttcaact ccttcgcgac ccgctccatg ctggtgaccg 4380
gaatcgttct ccttgcgttc attatcttcc acatcctcga cctgaccatg ggtgttgctc 4440
cagcagcccc aacttcattc gagcacggcg aagtatacgc aaatatggtg gcttccttta 4500
gccgctggcc tgtagcaatt tggtacatca ttgccaacct ggtcctgttc gtccacctgt 4560
ctcacggcat ctggcttgca gtctctgacc tgggaatcac cggacgtcgc tggagggcaa 4620
tcctcctcgc agttgcgtac atcgttcctg cactggtcct gatcggcaac atcaccattc 4680
cgttcgccat cgctgttggc tggattgcgt aaaggttagg aagaatttat gagcactcac 4740
tctgaaacca cccgcccaga gttcatccac ccagtctcag tcctcccaga ggtctcagct 4800
ggtacggtcc ttgacgctgc tgagccagca ggcgttccca ccaaagatat gtgggaatac 4860
caaaaagacc acatgaacct ggtctcccca ctgaaccgac gcaagttccg cgtcctcgtc 4920
gttggcaccg gcctgtccgg tggcgctgca gcagcagccc tcggcgaact cggatacgac 4980
gtcaaggcgt tcacctacca cgacgcacct cgccgtgcgc actccattgc tgcacagggt 5040
ggcgttaact ccgcccgcgg caagaaggta gacaacgacg gcgcataccg ccacgtcaag 5100
gacaccgtca agggcggcga ctaccgtggc cgcgagtccg actgctggcg tctcgccgtc 5160
gagtccgtcc gcgtcatcga ccacatgaac gccatcggtg caccattcgc ccgcgaatac 5220
ggtggcgcct tggcaacccg ttccttcggt ggtgtgcagg tctcccgtac ctactacacc 5280
cgtggacaaa ccggacagca gctgcagctc tccaccgcat ccgcactaca gcgccagatc 5340
cacctcggct ccgtagagat cttcacccat aacgaaatgg ttgacgtaat tgtcaccgaa 5400
cgtaatggtg aaaagcgctg cgaaggcctg atcatgcgca acctgatcac cggcgagctc 5460
accgcacaca ccggccatgc cgttatcctg gcaaccggtg gctacggcaa cgtgtaccac 5520
atgtccaccc tggcgaagaa ctccaacgcc tctagggata acagggtaat acgtcgtgac 5580
tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc 5640
tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat 5700
ggcgaatggc gataagctag cttcacgctg ccgcaagcac tcagggcgca agggctgcta 5760
aaggaagcgg aacacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt 5820
cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg 5880
cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga 5940
attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc 6000
tttcttgccg ccaaggatct gatggcgcag gggatcaaga tctgatcaag agacaggatg 6060
aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt 6120
ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt 6180
gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc 6240
cctgaatgaa ctccaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc 6300
ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga 6360
agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 6420
ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca 6480
agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga 6540
tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc 6600
gcggatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat 6660
catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 6720
ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg 6780
ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt 6840
ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgctag aggatcgatc 6900
ctttttaacc catcacatat acctgccgtt cactattatt tagtgaaatg agatattatg 6960
atattttctg aattgtgatt aaaaaggcaa ctttatgccc atgcaacaga aactataaaa 7020
aatacagaga atgaaaagaa acagatagat tttttagttc tttaggcccg tagtctgcaa 7080
atccttttat gattttctat caaacaaaag aggaaaatag accagttgca atccaaacga 7140
gagtctaata gaatgaggtc gaaaagtaaa tcgcgcgggt ttgttactga taaagcaggc 7200
aagacctaaa atgtgtaaag ggcaaagtgt atactttggc gtcacccctt acatatttta 7260
ggtctttttt tattgtgcgt aactaacttg ccatcttcaa acaggagggc tggaagaagc 7320
agaccgctaa cacagtacat aaaaaaggag acatgaacgt gagctgttta caattaatca 7380
tcgtgtggta ccatgtgtgg aattggaaag gacatgaaca tcaaaaagtt tgcaaaacaa 7440
gcaacagtat taacctttac taccgcactg ctggcaggag gcgcaactca agcgtttgcg 7500
aaagaaacga accaaaagcc atataaggaa acatacggca tttcccatat tacacgccat 7560
gatatgctgc aaatccctga acagcaaaaa aatgaaaaat atcaagtttc tgaatttgat 7620
tcgtccacaa ttaaaaatat ctcttctgca aaaggcctgg acgtttggga cagctggcca 7680
ttacaaaacg ctgacggcac tgtcgcaaac tatcacggct accacatcgt ctttgcatta 7740
gccggagatc ctaaaaatgc ggatgacaca tcgatttaca tgttctatca aaaagtcggc 7800
gaaacttcta ttgacagctg gaaaaacgct ggccgcgtct ttaaagacag cgacaaattc 7860
gatgcaaatg attctatcct aaaagaccaa acacaagaat ggtcaggttc agccacattt 7920
acatctgacg gaaaaatccg tttattctac actgatttct ccggtaaaca ttacggcaaa 7980
caaacactga caactgcaca agttaacgta tcagcatcag acagctcttt gaacatcaac 8040
ggtgtagagg attataaatc aatctttgac ggtgacggaa aaacgtatca aaatgtacag 8100
cagttcatcg atgaaggcaa ctacagctca ggcgacaacc atacgctgag agatcctcac 8160
tacgtagaag ataaaggcca caaatactta gtatttgaag caaacactgg aactgaagat 8220
ggctaccaag gcgaagaatc tttatttaac aaagcatact atggcaaaag cacatcattc 8280
ttccgtcaag aaagtcaaaa acttctgcaa agcgataaaa aacgcacggc tgagttagca 8340
aacggcgctc tcggtatgat tgagctaaac gatgattaca cactgaaaaa agtgatgaaa 8400
ccgctgattg catctaacac agtaacagat gaaattgaac gcgcgaacgt ctttaaaatg 8460
aacggcaaat ggtacctgtt cactgactcc cgcggatcaa aaatgacgat tgacggcatt 8520
acgtctaacg atatttacat gcttggttat gtttctaatt ctttaactgg cccatacaag 8580
ccgctgaaca aaactggcct tgtgttaaaa atggatcttg atcctaacga tgtaaccttt 8640
acttactcac acttcgctgt acctcaagcg aaaggaaaca atgtcgtgat tacaagctat 8700
atgacaaaca gaggattcta cgcagacaaa caatcaacgt ttgcgccgag cttcctgctg 8760
aacatcaaag gcaagaaaac atctgttgtc aaagacagca tccttgaaca aggacaatta 8820
acagttaaca aataaaaacg caaaagaaaa tgccgatggg taccgagcga aatgaccgac 8880
caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg 8940
ttgggcttcg gaatcgtttt ccgggacgcc ctcgcggacg tgctcatagt ccaataacta 9000
cattgagcga aatgccaacc acatgtccca tgcttttact aatgtggggt cttagaagaa 9060
agcgaccaat ttaaggagag ttgaatatgt ctgaaatcca gctgacggag gcatcattga 9120
acgaagcagc cgatgctgca attaaagcgt tcgatggagc acaaaacctc gatgaattgg 9180
ctgctctgcg acgagatcac ctgggtgatg cggcaccaat ccctcaggca cgccgctcgc 9240
ttggaaccat tccaaaagat cagcgtaagg atgccggacg attcgtaaac atggcgctgg 9300
gccgcgcgga aaagcacttc gcccaggtta aggtggttct tgaagaaaag cgaaacgcag 9360
aagtcctgga gctggagcgc gtggatgtta ccgtccctac cacacgtgaa caagtcggcg 9420
cactgcaccc aattacgatt ctcaacgaac agatcgcgga catctttgtt ggcatgggct 9480
gggagatcgc agagggcccg gaagttgaag ccgaatactt caatttcgat gcacttaact 9540
ttctcccaga ccacccagcc cgcaccctgc aggatacctt ccacatcgca cctgaaggat 9600
cgcgccaagt gttgcgcacc catacctctc ctgtccaggt tcgcacgatg ctgaatcgag 9660
aggtacctat ctatatcgcc tgtcctggtc gcgtcttccg cactgacgaa ttggatgcta 9720
cccacacccc tgtctttcac cagatcgagg gcctggctgt cgacaaaggc ctgacaatgg 9780
ctcaccttcg cggaactctg gatcacttgg ctaaagaact gttcggacct gagactaaaa 9840
cccgcatgcg ttcaaactac ttcccatttt ctgagcccag cgcggaagtt gatgtctggt 9900
tcccaaataa gaagggcggt gccggctgga tcgaatgggg cggctgcggc atggtcaacc 9960
caaacgtgct ccgcgctgta ggcgtcgacc cggaagaata cactggattc ggcttcggta 10020
tgggtattga acgcaccttg caattccgaa atggactctc agatatgcgc gatatggtag 10080
aaggcgacat tcgctttacc ctccctttcg gcattcaggc ttaggcattt ttagtacgtg 10140
caataaccac tctggttttt ccagggtggt tttttgatgc cctttttgga gtcttcaact 10200
gagcctcgca gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag 10260
aaggaacacc cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg 10320
atacaccaag gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag 10380
gatggatata ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaag 10440
ctccccgaaa agtgccacct gggtcctttt catcacgtgc tataaaaata attataattt 10500
aaatttttta atataaatat ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa 10560
aaatagtttt tgttttccga agatgtaaaa gactctaggg ggatcgccaa caaatactac 10620
cttttatctt gctcttcctg ctctcaggta ttaatgccga attgtttcat cttgtctgtg 10680
tagaagacca cacacgaaaa tcctgtgatt ttacatttta cttatcgtta atcgaatgta 10740
tatctattta atctgctttt cttgtctaat aaatatatat gtaaagtacg ctttttgttg 10800
aaatttttta aacctttgtt tatttttttt tcttcattcc gtaactcttc taccttcttt 10860
atttactttc taaaatccaa atacaaaaca taaaaataaa taaacacaga gtaaattccc 10920
aaattattcc atcattaaaa gatacgaggc gcgtgtaagt tacaggcaag cgatccgtcc 10980
taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt 11040
cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg 11100
gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg 11160
ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga 11220
gtgcaccata ccacagccgg aagaggagta gggaatatta ctggctgaaa ataagtcttg 11280
aatgaacgta tacgcgtata tttctaccaa tctctcaaca ctgagtaatg gtagttataa 11340
gaaagagacc gagttaggga cagttagagg cggtggagat attccttatg gcatgtctgg 11400
cgatgataaa acttttcaaa cggcagcccc gatctaaaag agctgacagg gaaatggtca 11460
gaaaaagaaa cgtgcacccg cccgtctgga cgcgccgctc acccgcacgg cagagaccaa 11520
tcagtaaaaa tcaacggtta acgacattac tatatatata atataggaag catttaatag 11580
aacagcatcg taatatatgt gtactttgca gttatgacgc cagatggcag tagtggaaga 11640
tattctttat tgaaaaatag cttgtcacct tacgtacaat cttgatccgg agcttttctt 11700
tttttgccga ttaagaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat 11760
tggtgactat tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg 11820
tctgttatta atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc 11880
acagaggccg cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg 11940
cccaatagaa agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa 12000
gcatataaaa atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct 12060
aaggaggatg ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga 12120
gatgagtcgt ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta 12180
tttccaaaag actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt 12240
cccttgtttg attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct 12300
gactgggttg gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg 12360
acgccagaaa atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc 12420
ggaggtgtgg agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat 12480
gctaagaaat aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgcctg 12540
caggcctttt gaaaagcaag cataaaagat ctaaacataa aatctgtaaa ataacaagat 12600
gtaaagataa tgctaaatca tttggctttt tgattgattg tacaggtatg cggtgtgaaa 12660
taccgccatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 12720
agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 12780
aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 12840
ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta 12900
gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 12960
aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 13020
aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 13080
gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 13140
aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 13200
aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 13260
cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 13320
cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 13380
tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 13440
tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 13500
ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta 13560
atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa 13620
tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat 13680
gttgtgtgga attgtgagcg gataacaatt taactataac ggtcctaagg tagcgaa 13737
<210> 102
<211> 6351
<212> DNA
<213> 人工序列
<220>
<223> 合成地构建的质粒pJDI427
<400> 102
ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 60
agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 120
cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 180
caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 240
tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 300
ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 360
ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 420
gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 480
gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 540
tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 600
cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 660
gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 720
ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat 780
acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 840
tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta 900
ggcaccccag gctttacact gataatgggt gagtgagtgt gtgcgtgtgg ggcgcgccag 960
atgggaacag ctagcttcac gctgccgcaa gcactcaggg cgcaagggct gctaaaggaa 1020
gcggaacacg tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta 1080
ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg 1140
gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc 1200
agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt 1260
gccgccaagg atctgatggc gcaggggatc aagatctgat caagagacag gatgaggatc 1320
gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 1380
gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 1440
gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 1500
tgaactccaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 1560
agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 1620
ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 1680
tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 1740
acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 1800
ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcggat 1860
gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 1920
ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 1980
tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 2040
ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 2100
ccttcttgac gagttcttct gagcgggact ctggggttcg ctagaggatc gatccttttt 2160
aacccatcac atatacctgc cgttcactat tatttagtga aatgagatat tatgatattt 2220
tctgaattgt gattaaaaag gcaactttat gcccatgcaa cagaaactat aaaaaataca 2280
gagaatgaaa agaaacagat agatttttta gttctttagg cccgtagtct gcaaatcctt 2340
ttatgatttt ctatcaaaca aaagaggaaa atagaccagt tgcaatccaa acgagagtct 2400
aatagaatga ggtcgaaaag taaatcgcgc gggtttgtta ctgataaagc aggcaagacc 2460
taaaatgtgt aaagggcaaa gtgtatactt tggcgtcacc ccttacatat tttaggtctt 2520
tttttattgt gcgtaactaa cttgccatct tcaaacagga gggctggaag aagcagaccg 2580
ctaacacagt acataaaaaa ggagacatga acgactccag tctttctaga agatggcaaa 2640
cagctattat gggtattatg ggtccccgaa gcagggttat gcagcggaaa agctccccga 2700
aaagtgccac ctgggtcctt ttcatcacgt gctataaaaa taattataat ttaaattttt 2760
taatataaat atataaatta aaaatagaaa gtaaaaaaag aaattaaaga aaaaatagtt 2820
tttgttttcc gaagatgtaa aagactctag ggggatcgcc aacaaatact accttttatc 2880
ttgctcttcc tgctctcagg tattaatgcc gaattgtttc atcttgtctg tgtagaagac 2940
cacacacgaa aatcctgtga ttttacattt tacttatcgt taatcgaatg tatatctatt 3000
taatctgctt ttcttgtcta ataaatatat atgtaaagta cgctttttgt tgaaattttt 3060
taaacctttg tttatttttt tttcttcatt ccgtaactct tctaccttct ttatttactt 3120
tctaaaatcc aaatacaaaa cataaaaata aataaacaca gagtaaattc ccaaattatt 3180
ccatcattaa aagatacgag gcgcgtgtaa gttacaggca agcgatccgt cctaagaaac 3240
cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc 3300
gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc 3360
ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg 3420
cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca 3480
taccacagcc ggaagaggag tagggaatat tactggctga aaataagtct tgaatgaacg 3540
tatacgcgta tatttctacc aatctctcaa cactgagtaa tggtagttat aagaaagaga 3600
ccgagttagg gacagttaga ggcggtggag atattcctta tggcatgtct ggcgatgata 3660
aaacttttca aacggcagcc ccgatctaaa agagctgaca gggaaatggt cagaaaaaga 3720
aacgtgcacc cgcccgtctg gacgcgccgc tcacccgcac ggcagagacc aatcagtaaa 3780
aatcaacggt taacgacatt actatatata taatatagga agcatttaat agaacagcat 3840
cgtaatatat gtgtactttg cagttatgac gccagatggc agtagtggaa gatattcttt 3900
attgaaaaat agcttgtcac cttacgtaca atcttgatcc ggagcttttc tttttttgcc 3960
gattaagaat tcggtcgaaa aaagaaaagg agagggccaa gagggagggc attggtgact 4020
attgagcacg tgagtatacg tgattaagca cacaaaggca gcttggagta tgtctgttat 4080
taatttcaca ggtagttctg gtccattggt gaaagtttgc ggcttgcaga gcacagaggc 4140
cgcagaatgt gctctagatt ccgatgctga cttgctgggt attatatgtg tgcccaatag 4200
aaagagaaca attgacccgg ttattgcaag gaaaatttca agtcttgtaa aagcatataa 4260
aaatagttca ggcactccga aatacttggt tggcgtgttt cgtaatcaac ctaaggagga 4320
tgttttggct ctggtcaatg attacggcat tgatatcgtc caactgcatg gagatgagtc 4380
gtggcaagaa taccaagagt tcctcggttt gccagttatt aaaagactcg tatttccaaa 4440
agactgcaac atactactca gtgcagcttc acagaaacct cattcgttta ttcccttgtt 4500
tgattcagaa gcaggtggga caggtgaact tttggattgg aactcgattt ctgactgggt 4560
tggaaggcaa gagagccccg aaagcttaca ttttatgtta gctggtggac tgacgccaga 4620
aaatgttggt gatgcgctta gattaaatgg cgttattggt gttgatgtaa gcggaggtgt 4680
ggagacaaat ggtgtaaaag actctaacaa aatagcaaat ttcgtcaaaa atgctaagaa 4740
ataggttatt actgagtagt atttatttaa gtattgtttg tgcacttgcc tgcaggcctt 4800
ttgaaaagca agcataaaag atctaaacat aaaatctgta aaataacaag atgtaaagat 4860
aatgctaaat catttggctt tttgattgat tgtacaggac tgggtggaat cccttctgca 4920
gcacctggat taccctgtta tccctagttt tgggttaaag atggttaaat gattcgaaaa 4980
taataaaggg aaaatcagtt tttgatatca aaattataca tgtcaacgat aatacaaaat 5040
ataatacaaa ctataagatg ttatcagtat ttattatgca tttagaataa attttgtgtc 5100
gcccttaatt gtgagcggat aacaattacg agcttcatgc acagtgaaat catgaaaaat 5160
ttatttgctt tgtgagcgga taacaattat aatatgtgga attgtgagcg ctcacaattc 5220
cacaacggtt tccctctaga aataattttg tttaactttt cgagacctta ggaggtaaac 5280
atatgacggc attgacggaa ggtgcaaaac tgtttgagaa agagatcccg tatatcaccg 5340
aactggaagg cgacgtcgaa ggtatgaaat ttatcattaa aggcgagggt accggtgacg 5400
cgaccacggg taccattaaa gcgaaataca tctgcactac gggcgacctg ccggtcccgt 5460
gggcaaccct ggtgagcacc ctgagctacg gtgttcagtg tttcgccaag tacccgagcc 5520
acatcaagga tttctttaag agcgccatgc cggaaggtta tacccaagag cgtaccatca 5580
gcttcgaagg cgacggcgtg tacaagacgc gtgctatggt tacctacgaa cgcggttcta 5640
tctacaatcg tgtcacgctg actggtgaga actttaagaa agacggtcac attctgcgta 5700
agaacgttgc attccaatgc ccgccaagca ttctgtatat tctgcctgac accgttaaca 5760
atggcatccg cgttgagttc aaccaggcgt acgatattga aggtgtgacc gaaaaactgg 5820
ttaccaaatg cagccaaatg aatcgtccgt tggcgggctc cgcggcagtg catatcccgc 5880
gttatcatca cattacctac cacaccaaac tgagcaaaga ccgcgacgag cgccgtgatc 5940
acatgtgtct ggtagaggtc gtgaaagcgg ttgatctgga cacgtatcag taatgagaat 6000
tctgtacact cgagggtctc accccaaggg cgacaccccc taattagccc gggcgaaagg 6060
cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgca 6120
tggggagtcc ccacactacc atcggcgcta cggcgtttca cttctgagtt cggcatgtcg 6180
aagataggga ctcttaacat cccaaaacta cctagctgca ttttcaggag gaagcgatgg 6240
gcggccgcac accttctatg cggtgtgaaa taccgccatg accaaaatcc cttaacgtga 6300
gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt c 6351
<210> 103
<211> 6351
<212> DNA
<213> 人工序列
<220>
<223> 合成地构建的质粒pJDI429
<400> 103
ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 60
agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 120
cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 180
caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 240
tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 300
ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 360
ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 420
gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 480
gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 540
tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 600
cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 660
gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 720
ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat 780
acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 840
tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta 900
ggcaccccag gctttacact gataatgggt gagtgagtgt gtgcgtgtgg ggcgcgccag 960
atgggaacag ctagcttcac gctgccgcaa gcactcaggg cgcaagggct gctaaaggaa 1020
gcggaacacg tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta 1080
ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg 1140
gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc 1200
agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt 1260
gccgccaagg atctgatggc gcaggggatc aagatctgat caagagacag gatgaggatc 1320
gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 1380
gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 1440
gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 1500
tgaactccaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 1560
agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 1620
ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 1680
tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 1740
acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 1800
ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcggat 1860
gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 1920
ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 1980
tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 2040
ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 2100
ccttcttgac gagttcttct gagcgggact ctggggttcg ctagaggatc gatccttttt 2160
aacccatcac atatacctgc cgttcactat tatttagtga aatgagatat tatgatattt 2220
tctgaattgt gattaaaaag gcaactttat gcccatgcaa cagaaactat aaaaaataca 2280
gagaatgaaa agaaacagat agatttttta gttctttagg cccgtagtct gcaaatcctt 2340
ttatgatttt ctatcaaaca aaagaggaaa atagaccagt tgcaatccaa acgagagtct 2400
aatagaatga ggtcgaaaag taaatcgcgc gggtttgtta ctgataaagc aggcaagacc 2460
taaaatgtgt aaagggcaaa gtgtatactt tggcgtcacc ccttacatat tttaggtctt 2520
tttttattgt gcgtaactaa cttgccatct tcaaacagga gggctggaag aagcagaccg 2580
ctaacacagt acataaaaaa ggagacatga acgactccag tctttctaga agatggcaaa 2640
cagctattat gggtattatg ggtccccgaa gcagggttat gcagcggaaa agctccccga 2700
aaagtgccac ctgggtcctt ttcatcacgt gctataaaaa taattataat ttaaattttt 2760
taatataaat atataaatta aaaatagaaa gtaaaaaaag aaattaaaga aaaaatagtt 2820
tttgttttcc gaagatgtaa aagactctag ggggatcgcc aacaaatact accttttatc 2880
ttgctcttcc tgctctcagg tattaatgcc gaattgtttc atcttgtctg tgtagaagac 2940
cacacacgaa aatcctgtga ttttacattt tacttatcgt taatcgaatg tatatctatt 3000
taatctgctt ttcttgtcta ataaatatat atgtaaagta cgctttttgt tgaaattttt 3060
taaacctttg tttatttttt tttcttcatt ccgtaactct tctaccttct ttatttactt 3120
tctaaaatcc aaatacaaaa cataaaaata aataaacaca gagtaaattc ccaaattatt 3180
ccatcattaa aagatacgag gcgcgtgtaa gttacaggca agcgatccgt cctaagaaac 3240
cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc 3300
gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc 3360
ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg 3420
cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca 3480
taccacagcc ggaagaggag tagggaatat tactggctga aaataagtct tgaatgaacg 3540
tatacgcgta tatttctacc aatctctcaa cactgagtaa tggtagttat aagaaagaga 3600
ccgagttagg gacagttaga ggcggtggag atattcctta tggcatgtct ggcgatgata 3660
aaacttttca aacggcagcc ccgatctaaa agagctgaca gggaaatggt cagaaaaaga 3720
aacgtgcacc cgcccgtctg gacgcgccgc tcacccgcac ggcagagacc aatcagtaaa 3780
aatcaacggt taacgacatt actatatata taatatagga agcatttaat agaacagcat 3840
cgtaatatat gtgtactttg cagttatgac gccagatggc agtagtggaa gatattcttt 3900
attgaaaaat agcttgtcac cttacgtaca atcttgatcc ggagcttttc tttttttgcc 3960
gattaagaat tcggtcgaaa aaagaaaagg agagggccaa gagggagggc attggtgact 4020
attgagcacg tgagtatacg tgattaagca cacaaaggca gcttggagta tgtctgttat 4080
taatttcaca ggtagttctg gtccattggt gaaagtttgc ggcttgcaga gcacagaggc 4140
cgcagaatgt gctctagatt ccgatgctga cttgctgggt attatatgtg tgcccaatag 4200
aaagagaaca attgacccgg ttattgcaag gaaaatttca agtcttgtaa aagcatataa 4260
aaatagttca ggcactccga aatacttggt tggcgtgttt cgtaatcaac ctaaggagga 4320
tgttttggct ctggtcaatg attacggcat tgatatcgtc caactgcatg gagatgagtc 4380
gtggcaagaa taccaagagt tcctcggttt gccagttatt aaaagactcg tatttccaaa 4440
agactgcaac atactactca gtgcagcttc acagaaacct cattcgttta ttcccttgtt 4500
tgattcagaa gcaggtggga caggtgaact tttggattgg aactcgattt ctgactgggt 4560
tggaaggcaa gagagccccg aaagcttaca ttttatgtta gctggtggac tgacgccaga 4620
aaatgttggt gatgcgctta gattaaatgg cgttattggt gttgatgtaa gcggaggtgt 4680
ggagacaaat ggtgtaaaag actctaacaa aatagcaaat ttcgtcaaaa atgctaagaa 4740
ataggttatt actgagtagt atttatttaa gtattgtttg tgcacttgcc tgcaggcctt 4800
ttgaaaagca agcataaaag atctaaacat aaaatctgta aaataacaag atgtaaagat 4860
aatgctaaat catttggctt tttgattgat tgtacaggac tgggtggaat cccttctgca 4920
gcacctggat taccctgtta tccctagttt tgggatgtta agagtcccta tcttcgaaaa 4980
taataaaggg aaaatcagtt tttgatatca aaattataca tgtcaacgat aatacaaaat 5040
ataatacaaa ctataagatg ttatcagtat ttattatgca tttagaataa attttgtgtc 5100
gcccttaatt gtgagcggat aacaattacg agcttcatgc acagtgaaat catgaaaaat 5160
ttatttgctt tgtgagcgga taacaattat aatatgtgga attgtgagcg ctcacaattc 5220
cacaacggtt tccctctaga aataattttg tttaactttt cgagacctta ggaggtaaac 5280
atatgacggc attgacggaa ggtgcaaaac tgtttgagaa agagatcccg tatatcaccg 5340
aactggaagg cgacgtcgaa ggtatgaaat ttatcattaa aggcgagggt accggtgacg 5400
cgaccacggg taccattaaa gcgaaataca tctgcactac gggcgacctg ccggtcccgt 5460
gggcaaccct ggtgagcacc ctgagctacg gtgttcagtg tttcgccaag tacccgagcc 5520
acatcaagga tttctttaag agcgccatgc cggaaggtta tacccaagag cgtaccatca 5580
gcttcgaagg cgacggcgtg tacaagacgc gtgctatggt tacctacgaa cgcggttcta 5640
tctacaatcg tgtcacgctg actggtgaga actttaagaa agacggtcac attctgcgta 5700
agaacgttgc attccaatgc ccgccaagca ttctgtatat tctgcctgac accgttaaca 5760
atggcatccg cgttgagttc aaccaggcgt acgatattga aggtgtgacc gaaaaactgg 5820
ttaccaaatg cagccaaatg aatcgtccgt tggcgggctc cgcggcagtg catatcccgc 5880
gttatcatca cattacctac cacaccaaac tgagcaaaga ccgcgacgag cgccgtgatc 5940
acatgtgtct ggtagaggtc gtgaaagcgg ttgatctgga cacgtatcag taatgagaat 6000
tctgtacact cgagggtctc accccaaggg cgacaccccc taattagccc gggcgaaagg 6060
cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgca 6120
tggggagtcc ccacactacc atcggcgcta cggcgtttca cttctgagtt cggcatgtcg 6180
attggggtat ggggggggcg gtcaaaacta cctagctgca ttttcaggag gaagcgatgg 6240
gcggccgcac accttctatg cggtgtgaaa taccgccatg accaaaatcc cttaacgtga 6300
gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt c 6351
<210> 104
<211> 6353
<212> DNA
<213> 人工序列
<220>
<223> 合成地构建的质粒pJDI430
<400> 104
ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 60
agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 120
cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 180
caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 240
tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 300
ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 360
ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 420
gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 480
gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 540
tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 600
cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 660
gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 720
ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat 780
acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 840
tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta 900
ggcaccccag gctttacact gataatgggt gagtgagtgt gtgcgtgtgg ggcgcgccag 960
atgggaacag ctagcttcac gctgccgcaa gcactcaggg cgcaagggct gctaaaggaa 1020
gcggaacacg tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta 1080
ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg 1140
gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc 1200
agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt 1260
gccgccaagg atctgatggc gcaggggatc aagatctgat caagagacag gatgaggatc 1320
gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 1380
gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 1440
gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 1500
tgaactccaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 1560
agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 1620
ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 1680
tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 1740
acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 1800
ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcggat 1860
gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 1920
ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 1980
tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 2040
ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 2100
ccttcttgac gagttcttct gagcgggact ctggggttcg ctagaggatc gatccttttt 2160
aacccatcac atatacctgc cgttcactat tatttagtga aatgagatat tatgatattt 2220
tctgaattgt gattaaaaag gcaactttat gcccatgcaa cagaaactat aaaaaataca 2280
gagaatgaaa agaaacagat agatttttta gttctttagg cccgtagtct gcaaatcctt 2340
ttatgatttt ctatcaaaca aaagaggaaa atagaccagt tgcaatccaa acgagagtct 2400
aatagaatga ggtcgaaaag taaatcgcgc gggtttgtta ctgataaagc aggcaagacc 2460
taaaatgtgt aaagggcaaa gtgtatactt tggcgtcacc ccttacatat tttaggtctt 2520
tttttattgt gcgtaactaa cttgccatct tcaaacagga gggctggaag aagcagaccg 2580
ctaacacagt acataaaaaa ggagacatga acgactccag tctttctaga agatggcaaa 2640
cagctattat gggtattatg ggtccccgaa gcagggttat gcagcggaaa agctccccga 2700
aaagtgccac ctgggtcctt ttcatcacgt gctataaaaa taattataat ttaaattttt 2760
taatataaat atataaatta aaaatagaaa gtaaaaaaag aaattaaaga aaaaatagtt 2820
tttgttttcc gaagatgtaa aagactctag ggggatcgcc aacaaatact accttttatc 2880
ttgctcttcc tgctctcagg tattaatgcc gaattgtttc atcttgtctg tgtagaagac 2940
cacacacgaa aatcctgtga ttttacattt tacttatcgt taatcgaatg tatatctatt 3000
taatctgctt ttcttgtcta ataaatatat atgtaaagta cgctttttgt tgaaattttt 3060
taaacctttg tttatttttt tttcttcatt ccgtaactct tctaccttct ttatttactt 3120
tctaaaatcc aaatacaaaa cataaaaata aataaacaca gagtaaattc ccaaattatt 3180
ccatcattaa aagatacgag gcgcgtgtaa gttacaggca agcgatccgt cctaagaaac 3240
cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc 3300
gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc 3360
ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg 3420
cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca 3480
taccacagcc ggaagaggag tagggaatat tactggctga aaataagtct tgaatgaacg 3540
tatacgcgta tatttctacc aatctctcaa cactgagtaa tggtagttat aagaaagaga 3600
ccgagttagg gacagttaga ggcggtggag atattcctta tggcatgtct ggcgatgata 3660
aaacttttca aacggcagcc ccgatctaaa agagctgaca gggaaatggt cagaaaaaga 3720
aacgtgcacc cgcccgtctg gacgcgccgc tcacccgcac ggcagagacc aatcagtaaa 3780
aatcaacggt taacgacatt actatatata taatatagga agcatttaat agaacagcat 3840
cgtaatatat gtgtactttg cagttatgac gccagatggc agtagtggaa gatattcttt 3900
attgaaaaat agcttgtcac cttacgtaca atcttgatcc ggagcttttc tttttttgcc 3960
gattaagaat tcggtcgaaa aaagaaaagg agagggccaa gagggagggc attggtgact 4020
attgagcacg tgagtatacg tgattaagca cacaaaggca gcttggagta tgtctgttat 4080
taatttcaca ggtagttctg gtccattggt gaaagtttgc ggcttgcaga gcacagaggc 4140
cgcagaatgt gctctagatt ccgatgctga cttgctgggt attatatgtg tgcccaatag 4200
aaagagaaca attgacccgg ttattgcaag gaaaatttca agtcttgtaa aagcatataa 4260
aaatagttca ggcactccga aatacttggt tggcgtgttt cgtaatcaac ctaaggagga 4320
tgttttggct ctggtcaatg attacggcat tgatatcgtc caactgcatg gagatgagtc 4380
gtggcaagaa taccaagagt tcctcggttt gccagttatt aaaagactcg tatttccaaa 4440
agactgcaac atactactca gtgcagcttc acagaaacct cattcgttta ttcccttgtt 4500
tgattcagaa gcaggtggga caggtgaact tttggattgg aactcgattt ctgactgggt 4560
tggaaggcaa gagagccccg aaagcttaca ttttatgtta gctggtggac tgacgccaga 4620
aaatgttggt gatgcgctta gattaaatgg cgttattggt gttgatgtaa gcggaggtgt 4680
ggagacaaat ggtgtaaaag actctaacaa aatagcaaat ttcgtcaaaa atgctaagaa 4740
ataggttatt actgagtagt atttatttaa gtattgtttg tgcacttgcc tgcaggcctt 4800
ttgaaaagca agcataaaag atctaaacat aaaatctgta aaataacaag atgtaaagat 4860
aatgctaaat catttggctt tttgattgat tgtacaggac tgggtggaat cccttctgca 4920
gcacctggat taccctgtta tccctagttt tgaggagtgt tcagtctccg tgaactcgaa 4980
aataataaag ggaaaatcag tttttgatat caaaattata catgtcaacg ataatacaaa 5040
atataataca aactataaga tgttatcagt atttattatg catttagaat aaattttgtg 5100
tcgcccttaa ttgtgagcgg ataacaatta cgagcttcat gcacagtgaa atcatgaaaa 5160
atttatttgc tttgtgagcg gataacaatt ataatatgtg gaattgtgag cgctcacaat 5220
tccacaacgg tttccctcta gaaataattt tgtttaactt ttcgagacct taggaggtaa 5280
acatatgacg gcattgacgg aaggtgcaaa actgtttgag aaagagatcc cgtatatcac 5340
cgaactggaa ggcgacgtcg aaggtatgaa atttatcatt aaaggcgagg gtaccggtga 5400
cgcgaccacg ggtaccatta aagcgaaata catctgcact acgggcgacc tgccggtccc 5460
gtgggcaacc ctggtgagca ccctgagcta cggtgttcag tgtttcgcca agtacccgag 5520
ccacatcaag gatttcttta agagcgccat gccggaaggt tatacccaag agcgtaccat 5580
cagcttcgaa ggcgacggcg tgtacaagac gcgtgctatg gttacctacg aacgcggttc 5640
tatctacaat cgtgtcacgc tgactggtga gaactttaag aaagacggtc acattctgcg 5700
taagaacgtt gcattccaat gcccgccaag cattctgtat attctgcctg acaccgttaa 5760
caatggcatc cgcgttgagt tcaaccaggc gtacgatatt gaaggtgtga ccgaaaaact 5820
ggttaccaaa tgcagccaaa tgaatcgtcc gttggcgggc tccgcggcag tgcatatccc 5880
gcgttatcat cacattacct accacaccaa actgagcaaa gaccgcgacg agcgccgtga 5940
tcacatgtgt ctggtagagg tcgtgaaagc ggttgatctg gacacgtatc agtaatgaga 6000
attctgtaca ctcgagggtc tcaccccaag ggcgacaccc cctaattagc ccgggcgaaa 6060
ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg 6120
catggggagt ccccacacta ccatcggcgc tacggcgttt cacttctgag ttcggcatgt 6180
cgaagatagg gactcttaac atcccaaaac tacctagctg cattttcagg aggaagcgat 6240
gggcggccgc acaccttcta tgcggtgtga aataccgcca tgaccaaaat cccttaacgt 6300
gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttc 6353
<210> 105
<211> 6353
<212> DNA
<213> 人工序列
<220>
<223> 合成地构建的质粒pJDI431
<400> 105
ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 60
agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 120
cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 180
caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 240
tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 300
ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 360
ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 420
gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 480
gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 540
tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 600
cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 660
gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 720
ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat 780
acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 840
tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta 900
ggcaccccag gctttacact gataatgggt gagtgagtgt gtgcgtgtgg ggcgcgccag 960
atgggaacag ctagcttcac gctgccgcaa gcactcaggg cgcaagggct gctaaaggaa 1020
gcggaacacg tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta 1080
ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg 1140
gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc 1200
agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt 1260
gccgccaagg atctgatggc gcaggggatc aagatctgat caagagacag gatgaggatc 1320
gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 1380
gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 1440
gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 1500
tgaactccaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 1560
agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 1620
ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 1680
tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 1740
acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 1800
ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcggat 1860
gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 1920
ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 1980
tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 2040
ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 2100
ccttcttgac gagttcttct gagcgggact ctggggttcg ctagaggatc gatccttttt 2160
aacccatcac atatacctgc cgttcactat tatttagtga aatgagatat tatgatattt 2220
tctgaattgt gattaaaaag gcaactttat gcccatgcaa cagaaactat aaaaaataca 2280
gagaatgaaa agaaacagat agatttttta gttctttagg cccgtagtct gcaaatcctt 2340
ttatgatttt ctatcaaaca aaagaggaaa atagaccagt tgcaatccaa acgagagtct 2400
aatagaatga ggtcgaaaag taaatcgcgc gggtttgtta ctgataaagc aggcaagacc 2460
taaaatgtgt aaagggcaaa gtgtatactt tggcgtcacc ccttacatat tttaggtctt 2520
tttttattgt gcgtaactaa cttgccatct tcaaacagga gggctggaag aagcagaccg 2580
ctaacacagt acataaaaaa ggagacatga acgactccag tctttctaga agatggcaaa 2640
cagctattat gggtattatg ggtccccgaa gcagggttat gcagcggaaa agctccccga 2700
aaagtgccac ctgggtcctt ttcatcacgt gctataaaaa taattataat ttaaattttt 2760
taatataaat atataaatta aaaatagaaa gtaaaaaaag aaattaaaga aaaaatagtt 2820
tttgttttcc gaagatgtaa aagactctag ggggatcgcc aacaaatact accttttatc 2880
ttgctcttcc tgctctcagg tattaatgcc gaattgtttc atcttgtctg tgtagaagac 2940
cacacacgaa aatcctgtga ttttacattt tacttatcgt taatcgaatg tatatctatt 3000
taatctgctt ttcttgtcta ataaatatat atgtaaagta cgctttttgt tgaaattttt 3060
taaacctttg tttatttttt tttcttcatt ccgtaactct tctaccttct ttatttactt 3120
tctaaaatcc aaatacaaaa cataaaaata aataaacaca gagtaaattc ccaaattatt 3180
ccatcattaa aagatacgag gcgcgtgtaa gttacaggca agcgatccgt cctaagaaac 3240
cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc 3300
gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc 3360
ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg 3420
cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca 3480
taccacagcc ggaagaggag tagggaatat tactggctga aaataagtct tgaatgaacg 3540
tatacgcgta tatttctacc aatctctcaa cactgagtaa tggtagttat aagaaagaga 3600
ccgagttagg gacagttaga ggcggtggag atattcctta tggcatgtct ggcgatgata 3660
aaacttttca aacggcagcc ccgatctaaa agagctgaca gggaaatggt cagaaaaaga 3720
aacgtgcacc cgcccgtctg gacgcgccgc tcacccgcac ggcagagacc aatcagtaaa 3780
aatcaacggt taacgacatt actatatata taatatagga agcatttaat agaacagcat 3840
cgtaatatat gtgtactttg cagttatgac gccagatggc agtagtggaa gatattcttt 3900
attgaaaaat agcttgtcac cttacgtaca atcttgatcc ggagcttttc tttttttgcc 3960
gattaagaat tcggtcgaaa aaagaaaagg agagggccaa gagggagggc attggtgact 4020
attgagcacg tgagtatacg tgattaagca cacaaaggca gcttggagta tgtctgttat 4080
taatttcaca ggtagttctg gtccattggt gaaagtttgc ggcttgcaga gcacagaggc 4140
cgcagaatgt gctctagatt ccgatgctga cttgctgggt attatatgtg tgcccaatag 4200
aaagagaaca attgacccgg ttattgcaag gaaaatttca agtcttgtaa aagcatataa 4260
aaatagttca ggcactccga aatacttggt tggcgtgttt cgtaatcaac ctaaggagga 4320
tgttttggct ctggtcaatg attacggcat tgatatcgtc caactgcatg gagatgagtc 4380
gtggcaagaa taccaagagt tcctcggttt gccagttatt aaaagactcg tatttccaaa 4440
agactgcaac atactactca gtgcagcttc acagaaacct cattcgttta ttcccttgtt 4500
tgattcagaa gcaggtggga caggtgaact tttggattgg aactcgattt ctgactgggt 4560
tggaaggcaa gagagccccg aaagcttaca ttttatgtta gctggtggac tgacgccaga 4620
aaatgttggt gatgcgctta gattaaatgg cgttattggt gttgatgtaa gcggaggtgt 4680
ggagacaaat ggtgtaaaag actctaacaa aatagcaaat ttcgtcaaaa atgctaagaa 4740
ataggttatt actgagtagt atttatttaa gtattgtttg tgcacttgcc tgcaggcctt 4800
ttgaaaagca agcataaaag atctaaacat aaaatctgta aaataacaag atgtaaagat 4860
aatgctaaat catttggctt tttgattgat tgtacaggac tgggtggaat cccttctgca 4920
gcacctggat taccctgtta tccctagttt tgaggagtgt tcagtctccg tgaactcgaa 4980
aataataaag ggaaaatcag tttttgatat caaaattata catgtcaacg ataatacaaa 5040
atataataca aactataaga tgttatcagt atttattatg catttagaat aaattttgtg 5100
tcgcccttaa ttgtgagcgg ataacaatta cgagcttcat gcacagtgaa atcatgaaaa 5160
atttatttgc tttgtgagcg gataacaatt ataatatgtg gaattgtgag cgctcacaat 5220
tccacaacgg tttccctcta gaaataattt tgtttaactt ttcgagacct taggaggtaa 5280
acatatgacg gcattgacgg aaggtgcaaa actgtttgag aaagagatcc cgtatatcac 5340
cgaactggaa ggcgacgtcg aaggtatgaa atttatcatt aaaggcgagg gtaccggtga 5400
cgcgaccacg ggtaccatta aagcgaaata catctgcact acgggcgacc tgccggtccc 5460
gtgggcaacc ctggtgagca ccctgagcta cggtgttcag tgtttcgcca agtacccgag 5520
ccacatcaag gatttcttta agagcgccat gccggaaggt tatacccaag agcgtaccat 5580
cagcttcgaa ggcgacggcg tgtacaagac gcgtgctatg gttacctacg aacgcggttc 5640
tatctacaat cgtgtcacgc tgactggtga gaactttaag aaagacggtc acattctgcg 5700
taagaacgtt gcattccaat gcccgccaag cattctgtat attctgcctg acaccgttaa 5760
caatggcatc cgcgttgagt tcaaccaggc gtacgatatt gaaggtgtga ccgaaaaact 5820
ggttaccaaa tgcagccaaa tgaatcgtcc gttggcgggc tccgcggcag tgcatatccc 5880
gcgttatcat cacattacct accacaccaa actgagcaaa gaccgcgacg agcgccgtga 5940
tcacatgtgt ctggtagagg tcgtgaaagc ggttgatctg gacacgtatc agtaatgaga 6000
attctgtaca ctcgagggtc tcaccccaag ggcgacaccc cctaattagc ccgggcgaaa 6060
ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg 6120
catggggagt ccccacacta ccatcggcgc tacggcgttt cacttctgag ttcggcatgt 6180
cgattggggt atgggggggg cggtcaaaac tacctagctg cattttcagg aggaagcgat 6240
gggcggccgc acaccttcta tgcggtgtga aataccgcca tgaccaaaat cccttaacgt 6300
gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttc 6353
<210> 106
<211> 4155
<212> DNA
<213> 人工序列
<220>
<223> 合成地构建的质粒pJDI432
<400> 106
gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc 60
ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc 120
acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact 180
cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt 240
ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc 300
ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt 360
ttcagcaaga ttttgggtta aagatggtta aatgattcga catacacata aagtagcttg 420
cgtatttaaa attatgaacc taaggggttt agcacttcac gctgccgcaa gcactcaggg 480
cgcaagggct gctaaaggaa gcggaacacg tagaaagcca gtccgcagaa acggtgctga 540
ccccggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga 600
aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt tttatggaca 660
gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa gccctgcaaa 720
gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc aagatctgat 780
caagagacag gatgaggatc gtttcgcatg gagaaaaaga tcacgggcta cactaccgtg 840
gacatctcgc aatggcatcg caaggaacac ttcgaggcat ttcaaagcgt ggcacaatgt 900
acttacaacc agaccgtcca gctggatatt accgcgtttt tgaagaccgt taagaaaaac 960
aagcacaagt tttatccagc ctttatccat attctcgccc gcttgatgaa tgcgcacccc 1020
gaatttcgta tggccatgaa agatggtgag ctcgttatct gggactcagt ccatccatgc 1080
tataccgttt tccacgaaca aactgaaact ttttcttcgc tgtggtccga atatcacgat 1140
gatttccgcc aatttttgca tatctacagc caagatgtcg cgtgctatgg tgaaaacctg 1200
gcttactttc ccaagggatt catcgagaat atgttctttg tttcagcaaa cccctgggtg 1260
tccttcacgt cgtttgactt gaacgtggcc aatatggata atttcttcgc tccagttttc 1320
accatgggta agtactatac ccaaggagac aaggtcctta tgccacttgc aatccaagta 1380
caccacgcag tctgcgatgg tttccatgtg ggacgcatgc ttaacgaact ccaacagtac 1440
tgtgatgaat ggcaaggcgg cgcgtagccc cccaaccgaa gttgagggga tttttgaatc 1500
ctcggtcccc cttgtccttc cagtcgaaga tagggactct taacatccca aaatctttct 1560
agaagatctc ctacaatatt ctcagctgcc atggaaaatc gatgttcttc ttttattctc 1620
tcaagatttt caggctgtat attaaaactt atattaagaa ctatgctaac cacctcatca 1680
ggaaccgttg taggtggcgt gggttttctt ggcaatcgac tctcatgaaa actacgagct 1740
aaatattcaa tatgttcctc ttgaccaact ttattctgca ttttttttga acgaggttta 1800
gagcaagctt caggaaactg agacaggaat tttattaaaa atttaaattt tgaagaaagt 1860
tcagggttaa tagcatccat tttttgcttt gcaagttcct cagcattctt aacaaaagac 1920
gtctcttttg acatgtttaa agtttaaacc tcctgtgtga aattattatc cgctcataat 1980
tccacacatt atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 2040
ctaactcaca ttaattgcgt tgcgctcact gccaattgct ttccagtcgg gaaacctgtc 2100
gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 2160
ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 2220
atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 2280
gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 2340
gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 2400
gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 2460
gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 2520
aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 2580
ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 2640
taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 2700
tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 2760
gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 2820
taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 2880
tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 2940
tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 3000
ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 3060
taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 3120
tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 3180
cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 3240
gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 3300
cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 3360
ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 3420
aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 3480
atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 3540
tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 3600
gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 3660
aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 3720
acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 3780
ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 3840
tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 3900
aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 3960
catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 4020
atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 4080
aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 4140
gcgtatcacg aggcc 4155
<210> 107
<211> 4155
<212> DNA
<213> 人工序列
<220>
<223> 合成地构建的质粒pJDI434
<400> 107
gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc 60
ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc 120
acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact 180
cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt 240
ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc 300
ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt 360
ttcagcaaga ttttgggatg ttaagagtcc ctatcttcga catacacata aagtagcttg 420
cgtatttaaa attatgaacc taaggggttt agcacttcac gctgccgcaa gcactcaggg 480
cgcaagggct gctaaaggaa gcggaacacg tagaaagcca gtccgcagaa acggtgctga 540
ccccggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga 600
aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt tttatggaca 660
gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa gccctgcaaa 720
gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc aagatctgat 780
caagagacag gatgaggatc gtttcgcatg gagaaaaaga tcacgggcta cactaccgtg 840
gacatctcgc aatggcatcg caaggaacac ttcgaggcat ttcaaagcgt ggcacaatgt 900
acttacaacc agaccgtcca gctggatatt accgcgtttt tgaagaccgt taagaaaaac 960
aagcacaagt tttatccagc ctttatccat attctcgccc gcttgatgaa tgcgcacccc 1020
gaatttcgta tggccatgaa agatggtgag ctcgttatct gggactcagt ccatccatgc 1080
tataccgttt tccacgaaca aactgaaact ttttcttcgc tgtggtccga atatcacgat 1140
gatttccgcc aatttttgca tatctacagc caagatgtcg cgtgctatgg tgaaaacctg 1200
gcttactttc ccaagggatt catcgagaat atgttctttg tttcagcaaa cccctgggtg 1260
tccttcacgt cgtttgactt gaacgtggcc aatatggata atttcttcgc tccagttttc 1320
accatgggta agtactatac ccaaggagac aaggtcctta tgccacttgc aatccaagta 1380
caccacgcag tctgcgatgg tttccatgtg ggacgcatgc ttaacgaact ccaacagtac 1440
tgtgatgaat ggcaaggcgg cgcgtagccc cccaaccgaa gttgagggga tttttgaatc 1500
ctcggtcccc cttgtccttc cagtcgattg gggtatgggg ggggcggtca aaatctttct 1560
agaagatctc ctacaatatt ctcagctgcc atggaaaatc gatgttcttc ttttattctc 1620
tcaagatttt caggctgtat attaaaactt atattaagaa ctatgctaac cacctcatca 1680
ggaaccgttg taggtggcgt gggttttctt ggcaatcgac tctcatgaaa actacgagct 1740
aaatattcaa tatgttcctc ttgaccaact ttattctgca ttttttttga acgaggttta 1800
gagcaagctt caggaaactg agacaggaat tttattaaaa atttaaattt tgaagaaagt 1860
tcagggttaa tagcatccat tttttgcttt gcaagttcct cagcattctt aacaaaagac 1920
gtctcttttg acatgtttaa agtttaaacc tcctgtgtga aattattatc cgctcataat 1980
tccacacatt atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 2040
ctaactcaca ttaattgcgt tgcgctcact gccaattgct ttccagtcgg gaaacctgtc 2100
gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 2160
ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 2220
atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 2280
gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 2340
gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 2400
gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 2460
gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 2520
aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 2580
ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 2640
taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 2700
tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 2760
gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 2820
taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 2880
tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 2940
tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 3000
ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 3060
taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 3120
tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 3180
cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 3240
gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 3300
cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 3360
ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 3420
aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 3480
atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 3540
tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 3600
gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 3660
aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 3720
acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 3780
ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 3840
tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 3900
aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 3960
catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 4020
atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 4080
aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 4140
gcgtatcacg aggcc 4155
<210> 108
<211> 4157
<212> DNA
<213> 人工序列
<220>
<223> 合成地构建的质粒pJDI435
<400> 108
gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc 60
ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc 120
acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact 180
cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt 240
ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc 300
ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt 360
ttcagcaaga ttttgaggag tgttcagtct ccgtgaactc gacatacaca taaagtagct 420
tgcgtattta aaattatgaa cctaaggggt ttagcacttc acgctgccgc aagcactcag 480
ggcgcaaggg ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag aaacggtgct 540
gaccccggat gaatgtcagc tactgggcta tctggacaag ggaaaacgca agcgcaaaga 600
gaaagcaggt agcttgcagt gggcttacat ggcgatagct agactgggcg gttttatgga 660
cagcaagcga accggaattg ccagctgggg cgccctctgg taaggttggg aagccctgca 720
aagtaaactg gatggctttc ttgccgccaa ggatctgatg gcgcagggga tcaagatctg 780
atcaagagac aggatgagga tcgtttcgca tggagaaaaa gatcacgggc tacactaccg 840
tggacatctc gcaatggcat cgcaaggaac acttcgaggc atttcaaagc gtggcacaat 900
gtacttacaa ccagaccgtc cagctggata ttaccgcgtt tttgaagacc gttaagaaaa 960
acaagcacaa gttttatcca gcctttatcc atattctcgc ccgcttgatg aatgcgcacc 1020
ccgaatttcg tatggccatg aaagatggtg agctcgttat ctgggactca gtccatccat 1080
gctataccgt tttccacgaa caaactgaaa ctttttcttc gctgtggtcc gaatatcacg 1140
atgatttccg ccaatttttg catatctaca gccaagatgt cgcgtgctat ggtgaaaacc 1200
tggcttactt tcccaaggga ttcatcgaga atatgttctt tgtttcagca aacccctggg 1260
tgtccttcac gtcgtttgac ttgaacgtgg ccaatatgga taatttcttc gctccagttt 1320
tcaccatggg taagtactat acccaaggag acaaggtcct tatgccactt gcaatccaag 1380
tacaccacgc agtctgcgat ggtttccatg tgggacgcat gcttaacgaa ctccaacagt 1440
actgtgatga atggcaaggc ggcgcgtagc cccccaaccg aagttgaggg gatttttgaa 1500
tcctcggtcc cccttgtcct tccagtcgaa gatagggact cttaacatcc caaaatcttt 1560
ctagaagatc tcctacaata ttctcagctg ccatggaaaa tcgatgttct tcttttattc 1620
tctcaagatt ttcaggctgt atattaaaac ttatattaag aactatgcta accacctcat 1680
caggaaccgt tgtaggtggc gtgggttttc ttggcaatcg actctcatga aaactacgag 1740
ctaaatattc aatatgttcc tcttgaccaa ctttattctg catttttttt gaacgaggtt 1800
tagagcaagc ttcaggaaac tgagacagga attttattaa aaatttaaat tttgaagaaa 1860
gttcagggtt aatagcatcc attttttgct ttgcaagttc ctcagcattc ttaacaaaag 1920
acgtctcttt tgacatgttt aaagtttaaa cctcctgtgt gaaattatta tccgctcata 1980
attccacaca ttatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 2040
agctaactca cattaattgc gttgcgctca ctgccaattg ctttccagtc gggaaacctg 2100
tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 2160
cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 2220
gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 2280
aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 2340
gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 2400
aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 2460
gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 2520
ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 2580
cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 2640
ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 2700
actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 2760
tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 2820
gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 2880
ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 2940
cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 3000
ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 3060
tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 3120
agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 3180
gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 3240
ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 3300
gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 3360
cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 3420
acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 3480
cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 3540
cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 3600
ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 3660
tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 3720
atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 3780
tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 3840
actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 3900
aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 3960
ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 4020
ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 4080
cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat 4140
aggcgtatca cgaggcc 4157
<210> 109
<211> 4159
<212> DNA
<213> 人工序列
<220>
<223> 合成地构建的质粒pJDI436
<400> 109
gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc 60
ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc 120
acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact 180
cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt 240
ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc 300
ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt 360
ttcagcaaga ttttgaggag tgttcagtct ccgtgaactc gacatacaca taaagtagct 420
tgcgtattta aaattatgaa cctaaggggt ttagcacttc acgctgccgc aagcactcag 480
ggcgcaaggg ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag aaacggtgct 540
gaccccggat gaatgtcagc tactgggcta tctggacaag ggaaaacgca agcgcaaaga 600
gaaagcaggt agcttgcagt gggcttacat ggcgatagct agactgggcg gttttatgga 660
cagcaagcga accggaattg ccagctgggg cgccctctgg taaggttggg aagccctgca 720
aagtaaactg gatggctttc ttgccgccaa ggatctgatg gcgcagggga tcaagatctg 780
atcaagagac aggatgagga tcgtttcgca tggagaaaaa gatcacgggc tacactaccg 840
tggacatctc gcaatggcat cgcaaggaac acttcgaggc atttcaaagc gtggcacaat 900
gtacttacaa ccagaccgtc cagctggata ttaccgcgtt tttgaagacc gttaagaaaa 960
acaagcacaa gttttatcca gcctttatcc atattctcgc ccgcttgatg aatgcgcacc 1020
ccgaatttcg tatggccatg aaagatggtg agctcgttat ctgggactca gtccatccat 1080
gctataccgt tttccacgaa caaactgaaa ctttttcttc gctgtggtcc gaatatcacg 1140
atgatttccg ccaatttttg catatctaca gccaagatgt cgcgtgctat ggtgaaaacc 1200
tggcttactt tcccaaggga ttcatcgaga atatgttctt tgtttcagca aacccctggg 1260
tgtccttcac gtcgtttgac ttgaacgtgg ccaatatgga taatttcttc gctccagttt 1320
tcaccatggg taagtactat acccaaggag acaaggtcct tatgccactt gcaatccaag 1380
tacaccacgc agtctgcgat ggtttccatg tgggacgcat gcttaacgaa ctccaacagt 1440
actgtgatga atggcaaggc ggcgcgtagc cccccaaccg aagttgaggg gatttttgaa 1500
tcctcggtcc cccttgtcct tccagtcgat tggggtatgg ggggggcggt caaaaaatct 1560
ttctagaaga tctcctacaa tattctcagc tgccatggaa aatcgatgtt cttcttttat 1620
tctctcaaga ttttcaggct gtatattaaa acttatatta agaactatgc taaccacctc 1680
atcaggaacc gttgtaggtg gcgtgggttt tcttggcaat cgactctcat gaaaactacg 1740
agctaaatat tcaatatgtt cctcttgacc aactttattc tgcatttttt ttgaacgagg 1800
tttagagcaa gcttcaggaa actgagacag gaattttatt aaaaatttaa attttgaaga 1860
aagttcaggg ttaatagcat ccattttttg ctttgcaagt tcctcagcat tcttaacaaa 1920
agacgtctct tttgacatgt ttaaagttta aacctcctgt gtgaaattat tatccgctca 1980
taattccaca cattatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 2040
tgagctaact cacattaatt gcgttgcgct cactgccaat tgctttccag tcgggaaacc 2100
tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 2160
ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 2220
cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 2280
gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 2340
tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 2400
agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 2460
tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 2520
cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 2580
ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 2640
ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 2700
ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 2760
ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 2820
cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 2880
gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 2940
atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 3000
ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 3060
gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 3120
tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 3180
ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 3240
taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 3300
gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 3360
gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 3420
ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 3480
aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 3540
gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 3600
cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 3660
actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 3720
caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 3780
gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 3840
ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 3900
caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 3960
tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 4020
gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 4080
cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa 4140
ataggcgtat cacgaggcc 4159
<210> 110
<211> 41
<212> RNA
<213> 人工序列
<220>
<223> 合成地构建的RNA靶向cTAG M
<400> 110
uaauuucuac ucuuguagau gguuaaagau gguuaaauga u 41
<210> 111
<211> 41
<212> RNA
<213> 人工序列
<220>
<223> 合成地构建的RNA靶向cTAG N
<400> 111
uaauuucuac ucuuguagau ggauguuaag agucccuauc u 41
<210> 112
<211> 41
<212> RNA
<213> 人工序列
<220>
<223> 合成地构建的RNA靶向cTAG O
<400> 112
uaauuucuac ucuuguagau accgcccccc ccauacccca a 41
<210> 113
<211> 43
<212> RNA
<213> 人工序列
<220>
<223> 合成地构建的RNA靶向cTAG P
<400> 113
uaauuucuac ucuuguagau aggaguguuc agucuccgug aac 43
<210> 114
<211> 9
<212> PRT
<213> 未知
<220>
<223> P53 TAD1转录因子结构域
<400> 114
Glu Thr Phe Ser Asp Leu Trp Lys Leu
1 5
<210> 115
<211> 9
<212> PRT
<213> 未知
<220>
<223> P53 TAD2转录因子结构域
<400> 115
Asp Asp Ile Glu Gln Trp Phe Thr Glu
1 5
<210> 116
<211> 8
<212> PRT
<213> 未知
<220>
<223> MLL转录因子结构域
<400> 116
Asp Ile Met Asp Phe Val Leu Lys
1 5
<210> 117
<211> 9
<212> PRT
<213> 未知
<220>
<223> EA2转录因子结构域
<400> 117
Asp Leu Leu Asp Phe Ser Met Met Phe
1 5
<210> 118
<211> 9
<212> PRT
<213> 未知
<220>
<223> Rtg3转录因子结构域
<400> 118
Glu Thr Leu Asp Phe Ser Leu Val Thr
1 5
<210> 119
<211> 9
<212> PRT
<213> 未知
<220>
<223> CREB转录因子结构域
<400> 119
Arg Lys Ile Leu Asn Asp Leu Ser Ser
1 5
<210> 120
<211> 9
<212> PRT
<213> 未知
<220>
<223> CREBaB6转录因子结构域
<400> 120
Glu Ala Ile Leu Ala Glu Leu Lys Lys
1 5
<210> 121
<211> 9
<212> PRT
<213> 未知
<220>
<223> Gli3转录因子结构域
<400> 121
Asp Asp Val Val Gln Tyr Leu Asn Ser
1 5
<210> 122
<211> 9
<212> PRT
<213> 未知
<220>
<223> Gal4转录因子结构域
<400> 122
Asp Asp Val Tyr Asn Tyr Leu Phe Asp
1 5
<210> 123
<211> 9
<212> PRT
<213> 未知
<220>
<223> Oaf1转录因子结构域
<400> 123
Asp Leu Phe Asp Tyr Asp Phe Leu Val
1 5
<210> 124
<211> 9
<212> PRT
<213> 未知
<220>
<223> Pip2转录因子结构域
<400> 124
Asp Phe Phe Asp Tyr Asp Leu Leu Phe
1 5
<210> 125
<211> 9
<212> PRT
<213> 未知
<220>
<223> Pdr1转录因子结构域
<400> 125
Glu Asp Leu Tyr Ser Ile Leu Trp Ser
1 5
<210> 126
<211> 9
<212> PRT
<213> 未知
<220>
<223> Pdr3转录因子结构域
<400> 126
Thr Asp Leu Tyr His Thr Leu Trp Asn
1 5

Claims (112)

1.一种用于调节宿主细胞基因的表达的重组模块化CRISPR DNA构建体，所述构建体包括CRISPR多克隆位点，所述多克隆位点包括：
a)至少两个不同的克隆标签(cTAG)，其中每个cTAG包括：
i)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序PAM可操作地连接的原间隔子序列；其中所述经过验证的CRISPR着陆位点中的至少一个在所述模块化CRISPR DNA构建体中是独特的；以及
b)一或多个DNA插入部分；
i)其中所述不同的cTAG中的每个cTAG围绕所述一或多个DNA插入部分中的每个DNA插入部分分布在侧翼位置；
其中所述构建体进一步包括：
c)第一核酸，所述第一核酸对催化灭活CRISPR酶进行编码；
d)第二核酸，所述第二核酸对能够将(c)的所述催化灭活CRISPR酶募集到DNA靶位点的引导RNA进行编码。
2.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体包括第一复制起点。
3.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体包括多于一个复制起点。
4.根据权利要求2所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPR DNA构建体包括第一复制起点和第二复制起点。
5.根据权利要求2所述的重组模块化CRISPR DNA构建体，其中所述第一复制起点能够维持大肠杆菌中的质粒。
6.根据权利要求4或5所述的重组模块化CRISPR DNA构建体，其中所述第二复制起点能够维持谷氨酸棒状杆菌中的质粒。
7.根据权利要求2所述的重组模块化CRISPR DNA构建体，其中所述第一复制起点能够维持大肠杆菌中的质粒，第二复制起点能够维持酿酒酵母中的质粒，并且第三复制起点能够维持谷氨酸棒状杆菌中的质粒。
8.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPRDNA构建体包括针对可选择标志物进行编码的插入部分。
9.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中至少一个复制起点包括在所述CRISPR多克隆位点内的插入部分内。
10.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述第一核酸包括在所述CRISPR多克隆位点内的插入部分内。
11.根据权利要求10所述的重组模块化CRISPR DNA构建体，其中包括所述第一核酸的所述插入部分进一步包括可选择标志物。
12.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述第二核酸包括在所述CRISPR多克隆位点内的插入部分内。
13.根据权利要求12所述的重组模块化CRISPR DNA构建体，其中包括所述第二核酸的所述插入部分进一步包括可选择标志物。
14.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述第一核酸和所述第二核酸各自包括在所述CRISPR多克隆位点内的其自身的插入部分内。
15.根据权利要求14所述的重组模块化CRISPR DNA构建体，其中包括所述第一核酸和所述第二核酸的所述插入部分各自包括可选择标志物。
16.根据权利要求15所述的重组模块化CRISPR DNA构建体，其中包括所述第一核酸的所述插入部分中包括的所述可选择标志物和包括所述第二核酸的所述插入部分中包括的所述可选择标志物是不同的。
17.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述第一核酸和所述第二核酸包括在所述CRISPR多克隆位点内的同一插入部分内。
18.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述第一核酸与启动子可操作地连接。
19.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述第一核酸与终止子可操作地连接。
20.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述第二核酸与启动子可操作地连接。
21.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述第二核酸与终止子可操作地连接。
22.根据权利要求18和20中任一权利要求所述的重组模块化CRISPR DNA构建体，其中所述启动子是异源启动子。
23.根据权利要求18和20中任一权利要求所述的重组模块化CRISPR DNA构建体，其中所述启动子是组成型启动子。
24.根据权利要求18和20中任一权利要求所述的重组模块化CRISPR DNA构建体，其中所述启动子是诱导型启动子。
25.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述第一核酸对与转录激活蛋白翻译融合的催化灭活CRISPR酶进行编码。
26.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述第一核酸对与转录灭活蛋白翻译融合的催化灭活CRISPR酶进行编码。
27.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述第一核酸对与转录阻遏物翻译融合的催化灭活CRISPR酶进行编码。
28.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述构建体进一步包括(e)第三核酸，所述第三核酸对在被表达时能够将自身与所述催化灭活CRISPR酶连接的转录激活蛋白进行编码。
29.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述构建体进一步包括(e)第三核酸，所述第三核酸对在被表达时能够将自身与所述催化灭活CRISPR酶连接的转录灭活蛋白进行编码。
30.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述构建体进一步包括(e)第三核酸，所述第三核酸对在被表达时能够将自身与所述催化灭活CRISPR酶连接的转录阻遏蛋白进行编码。
31.根据权利要求28所述的重组模块化CRISPR DNA构建体，其中所述转录激活蛋白经由连接适配子或通过蛋白质-蛋白质相互作用将自身与所述催化灭活CRISPR酶连接。
32.根据权利要求29所述的重组模块化CRISPR DNA构建体，其中所述转录灭活蛋白经由连接适配子或通过蛋白质-蛋白质相互作用将自身与所述催化灭活CRISPR酶连接。
33.根据权利要求30所述的重组模块化CRISPR DNA构建体，其中所述转录阻遏蛋白经由连接适配子或通过蛋白质-蛋白质相互作用将自身与所述催化灭活CRISPR酶连接。
34.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述第二核酸对与能够将自身与转录激活蛋白连接的适配子可操作地连接的引导RNA进行编码。
35.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述第二核酸对与能够将自身与转录灭活蛋白连接的适配子可操作地连接的引导RNA进行编码。
36.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述第二核酸对与能够将自身与转录阻遏蛋白连接的适配子可操作地连接的引导RNA进行编码。
37.根据权利要求25、28、31和34中任一权利要求所述的重组模块化CRISPR DNA构建体，其中所述转录激活蛋白选自由以下组成的群组：VP16、VP64和VP160、VPR。
38.根据权利要求26、29、32和35和16.1中任一权利要求所述的重组模块化CRISPRDNA构建体，其中所述转录灭活蛋白选自由以下组成的群组：Mxi1、Tbx3、KRAB、EnR和SID。
39.根据权利要求27、30、33和36中任一权利要求所述的重组模块化CRISPR DNA构建体，其中所述转录阻遏蛋白选自由以下组成的群组：Mxi1、Tbx3、KRAB、EnR和SID。
40.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPRDNA构建体是环状的。
41.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPRDNA构建体是线性的。
42.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPRDNA构建体被整合到生物体的基因组中。
43.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述不同的cTAG中的至少一个包括至少两个经过验证的CRISPR着陆位点。
44.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述CRISPR着陆位点中的至少一个是用于Cas9核酸内切酶。
45.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述CRISPR着陆位点中的至少一个是用于Cpf1核酸内切酶。
46.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述不同的cTAG中的至少一个包括罕见的(≥8个碱基长)限制性核酸内切酶位点。
47.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中每个cTAG包括罕见的(≥8个碱基长)限制性核酸内切酶位点。
48.根据权利要求1所述的模块化CRISPR DNA构建体，其中所述催化灭活CRISPR酶是突变的Cas9核酸内切酶。
49.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述催化灭活CRISPR酶是突变的Cpf1核酸内切酶。
50.根据权利要求1所述的模块化CRISPR DNA构建体，其中所述催化灭活CRISPR酶选自表1的载体中包含的dCas9基因中。
51.根据权利要求1所述的模块化CRISPR DNA构建体，其中所述催化灭活CRISPR酶选自由以下组成的群组：新凶手弗朗西丝菌(UniProtKB—A0Q7Q2(CPF1_FRATN))、毛螺菌科细菌(UniProtKB—A0A182DWE3(A0A182DWE3_9FIRM))和氨基酸球菌(UniProtKB—U2UMQ6(CPF1ACISB)。
52.根据权利要求1所述的模块化CRISPR DNA构建体，其中所述催化灭活CRISPR酶是AsCpf1(D908A)。
53.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述重组模块化CRISPRDNA构建体针对能够将(c)的所述催化灭活CRISPR酶募集到DNA靶位点的多于一个引导RNA进行编码。
54.根据权利要求53所述的重组模块化CRISPR DNA构建体，其中所述引导RNA中的至少一个包括与所述构建体中编码的另一个引导RNA不同的序列。
55.根据权利要求53所述的重组模块化CRISPR DNA构建体，其中所述引导RNA中的至少一个靶向与所述构建体中编码的另一个引导RNA不同的DNA靶位点序列。
56.根据权利要求1所述的重组模块化CRISPR DNA构建体，其中所述重组模块化CRISPRDNA构建体针对多于一种催化灭活CRISPR酶进行编码。
57.根据权利要求56所述的重组模块化CRISPR DNA构建体，其中所述催化灭活CRISPR酶中的至少一种包括与所述构建体中编码的另一种催化灭活CRISPR酶不同的序列。
58.根据权利要求1所述的插入部分，其中所述cTAG中的一或多个选自由SEQ ID NO:65-74、78-81和其组合组成的群组。
59.一种宿主细胞，其包括根据权利要求1到57中任一权利要求所述的重组模块化CRISPR DNA构建体。
60.一种调节一或多个宿主细胞基因的表达的高通量方法，所述方法包括将根据权利要求1所述的重组模块化CRISPR DNA构建体引入宿主细胞的步骤；其中引导RNA的DNA靶位点位于宿主细胞基因组内。
61.根据权利要求60所述的高通量方法，其中将所述重组模块化CRISPR DNA构建体的至少一个插入部分整合到所述宿主细胞的基因组中。
62.根据权利要求60所述的高通量方法，其中所述重组模块化CRISPR DNA构建体作为染色体外DNA保留在所述宿主细胞中。
63.根据权利要求60所述的高通量方法，其中将根据权利要求18到24中任一权利要求所述的重组模块化CRISPR DNA构建体引入所述宿主细胞中。
64.根据权利要求63所述的高通量方法，其进一步包括使所述宿主细胞与能够增加诱导型启动子的表达的化合物接触的步骤。
65.一种用于筛选CRISPR酶变体的重组模块化CRISPR DNA构建体，所述构建体包括CRISPR多克隆位点，所述多克隆位点包括：
a)至少两个不同的克隆标签(cTAG)，其中每个cTAG包括：
i)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序PAM可操作地连接的原间隔子序列；其中所述经过验证的CRISPR着陆位点中的至少一个在所述模块化CRISPR DNA构建体中是独特的；以及
b)一或多个DNA插入部分；
i)其中所述不同的cTAG中的每个cTAG围绕所述一或多个DNA插入部分中的每个DNA插入部分分布在侧翼位置；
其中所述构建体进一步包括：
c)第一核酸，所述第一核酸对蛋白质进行编码；
d)第二核酸，所述第二核酸对能够与DNA靶位点结合的引导RNA进行编码。
66.一种用于筛选CRISPR酶变体的重组模块化CRISPR DNA构建体，所述构建体包括CRISPR多克隆位点，所述多克隆位点包括：
a)至少两个不同的克隆标签(cTAG)，其中每个cTAG包括：
i)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序PAM可操作地连接的原间隔子序列；其中所述经过验证的CRISPR着陆位点中的至少一个在所述模块化CRISPR DNA构建体中是独特的；以及
b)一或多个DNA插入部分；
i)其中所述不同的cTAG中的每个cTAG围绕所述一或多个DNA插入部分中的每个DNA插入部分分布在侧翼位置；
其中所述构建体进一步包括：
c)第一核酸，所述第一核酸对CRISPR酶或疑似具有CRISPR功能的酶(“推定的CRISPR酶”)进行编码；
d)第二核酸，所述第二核酸对能够与DNA靶位点结合的引导RNA进行编码。
67.一种宿主细胞，其包括根据权利要求66所述的重组模块化CRISPR DNA构建体。
68.一种筛选CRISPR酶变体的高通量方法，所述方法包括以下步骤：
a)将根据权利要求66所述的重组模块化CRISPR DNA构建体引入宿主细胞中；
其中引导RNA的DNA靶位点位于宿主细胞基因组内；以及
b)测量在所述DNA靶位点处发生的DNA切割的程度。
69.一种用于调节宿主细胞基因的表达或工程化宿主细胞的基因组的重组模块化CRISPR DNA构建体，所述构建体包括CRISPR多克隆位点，所述多克隆位点包括：
a)至少两个不同的克隆标签(cTAG)，其中每个cTAG包括：
i)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序PAM可操作地连接的原间隔子序列；其中所述经过验证的CRISPR着陆位点中的至少一个在所述模块化CRISPR DNA构建体中是独特的；以及
b)一或多个DNA插入部分；
i)其中所述不同的cTAG中的每个cTAG围绕所述一或多个DNA插入部分中的每个DNA插入部分分布在侧翼位置；以及
其中所述一或多个DNA插入部分包括针对CRISPR功能调节剂进行编码的DNA。
70.根据权利要求69所述的重组模块化CRISPR DNA构建体，其中包括针对CRISPR功能调节剂进行编码的所述DNA的所述插入部分进一步包括可选择标志物。
71.根据权利要求70所述的重组模块化CRISPR DNA构建体，其中所述CRISPR功能调节剂选自由以下组成的群组：复制起点、可选择标志物、反向可选择标志物、抗CRISPR蛋白、启动子、终止子、dCas9蛋白、dCpf1蛋白、条形码、Cas9蛋白、Cpf1蛋白、DNA供体和促进多重化基因组编辑的蛋白质。
72.一种宿主细胞，其包括根据权利要求69到71中任一权利要求所述的重组模块化CRISPR DNA构建体。
73.根据权利要求72所述的宿主细胞，其中所述宿主细胞包括对催化活性CRISPR酶进行编码的核酸分子和能够将所述催化活性CRISPR酶募集到DNA靶位点的引导RNA。
74.根据权利要求72所述的宿主细胞，其中所述宿主细胞包括对催化灭活CRISPR酶进行编码的核酸分子和能够将所述催化灭活CRISPR酶募集到DNA靶位点的引导RNA。
75.根据权利要求74所述的宿主细胞，其中所述催化灭活CRISPR酶与转录激活蛋白翻译融合。
76.根据权利要求74所述的宿主细胞，其中所述催化灭活CRISPR酶与转录灭活蛋白翻译融合。
77.根据权利要求74所述的宿主细胞，其中所述催化灭活CRISPR酶与转录阻遏蛋白翻译融合。
78.根据权利要求74所述的宿主细胞，其中所述宿主细胞进一步包括对转录激活蛋白进行编码的核酸分子，所述转录激活蛋白在表达时能够将自身与所述催化灭活CRISPR酶连接。
79.根据权利要求74所述的宿主细胞，其中所述宿主细胞进一步包括对转录灭活蛋白进行编码的核酸分子，所述转录灭活蛋白在表达时能够将自身与所述催化灭活CRISPR酶连接。
80.根据权利要求74所述的宿主细胞，其中所述宿主细胞进一步包括对转录阻遏蛋白进行编码的核酸分子，所述转录阻遏蛋白在表达时能够将自身与所述催化灭活CRISPR酶连接。
81.根据权利要求78所述的宿主细胞，其中所述转录激活蛋白经由连接适配子或通过蛋白质-蛋白质相互作用将自身与所述催化灭活CRISPR酶连接。
82.根据权利要求79所述的宿主细胞，其中所述转录灭活蛋白经由连接适配子或通过蛋白质-蛋白质相互作用将自身与所述催化灭活CRISPR酶连接。
83.根据权利要求80所述的宿主细胞，其中所述转录阻遏蛋白经由连接适配子或通过蛋白质-蛋白质相互作用将自身与所述催化灭活CRISPR酶连接。
84.根据权利要求81所述的宿主细胞，其中所述引导RNA与能够将自身与转录激活蛋白连接的适配子可操作地连接。
85.根据权利要求82所述的宿主细胞，其中所述引导RNA与能够将自身与转录灭活蛋白连接的适配子可操作地连接。
86.根据权利要求83所述的宿主细胞，其中所述引导RNA与能够将自身与转录阻遏蛋白连接的适配子可操作地连接。
87.根据权利要求75、81和84中任一权利要求所述的宿主细胞，其中所述转录激活蛋白选自由以下组成的群组：VP16、VP64和VP160、VPR。
88.根据权利要求76、82和85中任一权利要求所述的宿主细胞，其中所述转录灭活蛋白选自由以下组成的群组：Mxi1、Tbx3、KRAB、EnR和SID。
89.根据权利要求77、83和86中任一权利要求所述的宿主细胞，其中所述转录激活蛋白选自由以下组成的群组：Mxi1、Tbx3、KRAB、EnR和SID。
90.根据权利要求69所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPRDNA构建体是环状的。
91.根据权利要求69所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPRDNA构建体是线性的。
92.根据权利要求69所述的重组模块化CRISPR DNA构建体，其中所述模块化CRISPRDNA构建体被整合到生物体的基因组中。
93.根据权利要求69所述的重组模块化CRISPR DNA构建体，其中所述不同的cTAG中的至少一个包括至少两个经过验证的CRISPR着陆位点。
94.根据权利要求69所述的重组模块化CRISPR DNA构建体，其中所述CRISPR着陆位点中的至少一个是用于Cas9核酸内切酶。
95.根据权利要求69所述的重组模块化CRISPR DNA构建体，其中所述CRISPR着陆位点中的至少一个是用于Cpf1核酸内切酶。
96.根据权利要求69所述的重组模块化CRISPR DNA构建体，其中所述不同的cTAG中的至少一个包括罕见的(≥8个碱基长)限制性核酸内切酶位点。
97.根据权利要求73所述的宿主细胞，其中所述催化灭活CRISPR酶是突变的Cas9核酸内切酶。
98.根据权利要求73所述的宿主细胞，其中所述催化灭活CRISPR酶是突变的Cpf1核酸内切酶。
99.根据权利要求73所述的宿主细胞，其中所述宿主细胞包括多于一个引导RNA。
100.根据权利要求99所述的宿主细胞，其中所述引导RNA中的至少一个包括与另一个引导RNA不同的序列。
101.根据权利要求99所述的宿主细胞，其中所述引导RNA中的至少一个靶向与另一个引导RNA不同的DNA靶位点序列。
102.根据权利要求74所述的宿主细胞，其中所述宿主细胞包括多于一种催化灭活CRISPR酶。
103.根据权利要求102所述的宿主细胞，其中所述催化灭活CRISPR酶中的至少一种包括与所述构建体中编码的另一种催化灭活CRISPR酶不同的序列。
104.根据权利要求66所述的插入部分，其中所述cTAG中的一或多个选自由SEQ ID NO:65-74、78-81和其组合组成的群组。
105.一种调节一或多个宿主细胞基因的表达的高通量方法，所述方法包括将根据权利要求66所述的重组模块化CRISPR DNA构建体引入宿主细胞中的步骤；其中引导RNA的DNA靶位点位于宿主细胞基因组内。
106.根据权利要求105所述的高通量方法，其中将所述重组模块化CRISPR DNA构建体的至少一个插入部分整合到所述宿主细胞的基因组中。
107.根据权利要求105或106所述的高通量方法，其中所述插入部分调节CRISPR蛋白的功能。
108.根据权利要求105或106所述的高通量方法，其中所述插入部分调节引导RNA的表达。
109.根据权利要求105或106所述的高通量方法，其中所述重组模块化CRISPR DNA构建体作为染色体外DNA保留在所述宿主细胞中。
110.一种用于筛选CRISPR酶变体的重组模块化CRISPR DNA构建体，所述构建体包括CRISPR多克隆位点，所述多克隆位点包括：
a)至少两个不同的克隆标签(cTAG)，其中每个cTAG包括：
i)一或多个经过验证的CRISPR着陆位点，每个经过验证的CRISPR着陆位点包括与原间隔子邻近基序PAM可操作地连接的原间隔子序列；其中所述经过验证的CRISPR着陆位点中的至少一个在所述模块化CRISPR DNA构建体中是独特的；以及
b)一或多个DNA插入部分；
i)其中所述不同的cTAG中的每个cTAG围绕所述一或多个DNA插入部分中的每个DNA插入部分分布在侧翼位置；
其中所述构建体进一步包括：
c)第一核酸，所述第一核酸对CRISPR酶或疑似具有CRISPR功能的酶(“推定的CRISPR酶”)进行编码；以及
d)第二核酸，所述第二核酸对能够与DNA靶位点结合的引导RNA进行编码。
111.一种在宿主细胞中筛选CRISPR活性的高通量方法，所述方法包括以下步骤：
a)将根据权利要求65或66所述的重组模块化CRISPR DNA构建体引入宿主细胞中；其中引导RNA的DNA靶位点位于宿主细胞基因组内；以及
b)测量在所述DNA靶位点处发生的DNA切割的程度。
112.一种在宿主细胞中筛选CRISPRi活性的高通量方法，所述方法包括以下步骤：
a)将根据权利要求65或66所述的重组模块化CRISPR DNA构建体引入宿主细胞中；其中引导RNA的DNA靶位点位于宿主细胞基因组内；以及
b)测量在所述DNA靶位点处发生的转录调节的程度。
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