CN114990082A - 一种突变体及其构建方法和应用 - Google Patents
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Abstract
本发明提供了一种突变体,其氨基酸序列为SEQ ID NO.1上发生下列位点的突变:Q332K;Q332K、G34D;Q332K、S38R;Q332K、Y128R;Q332K、G34D、S38R;Q332K、G34D、Y128R;Q332K、S38R、Y128R;Q332K、G34D、S38R、Y128R。本发明提供的精氨酸氧化酶突变体与野生型精氨酸氧化酶相比,突变体热稳定性更好。通过本发明所提供的构建方法得到的精氨酸氧化酶突变体热稳定性更好,在较高的温度下氧化L‑精氨酸时,仍然表现出优良的催化活性,在L‑精氨酸生物传感器检测及生物化工等领域具有较高的应用潜力。
Description
本申请是申请日为2020年12月30日、申请号为202011624067.4、发明名称为“一种突变体及其构建方法和应用”的中国专利申请的分案申请。
技术领域
本发明涉及生物技术领域,具体而言,涉及一种突变体及其构建方法和应用。
背景技术
L-氨基酸氧化酶是一类生物体内参与氨基酸氧化代谢的重要氧化还原酶,能够以氧分子为电子受体催化L-氨基酸氧化脱氨,生成相应的酮酸、氨(NH3)和过氧化氢(H2O2)。有些L-氨基酸氧化酶能够专一性识别特定氨基酸,而不受其他种类氨基酸的干扰,因而在手性胺类化合物拆分、α-酮酸生物合成、临床样本、食品及氨基酸发酵过程中氨基酸含量检测等领域都发挥着重要作用。其中,L-精氨酸氧化酶能够特异性识别L-精氨酸,是生命活动的重要催化剂,且催化反应条件温和、产物单一、耗能小、产物易分离,在食品、化工、环保、能源以及医药领域都有着广泛地应用。然而,由于天然L-精氨酸氧化酶在高温、极端的酸碱度、有机溶剂、非天然底物、产物抑制等苛刻条件下的稳定性及催化活性将大大降低,限制了其在工业生产方面的应用。
蛋白质工程是以蛋白质分子的结构规律及其生物功能的关系作为基础,通过化学、物理和分子生物学的手段进行基因修饰或基因合成,对现有蛋白质进行改造或制造一种新的蛋白质以满足人类对生产和生活的需求。理性设计是蛋白质工程改造中最常用的方法,利用计算机辅助分子模型结合定点突变,以实现蛋白质功能优化,如提高催化活性、热稳定性、耐酸碱性等。为了有效优化蛋白质的热稳定性,Markus Wyss等人于2001年提出Consensus Concept理论。和常规蛋白质理性设计方法基于蛋白质精确结构-功能关系不同的是,Consensus Concept理论是基于同源蛋白的氨基酸序列信息,从进化的角度分析能提高酶热稳定性的信息。本发明以Consensus Concept理论为指导思想,对L-氨基酸氧化酶家族序列进行整合分析,并结合生物信息学辅助,获得具高稳定性的新型L-精氨酸氧化酶ARod突变体。相关研究目前未见报道。
发明内容
因此,本发明的目的是针对现有的精氨酸氧化酶突变体的热稳定性不足的问题,提供一种具有热稳定性的突变体、所述突变体的基因序列以及包含该突变体的可溶性蛋白、固定化酶、重组工程细胞、重组载体,用于催化氧化L-精氨酸。
根据本申请公开的第一方面,提供了一种突变体,其氨基酸序列为SEQ ID NO.1上发生下列位点的突变:
Q332K;
Q332K、G34D;
Q332K、S38R;
Q332K、Y128R;
Q332K、G34D、S38R;
Q332K、G34D、Y128R;
Q332K、S38R、Y128R;
Q332K、G34D、S38R、Y128R。
在一些可能的实现方式中,所述突变体对应的氨基酸序列分别为SEQ ID NO.5、SEQ ID NO.8、SEQ ID NO.10、SEQ ID NO.11、SEQ ID NO.13、SEQ ID NO.14、SEQ ID NO.15、SEQ ID NO.16。
根据本申请公开的第二方面,所述基因序列被配置为SEQ ID NO.20、SEQ IDNO.23、SEQ ID NO.25、SEQ ID NO.26、SEQ ID NO.28、SEQ ID NO.29、SEQ ID NO.30、SEQ IDNO.31中的任一种,其中:
编码突变位点为Q332K的突变体的核酸序列为SEQ ID NO.20;
编码突变位点为G34D和Q332K的突变体的核酸序列为SEQ ID NO.23;
编码突变位点为S38R和Q332K的突变体的核酸序列为SEQ ID NO.25;
编码突变位点为Y128R和Q332K的突变体的核酸序列为SEQ ID NO.26;
编码突变位点为G34D、S38R和Q332K的突变体的精氨酸氧化酶突变体的核酸序列为SEQ ID NO.28;
编码突变位点为S38R、Y128R和Q332K的突变体的核酸序列为SEQ ID NO.29;
编码突变位点为G34D、Y128R和Q332K的突变体的核酸序列为SEQ ID NO.30;
编码突变位点为G34D、S38R、Y128R和Q332K的突变体的核酸序列为SEQ ID NO.31。
进一步地,所述突变位点G34D的扩增引物对的核酸序列为SEQ ID NO.32和SEQ IDNO.33。
进一步地,所述突变位点S38R的扩增引物对的核酸序列为SEQ ID NO.34和SEQ IDNO.35。
进一步地,所述突变位点Y128R的扩增引物对的核酸序列为SEQ ID NO.36和SEQID NO.37。
进一步地,所述突变位点Q332K的扩增引物对的核酸序列为SEQ ID NO.38和SEQID NO.39。
根据本申请公开的第三方面,提供了一种可溶性蛋白,其被配置为包括如前述突变体。
根据本申请公开的第四方面,提供了一种固定化酶,其被配置为包括如前述突变体。
根据本申请公开的第五方面,提供了一种重组工程细胞,其被配置为包括如前述突变体的编码序列。
根据本申请公开的第六方面,提供了一种重组载体,其被配置为包括如前述突变体的编码序列。
根据本申请公开的第七方面,提供了前述突变体、可溶性蛋白、固定化酶、重组工程细胞或重组载体在催化氧化L-精氨酸中的应用。
本申请公开提供的突变体包括单点突变体和组合突变体,与野生型精氨酸氧化酶相比,其单点突变体和组合突变体在42℃下半衰期均更长;尤其是组合突变体,表现出单点突变体热稳定性的叠加效果,其半衰期大约是野生型精氨酸氧化酶的6倍。因此,本申请公开所提供的突变体的热稳定性更好,适于在较高的温度下催化氧化L-精氨酸。
本申请公开提供的突变体的构建方法,与理性设计基于蛋白质精确结构-功能关系不同的是,本发明以Consensus Concept理论为指导思想,从进化的角度分析能提高酶热稳定性的信息,对氨基酸氧化酶家族序列进行整合分析,并结合生物信息学及晶体学方法辅助,获得具高稳定性的新型精氨酸氧化酶突变体。
附图说明
为了更清楚地说明本发明实施方式的技术方案,下面将对实施方式中所需要使用的附图作简单地介绍,应当理解,以下附图仅示出了本发明的某些实施例,因此不应被看作是对范围的限定,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他相关的附图。
图1示出本申请公开的一个实施例的天然精氨酸氧化酶分子结构,图中箭头所指即为突变位点。
具体实施方式
为使本发明实施方式的目的、技术方案和优点更加清楚,下面将结合本发明实施方式中的附图,对本发明实施方式中的技术方案进行清楚、完整地描述,显然,所描述的实施方式是本发明一部分实施方式,而不是全部的实施方式。因此,以下对在附图中提供的本发明的实施方式的详细描述并非旨在限制要求保护的本发明的范围,而是仅仅表示本发明的选定实施方式。基于本发明中的实施方式,本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其他实施方式,都属于本发明保护的范围。
本申请公开所提供的精氨酸氧化酶的突变体中,天然的精氨酸氧化酶为来源于Oceanobacter kriegii的野生型精氨酸氧化酶,其氨基酸序列为SEQ ID NO.1。
在一些实施例中,该突变体的氨基酸序列为如下(a)或(b):
(a)至少取代、缺失或添加天然的精氨酸氧化酶的氨基酸序列SEQ ID NO.1的一个氨基酸,且所述突变体的氨基酸序列与SEQ ID NO.1具有90%以上同源性;或
(b)至少取代、缺失或添加天然的精氨酸氧化酶的氨基酸序列SEQ ID NO.1的一个氨基酸,且所述突变体的氨基酸序列与SEQ ID NO.1具有相同的功能。
在一些实施例中,突变体的氨基酸序列为SEQ ID NO.2~16中的任一种。具体地,在SEQ ID NO.1所示的氨基酸序列选择某一个位点进行单点突变,分别得到5种精氨酸氧化酶单点突变体,其突变位点为:G34D、S38R、Y128R、S195R、Q332K,将该5种精氨酸氧化酶单点突变体进行性质测定,筛选出4种热稳定性提高的精氨酸氧化酶突变体,其突变位点为:G34D、S38R、Y128R、Q332K,其氨基酸序列分别为SEQ ID NO.2、SEQ ID NO.3、SEQ ID NO.4、SEQ ID NO.5以及,
在SEQ ID NO.1所示的氨基酸序列选择多个突变位点进行组合:
1)如从上述4个突变位点中选择2个突变位点进行组合,分别得到以下6种热稳定性提高的精氨酸氧化酶突变体,其组合突变位点为:G34D/S38R、G34D/Y128R、G34D/Q332K、S38R/Y128R、S38R/Q332K、Y128R/Q332K,其氨基酸序列分别为SEQ ID NO.6、SEQ ID NO.7、SEQ ID NO.8、SEQ ID NO.9、SEQ ID NO.10、SEQ ID NO.11。
2)如从上述4个突变位点中选择3个突变位点进行组合,分别得到4种热稳定性提高的精氨酸氧化酶突变体,其组合突变位点为:
G34D/S38R/Y128R,
G34D/S38R/Q332K,
S38R/Y128R/Q332K,
G34D/Y128R/Q332K,其氨基酸序列分别为SEQ ID NO.12、SEQ ID NO.13、SEQ IDNO.14、SEQ ID NO.15。
3)如从上述4个突变位点中选择4个突变位点进行组合,得到1种热稳定性提高的精氨酸氧化酶突变体,其组合突变位点为:G34D/S38R/Y128R/Q332K,其氨基酸质序列为SEQID NO.16。
本申请公开的实施例提供了编码前述突变体的基因序列,该基因序列与具有不同的突变位点的氨基酸序列的对应关系如下:
编码突变位点为G34D的突变体的核酸序列为SEQ ID NO.17;
编码突变位点为S38R的突变体的核酸序列为SEQ ID NO.18;
编码突变位点为Y128R的突变体的核酸序列为SEQ ID NO.19;
编码突变位点为Q332K的突变体的核酸序列为SEQ ID NO.20;
编码突变位点为G34D和S38R的突变体的核酸序列为SEQ ID NO.21;
编码突变位点为G34D和Y128R的突变体的核酸序列为SEQ ID NO.22;
编码突变位点为G34D和Q332K的突变体的核酸序列为SEQ ID NO.23;
编码突变位点为S38R和Y128R的突变体的核酸序列为SEQ ID NO.24;
编码突变位点为S38R和Q332K的突变体的核酸序列为SEQ ID NO.25;
编码突变位点为Y128R和Q332K的突变体的核酸序列为SEQ ID NO.26;
编码突变位点为G34D、S38R和Y128R的突变体的精氨酸氧化酶突变体的核酸序列为SEQ ID NO.27;
编码突变位点为G34D、S38R和Q332K的突变体的精氨酸氧化酶突变体的核酸序列为SEQ ID NO.28;
编码突变位点为S38R、Y128R和Q332K的突变体的核酸序列为SEQ ID NO.29;
编码突变位点为G34D、Y128R和Q332K的突变体的核酸序列为SEQ ID NO.30;
编码突变位点为G34D、S38R、Y128R和Q332K的突变体的核酸序列为SEQ ID NO.31。
进一步地,突变位点G34D的扩增引物对的核酸序列为SEQ ID NO.32和SEQ IDNO.33。
进一步地,突变位点S38R的扩增引物对的核酸序列为SEQ ID NO.34和SEQ IDNO.35。
进一步地,突变位点Y128R的扩增引物对的核酸序列为SEQ ID NO.36和SEQ IDNO.37。
进一步地,突变位点Q332K的扩增引物对的核酸序列为SEQ ID NO.38和SEQ IDNO.39。
本申请公开的实施例还提供了一种前述突变体的构建方法,该方法包括:
S1:精氨酸氧化酶基因的克隆
将天然野生型精氨酸氧化酶基因以大肠杆菌为宿主细胞进行密码子优化,得到优化的基因序列,其核酸序列为SEQ ID NO.25,该核酸序列所表达的氨基酸序列为SEQ IDNO.1。
以SEQ ID NO.1作为目的基因,采用如下扩增引物对扩增目的基因:
F:5’-ACTGCTCATATGATGGATTCTAATAACCAATCCCCTCAG-3’(其中下划线为限制性内切酶NdeI识别位点);
R:5’-TCAGCTCTCGAGCTGTTGCGCGTGGTTTATGCCTTCG-3’(其中下划线为限制性内切酶XhoI识别位点)。
扩增条件为:在95℃下扩增2min,然后在56℃下扩增20sec、在72℃下扩增90sec,共30个循环,最后在72℃下扩增10min。
待反应结束,采用1.5%琼脂糖凝胶电泳检测PCR扩增产物,得到1.0kb的条带,其长度符合预期结果。按照试剂盒标准操作,回收、纯化该目的片段,使用限制性核酸内切酶XhoI和NdeI对该目的片段以及pET28a质粒进行双酶切,然后采用T4 DNA连接酶进行连接,将得到的连接产物转化至大肠杆菌BL21(DE3)感受态细胞中,将转化细胞涂布于含有100μg/ml卡那霉素的LB平板上,提取阳性克隆质粒,进行测序,结果显示克隆的精氨酸氧化酶基因序列正确,且已正确接入pET28a质粒,得到含有精氨酸氧化酶基因序列的重组质粒pET28a。
其中,PCR扩增酶为KOD高保真聚合酶(东洋纺)。
S2:精氨酸氧化酶的表达和纯化
将甘油管中的工程菌(含有精氨酸氧化酶基因序列的重组质粒pET28a)按体积比1%接种到含100μg/mL Kan的4mL LB培养基试管中,在37℃、220rpm条件下培养12h;取菌液4mL转接至含100μg/mL Kan的1L LB培养基摇瓶中,在37℃、220rpm条件下培养2.5h,使OD600达到0.6~0.8,加入1mM IPTG诱导剂,在25℃、200rpm条件下诱导培养14h。将发酵后收获的大肠杆菌菌体悬液超声破碎,再经过一步Ni-NTA亲和层析处理,得到纯度>95%的精氨酸氧化酶,其氨基酸序列为SEQ ID NO.1。
S3:精氨酸氧化酶同源蛋白的多序列比对及Consensus分析
S301:进入Pfam数据库主页(http://pfam.xfam.org/),在SEQUENCE SEARCH工具中输入精氨酸氧化酶的氨基酸序列进行搜索,服务器将直接反馈该蛋白整个家族的氨基酸序列的比对结果,将各个突变位点的各类氨基酸丰度以柱状图形式显示,该网站也可以自动生成该蛋白家族的consensus sequence;
S302:在NCBI的protein数据库及Pfam数据库中输入SEQ ID NO.1所示的氨基酸序列,利用Blast工具,找出所有与精氨酸氧化酶的氨基酸序列一致性大于50%的蛋白质序列,删除其中的重复出现的相同序列,将剩余氨基酸序列整理成fasta.格式,输入Clustalx1.83软件进行多序列比对,比对结果以aln.、dnd.和fasta.格式输出,其中,dnd.文件为构建进化树文件,aln.和fasta.文件为不同形式的序列文件;
将上述fasta.文件上传到Consensus Maker v2.0.0
(https://www.hiv.lanl.gov/content/sequence/CONSENSUS/consensus.html)
服务器,根据需要修改设置参数后,该在线软件将生成可以后期编辑的consensussequence。
S303:将精氨酸氧化酶的氨基酸序列与该家族共有序列(consensus sequence)以及各位点氨基酸的丰度图进行对照比较。
S4:精氨酸氧化酶的三维结构的模拟及突变热点的选择
S401:通过swissmodel在线工具对获得精氨酸氧化酶(SEQ ID NO.1)的三维结构预测;
S402:用PyMOL观测精氨酸氧化酶(氨基酸序列SEQ ID NO.1)的晶体结构,根据结构信息复查上述待选突变位点及突变形式,筛选出最有可能提高精氨酸氧化酶热稳定性的突变体位点。筛选条件具体如下:
(1)判断某一位点为待选位点的标准为:
①该家族大多数蛋白在该位点处氨基酸丰度总体高度较高;
②该位点氨基酸保守;
③该位点出现频率较高的氨基酸与精氨酸氧化酶在该位点处的氨基酸有较大的理化性质差异,如氢键、电荷差异、极性强弱、空间位阻大小等。
经过上述两步筛选,此时共剩余25个差异位点,大多数位于精氨酸氧化酶分子的表面,如图1所示,箭头所指即为突变位点。
(3)根据精氨酸氧化酶晶体结构,逐个详细分析上述25种突变形式,筛选出可能提高精氨酸氧化酶热稳定性的突变体。
主要判断准则为:①突变应消除原有的不利于热稳定的作用力形式,如静电排斥作用、电荷聚集等;②突变不应破坏现有的利于热稳定的作用力形式和稳定的蛋白结构;③突变应引入新的利于热稳定的作用力形式,如氢键、盐桥、疏水相互作用等。
共设计单点突变体5个,其突变位点分别为:
G34D、S38R、Y128R、S195R、Q332K;
将该5种精氨酸氧化酶单点突变体进行活力测定,筛选出4种热稳定性提高的精氨酸氧化酶突变体,其突变位点为:G34D、S38R、Y128R、Q332K,所对应的单点突变体的氨基酸序列分别为SEQ ID NO.2、SEQ ID NO.3、SEQ ID NO.4、SEQ ID NO.5。
S5:突变体的构建、表达及纯化
S501:精氨酸氧化酶单点突变体的构建
以S1中的重组质粒(含有精氨酸氧化酶基因序列的重组质粒pET28a)为模板,以带有突变位点的一对互补的寡核苷酸为扩增引物,用KOD高保真酶(TakaRa公司)进行全质粒PCR扩增,获得具有特定突变位点的重组质粒;
所采用的扩增引物对为:
(1)突变位点G34D的上下游扩增引物的核酸序列分别如下:
F(SEQ ID NO.32):
5’-TCAGCTGTCGACATGGATTCTAATAACCAATCCCCTCAG-3’;
R(SEQ ID NO.33):
5’-ACTGCTGCTAGCCTGTTGCGCGTGGTTTATGCCTTCGTATG-3’;
(2)突变位点S38R的上下游扩增引物的核酸序列分别如下:
F(SEQ ID NO.34):
5’-TCAGCTGGATCCATGGATTCTAATAACCAATCCCCTCAG-3’;
R(SEQ ID NO.35):
5’-ACTGCTAGATCTCTGTTGCGCGTGGTTTATGCCTTCGTATG-3’;
(3)突变位点Y128R的上下游扩增引物的核酸序列分别如下:
F(SEQ ID NO.36):
5’-TCAGCTGGTACCATGGATTCTAATAACCAATCCCCTCAG-3’;
R(SEQ ID NO.37):
5’-ACTGCTAAGCTTCTGTTGCGCGTGGTTTATGCCTTCGTATG-3’;
(4)突变位点Q332K的上下游扩增引物的核酸序列分别如下:
F(SEQ ID NO.38):
5’-TCAGCTATCGATATGGATTCTAATAACCAATCCCCTCAG-3’;
R(SEQ ID NO.39):
5’-ACTGCTCATATGCTGTTGCGCGTGGTTTATGCCTTCGTATG-3’;
扩增条件为:在95℃下扩增2min,然后在56℃下扩增20sec、在72℃下扩增90sec,共30个循环,最后在72℃下扩增10min;胶回收PCR扩增产物,采用DpnI酶(Fermentas公司)在37℃条件下消化胶回收产物2h,降解初始模板;将消化产物转化至大肠杆菌BL21(DE3)感受态细胞中,涂布到含有100μg/mL卡那霉素LB琼脂平板上,37℃过夜培养,筛选阳性克隆,测序验证,得到含精氨酸氧化酶单点突变体的重组菌;
S502:精氨酸氧化酶组合突变体的构建
使用与单点突变体相似的构建方法,将稳定性提高的单点突变体累加组合,在SEQID NO.1所示的氨基酸序列选择多个突变位点进行组合,如从上述4个突变位点中选择2~4个突变位点进行组合,分别得到不同的精氨酸氧化酶组合突变体:
(1)选择2个突变位点进行组合,可构建6种热稳定性提高的精氨酸氧化酶突变体及精氨酸氧化酶组合突变体,其组合突变位点分别为:
G34D/S38R、G34D/Y128R、G34D/Q332K、S38R/Y128R、S38R/Q332K、Y128R/Q332K,
该6种热稳定性提高的精氨酸氧化酶组合突变体的氨基酸序列分别为SEQ IDNO.6、SEQ ID NO.7、SEQ ID NO.8、SEQ ID NO.9、SEQ ID NO.10、SEQ ID NO.11;
(2)选择3个突变位点进行组合,可构建4种热稳定性提高的精氨酸氧化酶组合突变体,其组合突变位点分别为:
G34D/S38R/Y128R、G34D/S38R/Q332K、S38R/Y128R/Q332K、G34D/Y128R/Q332K,
该4种热稳定性提高的精氨酸氧化酶组合突变体的氨基酸序列分别为SEQ IDNO.12、SEQ ID NO.13、SEQ ID NO.14、SEQ ID NO.15;
(3)选择4个突变位点进行组合,可构建1种热稳定性提高的精氨酸氧化酶组合突变体,其组合突变位点分别为:
G34D/S38R/Y128R/Q332K,
该1种热稳定性提高的精氨酸氧化酶组合突变体的氨基酸序列为SEQ ID NO.16。
实验——精氨酸氧化酶突变体的酶学性质表征
将天然野生型精氨酸氧化酶以及实施例提供的多种精氨酸氧化酶突变体进行热稳定性测试,按照常规精氨酸氧化酶活力测定方法,具体为:
在一定温度下下孵育酶液,在不同处理时间取样,测定精氨酸氧化酶或精氨酸氧化酶突变体残余活力百分比,以残余活力百分比的ln值对时间t(min)作图,直线的斜率为失活常数kinact,由t1/2=ln2/kinact得到该种野生型精氨酸氧化酶或精氨酸氧化酶突变体在该温度下的半衰期。
实验结果表明,以上多种精氨酸氧化酶突变体中,有4种单点突变体和11种组合突变体的热稳定性得到明显改善,如表1所示:
表1.野生型精氨酸氧化酶、单点突变体和组合突变体的酶学性质表征
由表1得知,本发明提供的精氨酸氧化酶突变体包括单点突变体和组合突变体,与野生型精氨酸氧化酶相比,其单点突变体和组合突变体在42℃下半衰期均更长;尤其是组合突变体,表现出单点突变体热稳定性的叠加效果,其半衰期大约是野生型精氨酸氧化酶的6倍。基于此,本发明所提供的精氨酸氧化酶突变体的热稳定性更好,适于在较高的温度下催化氧化L-精氨酸。
尽管已经通过优选实施例进一步详细说明和描述了本发明,但是本发明不限于所公开的示例,并且本领域技术人员可以在不脱离本发明的范围的情况下从其中得出其他变型。凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
序列表
序列表
<110>中国科学院苏州生物医学工程技术研究所
<160>39
<210>1
<211>603
<212>PRT
<213>人工
<400>1
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSGGEQSIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRYRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LQSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>2
<211>603
<212> PRT
<400>2
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSDGEQSIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRYRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LQSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>3
<211>603
<212>PRT
<213>人工
<400>3
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSGGEQRIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRYRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LQSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>4
<211>603
<212>PRT
<213>人工
<400>4
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSGGEQSIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRRRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LQSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>5
<211>603
<212>PRT
<213>人工
<400>5
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSGGEQSIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRYRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LKSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>6
<211>603
<212>PRT
<213>人工
<400>6
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSDGEQRIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRYRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LQSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>7
<211>603
<212>PRT
<213>人工
<400>7
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSDGEQSIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRRRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LQSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>8
<211>603
<212>PRT
<213>人工
<400>8
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSDGEQSIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRYRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LKSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>9
<211>603
<212>PRT
<213>人工
<400>9
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSGGEQRIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRRRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LQSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>10
<211>603
<212>PRT
<213>人工
<400>10
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSGGEQRIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRYRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LKSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>11
<211>603
<212>PRT
<213>人工
<400>11
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSGGEQSIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRRRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LKSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>12
<211>603
<212>PRT
<213>人工
<400>12
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSDGEQRIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRRRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LQSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>13
<211>603
<212>PRT
<213>人工
<400>13
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSDGEQRIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRYRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LKSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>14
<211>603
<212>PRT
<213>人工
<400>14
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSDGEQSIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRRRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LKSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>15
<211>603
<212>PRT
<213>人工
<400>15
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSGGEQRIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRRRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LKSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>16
<211>603
<212>PRT
<213>人工
<400>16
MDSNNQSPQV LDQAIIGGGV SGVYSAWRLQ QQSDGEQRIA LFEYSNRIGG RLYSRKIPGL 60
DNVVAELGGM RWIAEDHPMV NSLVHELKLA IKEFPMGSSL PLDPKVKDSP KAGTENNLFY 120
LRGQYFRRRD FAECPDRIPY NLGWSERGYG PEDMQVKVMN LICPGFADMS LAEQMAVEVN 180
GKPMWQYGFW NLLESVLSNE AYQFMKDAGG YEANVANANA VTQLPATEYK DNTQFLGLKQ 240
GFQALPLTLC DCFEKLGGAV HMDMRLAEIR IDPASDTRYT LIFQPTSTDD SGKTTDTDDA 300
CVEVQAKKVI LAMPRRSLEL IKSDYFEDEW LKSNIPSVLI QKAFKMFMAY ESPWWRSLGL 360
VYGRSVTDLP IRQTYYMGTE CDASDVSVST NSLLMASYND IGTVPYWKGL EAGEPFEGYT 420
PAGMTLAAGE RIVPNHEFQI SDAMVQAAQR QLEAVHNQKQ LPQPYSAVYQ EWGHEPYGGG 480
WHEWKAGFEL DKVMQKMRHP VEGEDIYIVG EAYSYDQGWV EGALVVAESM LEEFYCLDAP 540
SWLKVSDASH AFLPSLCNWV PQLLTEPYPV PENDSATDAN AAIRATLNEV TTFAYEGINH 600
AQQ 603
<210>17
<211>1809
<212>DNA
<213>人工
<400>17
atggattcta ataaccaatc ccctcaggtt ttagaccaag ctattatcgg tggcggagtc 60
tcaggggtat attcggcctg gcgtttgcag caacagagtg gtgacgaaca aagcagtgca 120
ctttttgagt actctaatcg cattggaggg cgactctatt cccggaaaat ccccggtcta 180
gataacgtgg ttgcggaact gggcggaatg agatggatag ctgaggacca tccaatggtc 240
aattcattag tacacgaatt gaagcttgcc attaaagagt tcccgatggg gtcgagtctc 300
cctctagatc ccaaggtgaa agacagccca aaggcaggta ctgaaaacaa tctgttttac 360
ttaaggggcc agtatttccg ttaccgcgat tatgcggagt gtccggaccg aatcccttat 420
aacttgggat ggtctgaacg ggggtacggt cccgaggata tgcaagttaa agtcatgaat 480
cttatatgcc caggcttcgc tgacatgtcc ctcgccgaac agatggcagt agaggtgaac 540
ggaaagccga tgtggcaata tgggttttgg aatctactgg aatcagtttt atcgaacgag 600
gcgtaccagt tcatgaaaga tgctggtggc tatgaagcca atgtcgcaaa cgcgaatgct 660
gtaacccaat tgcctgccac agagtacaag gacaacacgc agtttcttgg actcaaacaa 720
gggttccagg cactacccct gactttatgt gattgctttg aaaagttggg tggcgcggtg 780
catatggaca tgagacttgc tgagattagg atcgatccag ccagtgacac ccgttataca 840
ctcatattcc aaccgacgag cactgatgac tctggaaaaa ccacagatac ggacgatgca 900
tgtgttgaag tccaggcgaa gaaagtaatt ctagctatgc ctcgccgatc cctggagtta 960
atcaagtcag actactttga agatgagtgg ttgcaatcga atatacccag tgtgcagatt 1020
cagaaagcct tcaagatgtt tatggcatat gaaagcccat ggtggcggtc tctcgggcta 1080
gtttacggta gatccgtcac tgacctgccg atcaggcaaa cctattacat gggcacagag 1140
tgcgatgcgt cagacgtatc ggtgagtacg aacagcttat tgatggcttc ttataatgat 1200
ataggaactg ttccttactg gaaagggctt gaagccggtg agcccttcga aggctatacc 1260
ccagcaggaa tgacactcgc ggctggggag cgtattgtcc cgaaccacga atttcagatc 1320
tccgacgcca tggtacaagc agcgcagcgc caactagagg ctgtgcataa tcagaagcaa 1380
ctgcctcagc cctactcagc cgtttatcaa gaatggggtc acgagccata cggcggaggg 1440
tggcatgaat ggaaagcagg tttcgagtta gataaggtca tgcagaaaat gcgacacccg 1500
gtagaaggcg aggacatata tattgtggga gaagcgtact cgtatgatca agggtgggtt 1560
gagggtgctt tggtcgtagc cgaaagtatg cttgaggaat tttactgtct cgacgcacct 1620
agctggctaa aggtgtctga tgcgtcccat gctttcctgc cctcattatg caactgggtt 1680
ccacagttgc ttacggagcc gtatcctgtc cccgaaaatg actcggccac tgatgcaaac 1740
gcggctatcc gggccaccct caatgaggta acaacgtttg catacgaagg cataaaccac 1800
gcgcaacag 1809
<210>18
<211>1809
<212>DNA
<213>人工
<400>18
atggattcta ataaccaatc ccctcaggtt ttagaccaag ctattatcgg tggcggagtc 60
tcaggggtat attcggcctg gcgtttgcag caacagagtg gtggcgaaca aagcagagca 120
ctttttgagt actctaatcg cattggaggg cgactctatt cccggaaaat ccccggtcta 180
gataacgtgg ttgcggaact gggcggaatg agatggatag ctgaggacca tccaatggtc 240
aattcattag tacacgaatt gaagcttgcc attaaagagt tcccgatggg gtcgagtctc 300
cctctagatc ccaaggtgaa agacagccca aaggcaggta ctgaaaacaa tctgttttac 360
ttaaggggcc agtatttccg ttaccgcgat tatgcggagt gtccggaccg aatcccttat 420
aacttgggat ggtctgaacg ggggtacggt cccgaggata tgcaagttaa agtcatgaat 480
cttatatgcc caggcttcgc tgacatgtcc ctcgccgaac agatggcagt agaggtgaac 540
ggaaagccga tgtggcaata tgggttttgg aatctactgg aatcagtttt atcgaacgag 600
gcgtaccagt tcatgaaaga tgctggtggc tatgaagcca atgtcgcaaa cgcgaatgct 660
gtaacccaat tgcctgccac agagtacaag gacaacacgc agtttcttgg actcaaacaa 720
gggttccagg cactacccct gactttatgt gattgctttg aaaagttggg tggcgcggtg 780
catatggaca tgagacttgc tgagattagg atcgatccag ccagtgacac ccgttataca 840
ctcatattcc aaccgacgag cactgatgac tctggaaaaa ccacagatac ggacgatgca 900
tgtgttgaag tccaggcgaa gaaagtaatt ctagctatgc ctcgccgatc cctggagtta 960
atcaagtcag actactttga agatgagtgg ttgcaatcga atatacccag tgtgcagatt 1020
cagaaagcct tcaagatgtt tatggcatat gaaagcccat ggtggcggtc tctcgggcta 1080
gtttacggta gatccgtcac tgacctgccg atcaggcaaa cctattacat gggcacagag 1140
tgcgatgcgt cagacgtatc ggtgagtacg aacagcttat tgatggcttc ttataatgat 1200
ataggaactg ttccttactg gaaagggctt gaagccggtg agcccttcga aggctatacc 1260
ccagcaggaa tgacactcgc ggctggggag cgtattgtcc cgaaccacga atttcagatc 1320
tccgacgcca tggtacaagc agcgcagcgc caactagagg ctgtgcataa tcagaagcaa 1380
ctgcctcagc cctactcagc cgtttatcaa gaatggggtc acgagccata cggcggaggg 1440
tggcatgaat ggaaagcagg tttcgagtta gataaggtca tgcagaaaat gcgacacccg 1500
gtagaaggcg aggacatata tattgtggga gaagcgtact cgtatgatca agggtgggtt 1560
gagggtgctt tggtcgtagc cgaaagtatg cttgaggaat tttactgtct cgacgcacct 1620
agctggctaa aggtgtctga tgcgtcccat gctttcctgc cctcattatg caactgggtt 1680
ccacagttgc ttacggagcc gtatcctgtc cccgaaaatg actcggccac tgatgcaaac 1740
gcggctatcc gggccaccct caatgaggta acaacgtttg catacgaagg cataaaccac 1800
gcgcaacag 1809
<210>19
<211>1809
<212>DNA
<213>人工
<400>19
atggattcta ataaccaatc ccctcaggtt ttagaccaag ctattatcgg tggcggagtc 60
tcaggggtat attcggcctg gcgtttgcag caacagagtg gtggcgaaca aagcagtgca 120
ctttttgagt actctaatcg cattggaggg cgactctatt cccggaaaat ccccggtcta 180
gataacgtgg ttgcggaact gggcggaatg agatggatag ctgaggacca tccaatggtc 240
aattcattag tacacgaatt gaagcttgcc attaaagagt tcccgatggg gtcgagtctc 300
cctctagatc ccaaggtgaa agacagccca aaggcaggta ctgaaaacaa tctgttttac 360
ttaaggggcc agtatttccg ttaccgcgat cgtgcggagt gtccggaccg aatcccttat 420
aacttgggat ggtctgaacg ggggtacggt cccgaggata tgcaagttaa agtcatgaat 480
cttatatgcc caggcttcgc tgacatgtcc ctcgccgaac agatggcagt agaggtgaac 540
ggaaagccga tgtggcaata tgggttttgg aatctactgg aatcagtttt atcgaacgag 600
gcgtaccagt tcatgaaaga tgctggtggc tatgaagcca atgtcgcaaa cgcgaatgct 660
gtaacccaat tgcctgccac agagtacaag gacaacacgc agtttcttgg actcaaacaa 720
gggttccagg cactacccct gactttatgt gattgctttg aaaagttggg tggcgcggtg 780
catatggaca tgagacttgc tgagattagg atcgatccag ccagtgacac ccgttataca 840
ctcatattcc aaccgacgag cactgatgac tctggaaaaa ccacagatac ggacgatgca 900
tgtgttgaag tccaggcgaa gaaagtaatt ctagctatgc ctcgccgatc cctggagtta 960
atcaagtcag actactttga agatgagtgg ttgcaatcga atatacccag tgtgaagatt 1020
cagaaagcct tcaagatgtt tatggcatat gaaagcccat ggtggcggtc tctcgggcta 1080
gtttacggta gatccgtcac tgacctgccg atcaggcaaa cctattacat gggcacagag 1140
tgcgatgcgt cagacgtatc ggtgagtacg aacagcttat tgatggcttc ttataatgat 1200
ataggaactg ttccttactg gaaagggctt gaagccggtg agcccttcga aggctatacc 1260
ccagcaggaa tgacactcgc ggctggggag cgtattgtcc cgaaccacga atttcagatc 1320
tccgacgcca tggtacaagc agcgcagcgc caactagagg ctgtgcataa tcagaagcaa 1380
ctgcctcagc cctactcagc cgtttatcaa gaatggggtc acgagccata cggcggaggg 1440
tggcatgaat ggaaagcagg tttcgagtta gataaggtca tgcagaaaat gcgacacccg 1500
gtagaaggcg aggacatata tattgtggga gaagcgtact cgtatgatca agggtgggtt 1560
gagggtgctt tggtcgtagc cgaaagtatg cttgaggaat tttactgtct cgacgcacct 1620
agctggctaa aggtgtctga tgcgtcccat gctttcctgc cctcattatg caactgggtt 1680
ccacagttgc ttacggagcc gtatcctgtc cccgaaaatg actcggccac tgatgcaaac 1740
gcggctatcc gggccaccct caatgaggta acaacgtttg catacgaagg cataaaccac 1800
gcgcaacag 1809
<210>20
<211>1809
<212>DNA
<213>人工
<400>20
atggattcta ataaccaatc ccctcaggtt ttagaccaag ctattatcgg tggcggagtc 60
tcaggggtat attcggcctg gcgtttgcag caacagagtg gtggcgaaca aagcagtgca 120
ctttttgagt actctaatcg cattggaggg cgactctatt cccggaaaat ccccggtcta 180
gataacgtgg ttgcggaact gggcggaatg agatggatag ctgaggacca tccaatggtc 240
aattcattag tacacgaatt gaagcttgcc attaaagagt tcccgatggg gtcgagtctc 300
cctctagatc ccaaggtgaa agacagccca aaggcaggta ctgaaaacaa tctgttttac 360
ttaaggggcc agtatttccg ttaccgcgat tatgcggagt gtccggaccg aatcccttat 420
aacttgggat ggtctgaacg ggggtacggt cccgaggata tgcaagttaa agtcatgaat 480
cttatatgcc caggcttcgc tgacatgtcc ctcgccgaac agatggcagt agaggtgaac 540
ggaaagccga tgtggcaata tgggttttgg aatctactgg aatcagtttt atcgaacgag 600
gcgtaccagt tcatgaaaga tgctggtggc tatgaagcca atgtcgcaaa cgcgaatgct 660
gtaacccaat tgcctgccac agagtacaag gacaacacgc agtttcttgg actcaaacaa 720
gggttccagg cactacccct gactttatgt gattgctttg aaaagttggg tggcgcggtg 780
catatggaca tgagacttgc tgagattagg atcgatccag ccagtgacac ccgttataca 840
ctcatattcc aaccgacgag cactgatgac tctggaaaaa ccacagatac ggacgatgca 900
tgtgttgaag tccaggcgaa gaaagtaatt ctagctatgc ctcgccgatc cctggagtta 960
atcaagtcag actactttga agatgagtgg ttgcaatcga atatacccag tgtgcttatt 1020
cagaaagcct tcaagatgtt tatggcatat gaaagcccat ggtggcggtc tctcgggcta 1080
gtttacggta gatccgtcac tgacctgccg atcaggcaaa cctattacat gggcacagag 1140
tgcgatgcgt cagacgtatc ggtgagtacg aacagcttat tgatggcttc ttataatgat 1200
ataggaactg ttccttactg gaaagggctt gaagccggtg agcccttcga aggctatacc 1260
ccagcaggaa tgacactcgc ggctggggag cgtattgtcc cgaaccacga atttcagatc 1320
tccgacgcca tggtacaagc agcgcagcgc caactagagg ctgtgcataa tcagaagcaa 1380
ctgcctcagc cctactcagc cgtttatcaa gaatggggtc acgagccata cggcggaggg 1440
tggcatgaat ggaaagcagg tttcgagtta gataaggtca tgcagaaaat gcgacacccg 1500
gtagaaggcg aggacatata tattgtggga gaagcgtact cgtatgatca agggtgggtt 1560
gagggtgctt tggtcgtagc cgaaagtatg cttgaggaat tttactgtct cgacgcacct 1620
agctggctaa aggtgtctga tgcgtcccat gctttcctgc cctcattatg caactgggtt 1680
ccacagttgc ttacggagcc gtatcctgtc cccgaaaatg actcggccac tgatgcaaac 1740
gcggctatcc gggccaccct caatgaggta acaacgtttg catacgaagg cataaaccac 1800
gcgcaacag 1809
<210>21
<211>1809
<212>DNA
<213>人工
<400>21
atggattcta ataaccaatc ccctcaggtt ttagaccaag ctattatcgg tggcggagtc 60
tcaggggtat attcggcctg gcgtttgcag caacagagtg gtgacgaaca aagcagtgca 120
ctttttgagt actctaatcg cattggaggg cgactctatt cccggaaaat ccccggtcta 180
gataacgtgg ttgcggaact gggcggaatg agatggatag ctgaggacca tccaatggtc 240
aattcattag tacacgaatt gaagcttgcc attaaagagt tcccgatggg gtcgagtctc 300
cctctagatc ccaaggtgaa agacagccca aaggcaggta ctgaaaacaa tctgttttac 360
ttaaggggcc agtatttccg ttaccgcgat tatgcggagt gtccggaccg aatcccttat 420
aacttgggat ggtctgaacg ggggtacggt cccgaggata tgcaagttaa agtcatgaat 480
cttatatgcc caggcttcgc tgacatgtcc ctcgccgaac agatggcagt agaggtgaac 540
ggaaagccga tgtggcaata tgggttttgg aatctactgg aatcagtttt atcgaacgag 600
gcgtaccagt tcatgaaaga tgctggtggc tatgaagcca atgtcgcaaa cgcgaatgct 660
gtaacccaat tgcctgccac agagtacaag gacaacacgc agtttcttgg actcaaacaa 720
gggttccagg cactacccct gactttatgt gattgctttg aaaagttggg tggcgcggtg 780
catatggaca tgagacttgc tgagattagg atcgatccag ccagtgacac ccgttataca 840
ctcatattcc aaccgacgag cactgatgac tctggaaaaa ccacagatac ggacgatgca 900
tgtgttgaag tccaggcgaa gaaagtaatt ctagctatgc ctcgccgatc cctggagtta 960
atcaagtcag actactttga agatgagtgg ttgcaatcga atatacccag tgtgcagatt 1020
cagaaagcct tcaagatgtt tatggcatat gaaagcccat ggtggcggtc tctcgggcta 1080
gtttacggta gatccgtcac tgacctgccg atcaggcaaa cctattacat gggcacagag 1140
tgcgatgcgt cagacgtatc ggtgagtacg aacagcttat tgatggcttc ttataatgat 1200
ataggaactg ttccttactg gaaagggctt gaagccggtg agcccttcga aggctatacc 1260
ccagcaggaa tgacactcgc ggctggggag cgtattgtcc cgaaccacga atttcagatc 1320
tccgacgcca tggtacaagc agcgcagcgc caactagagg ctgtgcataa tcagaagcaa 1380
ctgcctcagc cctactcagc cgtttatcaa gaatggggtc acgagccata cggcggaggg 1440
tggcatgaat ggaaagcagg tttcgagtta gataaggtca tgcagaaaat gcgacacccg 1500
gtagaaggcg aggacatata tattgtggga gaagcgtact cgtatgatca agggtgggtt 1560
gagggtgctt tggtcgtagc cgaaagtatg cttgaggaat tttactgtct cgacgcacct 1620
agctggctaa aggtgtctga tgcgtcccat gctttcctgc cctcattatg caactgggtt 1680
ccacagttgc ttacggagcc gtatcctgtc cccgaaaatg actcggccac tgatgcaaac 1740
gcggctatcc gggccaccct caatgaggta acaacgtttg catacgaagg cataaaccac 1800
gcgcaacag 1809
<210>22
<211>1809
<212>DNA
<213>人工
<400>22
atggattcta ataaccaatc ccctcaggtt ttagaccaag ctattatcgg tggcggagtc 60
tcaggggtat attcggcctg gcgtttgcag caacagagtg gtgacgaaca aagcagtgca 120
ctttttgagt actctaatcg cattggaggg cgactctatt cccggaaaat ccccggtcta 180
gataacgtgg ttgcggaact gggcggaatg agatggatag ctgaggacca tccaatggtc 240
aattcattag tacacgaatt gaagcttgcc attaaagagt tcccgatggg gtcgagtctc 300
cctctagatc ccaaggtgaa agacagccca aaggcaggta ctgaaaacaa tctgttttac 360
ttaaggggcc agtatttccg ttaccgcgat cgtgcggagt gtccggaccg aatcccttat 420
aacttgggat ggtctgaacg ggggtacggt cccgaggata tgcaagttaa agtcatgaat 480
cttatatgcc caggcttcgc tgacatgtcc ctcgccgaac agatggcagt agaggtgaac 540
ggaaagccga tgtggcaata tgggttttgg aatctactgg aatcagtttt atcgaacgag 600
gcgtaccagt tcatgaaaga tgctggtggc tatgaagcca atgtcgcaaa cgcgaatgct 660
gtaacccaat tgcctgccac agagtacaag gacaacacgc agtttcttgg actcaaacaa 720
gggttccagg cactacccct gactttatgt gattgctttg aaaagttggg tggcgcggtg 780
catatggaca tgagacttgc tgagattagg atcgatccag ccagtgacac ccgttataca 840
ctcatattcc aaccgacgag cactgatgac tctggaaaaa ccacagatac ggacgatgca 900
tgtgttgaag tccaggcgaa gaaagtaatt ctagctatgc ctcgccgatc cctggagtta 960
atcaagtcag actactttga agatgagtgg ttgcaatcga atatacccag tgtgcagatt 1020
cagaaagcct tcaagatgtt tatggcatat gaaagcccat ggtggcggtc tctcgggcta 1080
gtttacggta gatccgtcac tgacctgccg atcaggcaaa cctattacat gggcacagag 1140
tgcgatgcgt cagacgtatc ggtgagtacg aacagcttat tgatggcttc ttataatgat 1200
ataggaactg ttccttactg gaaagggctt gaagccggtg agcccttcga aggctatacc 1260
ccagcaggaa tgacactcgc ggctggggag cgtattgtcc cgaaccacga atttcagatc 1320
tccgacgcca tggtacaagc agcgcagcgc caactagagg ctgtgcataa tcagaagcaa 1380
ctgcctcagc cctactcagc cgtttatcaa gaatggggtc acgagccata cggcggaggg 1440
tggcatgaat ggaaagcagg tttcgagtta gataaggtca tgcagaaaat gcgacacccg 1500
gtagaaggcg aggacatata tattgtggga gaagcgtact cgtatgatca agggtgggtt 1560
gagggtgctt tggtcgtagc cgaaagtatg cttgaggaat tttactgtct cgacgcacct 1620
agctggctaa aggtgtctga tgcgtcccat gctttcctgc cctcattatg caactgggtt 1680
ccacagttgc ttacggagcc gtatcctgtc cccgaaaatg actcggccac tgatgcaaac 1740
gcggctatcc gggccaccct caatgaggta acaacgtttg catacgaagg cataaaccac 1800
gcgcaacag 1809
<210>23
<211>1809
<212>DNA
<213>人工
<400>23
atggattcta ataaccaatc ccctcaggtt ttagaccaag ctattatcgg tggcggagtc 60
tcaggggtat attcggcctg gcgtttgcag caacagagtg gtgacgaaca aagcagtgca 120
ctttttgagt actctaatcg cattggaggg cgactctatt cccggaaaat ccccggtcta 180
gataacgtgg ttgcggaact gggcggaatg agatggatag ctgaggacca tccaatggtc 240
aattcattag tacacgaatt gaagcttgcc attaaagagt tcccgatggg gtcgagtctc 300
cctctagatc ccaaggtgaa agacagccca aaggcaggta ctgaaaacaa tctgttttac 360
ttaaggggcc agtatttccg ttaccgcgat tatgcggagt gtccggaccg aatcccttat 420
aacttgggat ggtctgaacg ggggtacggt cccgaggata tgcaagttaa agtcatgaat 480
cttatatgcc caggcttcgc tgacatgtcc ctcgccgaac agatggcagt agaggtgaac 540
ggaaagccga tgtggcaata tgggttttgg aatctactgg aatcagtttt atcgaacgag 600
gcgtaccagt tcatgaaaga tgctggtggc tatgaagcca atgtcgcaaa cgcgaatgct 660
gtaacccaat tgcctgccac agagtacaag gacaacacgc agtttcttgg actcaaacaa 720
gggttccagg cactacccct gactttatgt gattgctttg aaaagttggg tggcgcggtg 780
catatggaca tgagacttgc tgagattagg atcgatccag ccagtgacac ccgttataca 840
ctcatattcc aaccgacgag cactgatgac tctggaaaaa ccacagatac ggacgatgca 900
tgtgttgaag tccaggcgaa gaaagtaatt ctagctatgc ctcgccgatc cctggagtta 960
atcaagtcag actactttga agatgagtgg ttgcaatcga atatacccag tgtgaagatt 1020
cagaaagcct tcaagatgtt tatggcatat gaaagcccat ggtggcggtc tctcgggcta 1080
gtttacggta gatccgtcac tgacctgccg atcaggcaaa cctattacat gggcacagag 1140
tgcgatgcgt cagacgtatc ggtgagtacg aacagcttat tgatggcttc ttataatgat 1200
ataggaactg ttccttactg gaaagggctt gaagccggtg agcccttcga aggctatacc 1260
ccagcaggaa tgacactcgc ggctggggag cgtattgtcc cgaaccacga atttcagatc 1320
tccgacgcca tggtacaagc agcgcagcgc caactagagg ctgtgcataa tcagaagcaa 1380
ctgcctcagc cctactcagc cgtttatcaa gaatggggtc acgagccata cggcggaggg 1440
tggcatgaat ggaaagcagg tttcgagtta gataaggtca tgcagaaaat gcgacacccg 1500
gtagaaggcg aggacatata tattgtggga gaagcgtact cgtatgatca agggtgggtt 1560
gagggtgctt tggtcgtagc cgaaagtatg cttgaggaat tttactgtct cgacgcacct 1620
agctggctaa aggtgtctga tgcgtcccat gctttcctgc cctcattatg caactgggtt 1680
ccacagttgc ttacggagcc gtatcctgtc cccgaaaatg actcggccac tgatgcaaac 1740
gcggctatcc gggccaccct caatgaggta acaacgtttg catacgaagg cataaaccac 1800
gcgcaacag 1809
<210>24
<211>1809
<212>DNA
<213>人工
<400>24
atggattcta ataaccaatc ccctcaggtt ttagaccaag ctattatcgg tggcggagtc 60
tcaggggtat attcggcctg gcgtttgcag caacagagtg gtggcgaaca aagcagagca 120
ctttttgagt actctaatcg cattggaggg cgactctatt cccggaaaat ccccggtcta 180
gataacgtgg ttgcggaact gggcggaatg agatggatag ctgaggacca tccaatggtc 240
aattcattag tacacgaatt gaagcttgcc attaaagagt tcccgatggg gtcgagtctc 300
cctctagatc ccaaggtgaa agacagccca aaggcaggta ctgaaaacaa tctgttttac 360
ttaaggggcc agtatttccg ttaccgcgat cgtgcggagt gtccggaccg aatcccttat 420
aacttgggat ggtctgaacg ggggtacggt cccgaggata tgcaagttaa agtcatgaat 480
cttatatgcc caggcttcgc tgacatgtcc ctcgccgaac agatggcagt agaggtgaac 540
ggaaagccga tgtggcaata tgggttttgg aatctactgg aatcagtttt atcgaacgag 600
gcgtaccagt tcatgaaaga tgctggtggc tatgaagcca atgtcgcaaa cgcgaatgct 660
gtaacccaat tgcctgccac agagtacaag gacaacacgc agtttcttgg actcaaacaa 720
gggttccagg cactacccct gactttatgt gattgctttg aaaagttggg tggcgcggtg 780
catatggaca tgagacttgc tgagattagg atcgatccag ccagtgacac ccgttataca 840
ctcatattcc aaccgacgag cactgatgac tctggaaaaa ccacagatac ggacgatgca 900
tgtgttgaag tccaggcgaa gaaagtaatt ctagctatgc ctcgccgatc cctggagtta 960
atcaagtcag actactttga agatgagtgg ttgcaatcga atatacccag tgtgcagatt 1020
cagaaagcct tcaagatgtt tatggcatat gaaagcccat ggtggcggtc tctcgggcta 1080
gtttacggta gatccgtcac tgacctgccg atcaggcaaa cctattacat gggcacagag 1140
tgcgatgcgt cagacgtatc ggtgagtacg aacagcttat tgatggcttc ttataatgat 1200
ataggaactg ttccttactg gaaagggctt gaagccggtg agcccttcga aggctatacc 1260
ccagcaggaa tgacactcgc ggctggggag cgtattgtcc cgaaccacga atttcagatc 1320
tccgacgcca tggtacaagc agcgcagcgc caactagagg ctgtgcataa tcagaagcaa 1380
ctgcctcagc cctactcagc cgtttatcaa gaatggggtc acgagccata cggcggaggg 1440
tggcatgaat ggaaagcagg tttcgagtta gataaggtca tgcagaaaat gcgacacccg 1500
gtagaaggcg aggacatata tattgtggga gaagcgtact cgtatgatca agggtgggtt 1560
gagggtgctt tggtcgtagc cgaaagtatg cttgaggaat tttactgtct cgacgcacct 1620
agctggctaa aggtgtctga tgcgtcccat gctttcctgc cctcattatg caactgggtt 1680
ccacagttgc ttacggagcc gtatcctgtc cccgaaaatg actcggccac tgatgcaaac 1740
gcggctatcc gggccaccct caatgaggta acaacgtttg catacgaagg cataaaccac 1800
gcgcaacag 1809
<210>25
<211>1809
<212>DNA
<213>人工
<400>25
atggattcta ataaccaatc ccctcaggtt ttagaccaag ctattatcgg tggcggagtc 60
tcaggggtat attcggcctg gcgtttgcag caacagagtg gtggcgaaca aagcagagca 120
ctttttgagt actctaatcg cattggaggg cgactctatt cccggaaaat ccccggtcta 180
gataacgtgg ttgcggaact gggcggaatg agatggatag ctgaggacca tccaatggtc 240
aattcattag tacacgaatt gaagcttgcc attaaagagt tcccgatggg gtcgagtctc 300
cctctagatc ccaaggtgaa agacagccca aaggcaggta ctgaaaacaa tctgttttac 360
ttaaggggcc agtatttccg ttaccgcgat tatgcggagt gtccggaccg aatcccttat 420
aacttgggat ggtctgaacg ggggtacggt cccgaggata tgcaagttaa agtcatgaat 480
cttatatgcc caggcttcgc tgacatgtcc ctcgccgaac agatggcagt agaggtgaac 540
ggaaagccga tgtggcaata tgggttttgg aatctactgg aatcagtttt atcgaacgag 600
gcgtaccagt tcatgaaaga tgctggtggc tatgaagcca atgtcgcaaa cgcgaatgct 660
gtaacccaat tgcctgccac agagtacaag gacaacacgc agtttcttgg actcaaacaa 720
gggttccagg cactacccct gactttatgt gattgctttg aaaagttggg tggcgcggtg 780
catatggaca tgagacttgc tgagattagg atcgatccag ccagtgacac ccgttataca 840
ctcatattcc aaccgacgag cactgatgac tctggaaaaa ccacagatac ggacgatgca 900
tgtgttgaag tccaggcgaa gaaagtaatt ctagctatgc ctcgccgatc cctggagtta 960
atcaagtcag actactttga agatgagtgg ttgcaatcga atatacccag tgtgaagatt 1020
cagaaagcct tcaagatgtt tatggcatat gaaagcccat ggtggcggtc tctcgggcta 1080
gtttacggta gatccgtcac tgacctgccg atcaggcaaa cctattacat gggcacagag 1140
tgcgatgcgt cagacgtatc ggtgagtacg aacagcttat tgatggcttc ttataatgat 1200
ataggaactg ttccttactg gaaagggctt gaagccggtg agcccttcga aggctatacc 1260
ccagcaggaa tgacactcgc ggctggggag cgtattgtcc cgaaccacga atttcagatc 1320
tccgacgcca tggtacaagc agcgcagcgc caactagagg ctgtgcataa tcagaagcaa 1380
ctgcctcagc cctactcagc cgtttatcaa gaatggggtc acgagccata cggcggaggg 1440
tggcatgaat ggaaagcagg tttcgagtta gataaggtca tgcagaaaat gcgacacccg 1500
gtagaaggcg aggacatata tattgtggga gaagcgtact cgtatgatca agggtgggtt 1560
gagggtgctt tggtcgtagc cgaaagtatg cttgaggaat tttactgtct cgacgcacct 1620
agctggctaa aggtgtctga tgcgtcccat gctttcctgc cctcattatg caactgggtt 1680
ccacagttgc ttacggagcc gtatcctgtc cccgaaaatg actcggccac tgatgcaaac 1740
gcggctatcc gggccaccct caatgaggta acaacgtttg catacgaagg cataaaccac 1800
gcgcaacag 1809
<210>26
<211>1809
<212>DNA
<213>人工
<400>26
atggattcta ataaccaatc ccctcaggtt ttagaccaag ctattatcgg tggcggagtc 60
tcaggggtat attcggcctg gcgtttgcag caacagagtg gtggcgaaca aagcagtgca 120
ctttttgagt actctaatcg cattggaggg cgactctatt cccggaaaat ccccggtcta 180
gataacgtgg ttgcggaact gggcggaatg agatggatag ctgaggacca tccaatggtc 240
aattcattag tacacgaatt gaagcttgcc attaaagagt tcccgatggg gtcgagtctc 300
cctctagatc ccaaggtgaa agacagccca aaggcaggta ctgaaaacaa tctgttttac 360
ttaaggggcc agtatttccg ttaccgcgat cgtgcggagt gtccggaccg aatcccttat 420
aacttgggat ggtctgaacg ggggtacggt cccgaggata tgcaagttaa agtcatgaat 480
cttatatgcc caggcttcgc tgacatgtcc ctcgccgaac agatggcagt agaggtgaac 540
ggaaagccga tgtggcaata tgggttttgg aatctactgg aatcagtttt atcgaacgag 600
gcgtaccagt tcatgaaaga tgctggtggc tatgaagcca atgtcgcaaa cgcgaatgct 660
gtaacccaat tgcctgccac agagtacaag gacaacacgc agtttcttgg actcaaacaa 720
gggttccagg cactacccct gactttatgt gattgctttg aaaagttggg tggcgcggtg 780
catatggaca tgagacttgc tgagattagg atcgatccag ccagtgacac ccgttataca 840
ctcatattcc aaccgacgag cactgatgac tctggaaaaa ccacagatac ggacgatgca 900
tgtgttgaag tccaggcgaa gaaagtaatt ctagctatgc ctcgccgatc cctggagtta 960
atcaagtcag actactttga agatgagtgg ttgcaatcga atatacccag tgtgaagatt 1020
cagaaagcct tcaagatgtt tatggcatat gaaagcccat ggtggcggtc tctcgggcta 1080
gtttacggta gatccgtcac tgacctgccg atcaggcaaa cctattacat gggcacagag 1140
tgcgatgcgt cagacgtatc ggtgagtacg aacagcttat tgatggcttc ttataatgat 1200
ataggaactg ttccttactg gaaagggctt gaagccggtg agcccttcga aggctatacc 1260
ccagcaggaa tgacactcgc ggctggggag cgtattgtcc cgaaccacga atttcagatc 1320
tccgacgcca tggtacaagc agcgcagcgc caactagagg ctgtgcataa tcagaagcaa 1380
ctgcctcagc cctactcagc cgtttatcaa gaatggggtc acgagccata cggcggaggg 1440
tggcatgaat ggaaagcagg tttcgagtta gataaggtca tgcagaaaat gcgacacccg 1500
gtagaaggcg aggacatata tattgtggga gaagcgtact cgtatgatca agggtgggtt 1560
gagggtgctt tggtcgtagc cgaaagtatg cttgaggaat tttactgtct cgacgcacct 1620
agctggctaa aggtgtctga tgcgtcccat gctttcctgc cctcattatg caactgggtt 1680
ccacagttgc ttacggagcc gtatcctgtc cccgaaaatg actcggccac tgatgcaaac 1740
gcggctatcc gggccaccct caatgaggta acaacgtttg catacgaagg cataaaccac 1800
gcgcaacag 1809
<210>27
<211>1809
<212>DNA
<213>人工
<400>27
atggattcta ataaccaatc ccctcaggtt ttagaccaag ctattatcgg tggcggagtc 60
tcaggggtat attcggcctg gcgtttgcag caacagagtg gtgacgaaca aagcagagca 120
ctttttgagt actctaatcg cattggaggg cgactctatt cccggaaaat ccccggtcta 180
gataacgtgg ttgcggaact gggcggaatg agatggatag ctgaggacca tccaatggtc 240
aattcattag tacacgaatt gaagcttgcc attaaagagt tcccgatggg gtcgagtctc 300
cctctagatc ccaaggtgaa agacagccca aaggcaggta ctgaaaacaa tctgttttac 360
ttaaggggcc agtatttccg ttaccgcgat cgtgcggagt gtccggaccg aatcccttat 420
aacttgggat ggtctgaacg ggggtacggt cccgaggata tgcaagttaa agtcatgaat 480
cttatatgcc caggcttcgc tgacatgtcc ctcgccgaac agatggcagt agaggtgaac 540
ggaaagccga tgtggcaata tgggttttgg aatctactgg aatcagtttt atcgaacgag 600
gcgtaccagt tcatgaaaga tgctggtggc tatgaagcca atgtcgcaaa cgcgaatgct 660
gtaacccaat tgcctgccac agagtacaag gacaacacgc agtttcttgg actcaaacaa 720
gggttccagg cactacccct gactttatgt gattgctttg aaaagttggg tggcgcggtg 780
catatggaca tgagacttgc tgagattagg atcgatccag ccagtgacac ccgttataca 840
ctcatattcc aaccgacgag cactgatgac tctggaaaaa ccacagatac ggacgatgca 900
tgtgttgaag tccaggcgaa gaaagtaatt ctagctatgc ctcgccgatc cctggagtta 960
atcaagtcag actactttga agatgagtgg ttgcaatcga atatacccag tgtgcagatt 1020
cagaaagcct tcaagatgtt tatggcatat gaaagcccat ggtggcggtc tctcgggcta 1080
gtttacggta gatccgtcac tgacctgccg atcaggcaaa cctattacat gggcacagag 1140
tgcgatgcgt cagacgtatc ggtgagtacg aacagcttat tgatggcttc ttataatgat 1200
ataggaactg ttccttactg gaaagggctt gaagccggtg agcccttcga aggctatacc 1260
ccagcaggaa tgacactcgc ggctggggag cgtattgtcc cgaaccacga atttcagatc 1320
tccgacgcca tggtacaagc agcgcagcgc caactagagg ctgtgcataa tcagaagcaa 1380
ctgcctcagc cctactcagc cgtttatcaa gaatggggtc acgagccata cggcggaggg 1440
tggcatgaat ggaaagcagg tttcgagtta gataaggtca tgcagaaaat gcgacacccg 1500
gtagaaggcg aggacatata tattgtggga gaagcgtact cgtatgatca agggtgggtt 1560
gagggtgctt tggtcgtagc cgaaagtatg cttgaggaat tttactgtct cgacgcacct 1620
agctggctaa aggtgtctga tgcgtcccat gctttcctgc cctcattatg caactgggtt 1680
ccacagttgc ttacggagcc gtatcctgtc cccgaaaatg actcggccac tgatgcaaac 1740
gcggctatcc gggccaccct caatgaggta acaacgtttg catacgaagg cataaaccac 1800
gcgcaacag 1809
<210>28
<211>1809
<212>DNA
<213>人工
<400>28
atggattcta ataaccaatc ccctcaggtt ttagaccaag ctattatcgg tggcggagtc 60
tcaggggtat attcggcctg gcgtttgcag caacagagtg gtgacgaaca aagcagagca 120
ctttttgagt actctaatcg cattggaggg cgactctatt cccggaaaat ccccggtcta 180
gataacgtgg ttgcggaact gggcggaatg agatggatag ctgaggacca tccaatggtc 240
aattcattag tacacgaatt gaagcttgcc attaaagagt tcccgatggg gtcgagtctc 300
cctctagatc ccaaggtgaa agacagccca aaggcaggta ctgaaaacaa tctgttttac 360
ttaaggggcc agtatttccg ttaccgcgat tatgcggagt gtccggaccg aatcccttat 420
aacttgggat ggtctgaacg ggggtacggt cccgaggata tgcaagttaa agtcatgaat 480
cttatatgcc caggcttcgc tgacatgtcc ctcgccgaac agatggcagt agaggtgaac 540
ggaaagccga tgtggcaata tgggttttgg aatctactgg aatcagtttt atcgaacgag 600
gcgtaccagt tcatgaaaga tgctggtggc tatgaagcca atgtcgcaaa cgcgaatgct 660
gtaacccaat tgcctgccac agagtacaag gacaacacgc agtttcttgg actcaaacaa 720
gggttccagg cactacccct gactttatgt gattgctttg aaaagttggg tggcgcggtg 780
catatggaca tgagacttgc tgagattagg atcgatccag ccagtgacac ccgttataca 840
ctcatattcc aaccgacgag cactgatgac tctggaaaaa ccacagatac ggacgatgca 900
tgtgttgaag tccaggcgaa gaaagtaatt ctagctatgc ctcgccgatc cctggagtta 960
atcaagtcag actactttga agatgagtgg ttgcaatcga atatacccag tgtgaagatt 1020
cagaaagcct tcaagatgtt tatggcatat gaaagcccat ggtggcggtc tctcgggcta 1080
gtttacggta gatccgtcac tgacctgccg atcaggcaaa cctattacat gggcacagag 1140
tgcgatgcgt cagacgtatc ggtgagtacg aacagcttat tgatggcttc ttataatgat 1200
ataggaactg ttccttactg gaaagggctt gaagccggtg agcccttcga aggctatacc 1260
ccagcaggaa tgacactcgc ggctggggag cgtattgtcc cgaaccacga atttcagatc 1320
tccgacgcca tggtacaagc agcgcagcgc caactagagg ctgtgcataa tcagaagcaa 1380
ctgcctcagc cctactcagc cgtttatcaa gaatggggtc acgagccata cggcggaggg 1440
tggcatgaat ggaaagcagg tttcgagtta gataaggtca tgcagaaaat gcgacacccg 1500
gtagaaggcg aggacatata tattgtggga gaagcgtact cgtatgatca agggtgggtt 1560
gagggtgctt tggtcgtagc cgaaagtatg cttgaggaat tttactgtct cgacgcacct 1620
agctggctaa aggtgtctga tgcgtcccat gctttcctgc cctcattatg caactgggtt 1680
ccacagttgc ttacggagcc gtatcctgtc cccgaaaatg actcggccac tgatgcaaac 1740
gcggctatcc gggccaccct caatgaggta acaacgtttg catacgaagg cataaaccac 1800
gcgcaacag 1809
<210>29
<211>1809
<212>DNA
<213>人工
<400>29
atggattcta ataaccaatc ccctcaggtt ttagaccaag ctattatcgg tggcggagtc 60
tcaggggtat attcggcctg gcgtttgcag caacagagtg gtgacgaaca aagcagtgca 120
ctttttgagt actctaatcg cattggaggg cgactctatt cccggaaaat ccccggtcta 180
gataacgtgg ttgcggaact gggcggaatg agatggatag ctgaggacca tccaatggtc 240
aattcattag tacacgaatt gaagcttgcc attaaagagt tcccgatggg gtcgagtctc 300
cctctagatc ccaaggtgaa agacagccca aaggcaggta ctgaaaacaa tctgttttac 360
ttaaggggcc agtatttccg ttaccgcgat cgtgcggagt gtccggaccg aatcccttat 420
aacttgggat ggtctgaacg ggggtacggt cccgaggata tgcaagttaa agtcatgaat 480
cttatatgcc caggcttcgc tgacatgtcc ctcgccgaac agatggcagt agaggtgaac 540
ggaaagccga tgtggcaata tgggttttgg aatctactgg aatcagtttt atcgaacgag 600
gcgtaccagt tcatgaaaga tgctggtggc tatgaagcca atgtcgcaaa cgcgaatgct 660
gtaacccaat tgcctgccac agagtacaag gacaacacgc agtttcttgg actcaaacaa 720
gggttccagg cactacccct gactttatgt gattgctttg aaaagttggg tggcgcggtg 780
catatggaca tgagacttgc tgagattagg atcgatccag ccagtgacac ccgttataca 840
ctcatattcc aaccgacgag cactgatgac tctggaaaaa ccacagatac ggacgatgca 900
tgtgttgaag tccaggcgaa gaaagtaatt ctagctatgc ctcgccgatc cctggagtta 960
atcaagtcag actactttga agatgagtgg ttgcaatcga atatacccag tgtgaagatt 1020
cagaaagcct tcaagatgtt tatggcatat gaaagcccat ggtggcggtc tctcgggcta 1080
gtttacggta gatccgtcac tgacctgccg atcaggcaaa cctattacat gggcacagag 1140
tgcgatgcgt cagacgtatc ggtgagtacg aacagcttat tgatggcttc ttataatgat 1200
ataggaactg ttccttactg gaaagggctt gaagccggtg agcccttcga aggctatacc 1260
ccagcaggaa tgacactcgc ggctggggag cgtattgtcc cgaaccacga atttcagatc 1320
tccgacgcca tggtacaagc agcgcagcgc caactagagg ctgtgcataa tcagaagcaa 1380
ctgcctcagc cctactcagc cgtttatcaa gaatggggtc acgagccata cggcggaggg 1440
tggcatgaat ggaaagcagg tttcgagtta gataaggtca tgcagaaaat gcgacacccg 1500
gtagaaggcg aggacatata tattgtggga gaagcgtact cgtatgatca agggtgggtt 1560
gagggtgctt tggtcgtagc cgaaagtatg cttgaggaat tttactgtct cgacgcacct 1620
agctggctaa aggtgtctga tgcgtcccat gctttcctgc cctcattatg caactgggtt 1680
ccacagttgc ttacggagcc gtatcctgtc cccgaaaatg actcggccac tgatgcaaac 1740
gcggctatcc gggccaccct caatgaggta acaacgtttg catacgaagg cataaaccac 1800
gcgcaacag 1809
<210>30
<211>1809
<212>DNA
<213>人工
<400>30
atggattcta ataaccaatc ccctcaggtt ttagaccaag ctattatcgg tggcggagtc 60
tcaggggtat attcggcctg gcgtttgcag caacagagtg gtggcgaaca aagcagagca 120
ctttttgagt actctaatcg cattggaggg cgactctatt cccggaaaat ccccggtcta 180
gataacgtgg ttgcggaact gggcggaatg agatggatag ctgaggacca tccaatggtc 240
aattcattag tacacgaatt gaagcttgcc attaaagagt tcccgatggg gtcgagtctc 300
cctctagatc ccaaggtgaa agacagccca aaggcaggta ctgaaaacaa tctgttttac 360
ttaaggggcc agtatttccg ttaccgcgat cgtgcggagt gtccggaccg aatcccttat 420
aacttgggat ggtctgaacg ggggtacggt cccgaggata tgcaagttaa agtcatgaat 480
cttatatgcc caggcttcgc tgacatgtcc ctcgccgaac agatggcagt agaggtgaac 540
ggaaagccga tgtggcaata tgggttttgg aatctactgg aatcagtttt atcgaacgag 600
gcgtaccagt tcatgaaaga tgctggtggc tatgaagcca atgtcgcaaa cgcgaatgct 660
gtaacccaat tgcctgccac agagtacaag gacaacacgc agtttcttgg actcaaacaa 720
gggttccagg cactacccct gactttatgt gattgctttg aaaagttggg tggcgcggtg 780
catatggaca tgagacttgc tgagattagg atcgatccag ccagtgacac ccgttataca 840
ctcatattcc aaccgacgag cactgatgac tctggaaaaa ccacagatac ggacgatgca 900
tgtgttgaag tccaggcgaa gaaagtaatt ctagctatgc ctcgccgatc cctggagtta 960
atcaagtcag actactttga agatgagtgg ttgcaatcga atatacccag tgtgaagatt 1020
cagaaagcct tcaagatgtt tatggcatat gaaagcccat ggtggcggtc tctcgggcta 1080
gtttacggta gatccgtcac tgacctgccg atcaggcaaa cctattacat gggcacagag 1140
tgcgatgcgt cagacgtatc ggtgagtacg aacagcttat tgatggcttc ttataatgat 1200
ataggaactg ttccttactg gaaagggctt gaagccggtg agcccttcga aggctatacc 1260
ccagcaggaa tgacactcgc ggctggggag cgtattgtcc cgaaccacga atttcagatc 1320
tccgacgcca tggtacaagc agcgcagcgc caactagagg ctgtgcataa tcagaagcaa 1380
ctgcctcagc cctactcagc cgtttatcaa gaatggggtc acgagccata cggcggaggg 1440
tggcatgaat ggaaagcagg tttcgagtta gataaggtca tgcagaaaat gcgacacccg 1500
gtagaaggcg aggacatata tattgtggga gaagcgtact cgtatgatca agggtgggtt 1560
gagggtgctt tggtcgtagc cgaaagtatg cttgaggaat tttactgtct cgacgcacct 1620
agctggctaa aggtgtctga tgcgtcccat gctttcctgc cctcattatg caactgggtt 1680
ccacagttgc ttacggagcc gtatcctgtc cccgaaaatg actcggccac tgatgcaaac 1740
gcggctatcc gggccaccct caatgaggta acaacgtttg catacgaagg cataaaccac 1800
gcgcaacag 1809
<210>31
<211>1809
<212>DNA
<213>人工
<400>31
atggattcta ataaccaatc ccctcaggtt ttagaccaag ctattatcgg tggcggagtc 60
tcaggggtat attcggcctg gcgtttgcag caacagagtg gtgacgaaca aagcagagca 120
ctttttgagt actctaatcg cattggaggg cgactctatt cccggaaaat ccccggtcta 180
gataacgtgg ttgcggaact gggcggaatg agatggatag ctgaggacca tccaatggtc 240
aattcattag tacacgaatt gaagcttgcc attaaagagt tcccgatggg gtcgagtctc 300
cctctagatc ccaaggtgaa agacagccca aaggcaggta ctgaaaacaa tctgttttac 360
ttaaggggcc agtatttccg ttaccgcgat cgtgcggagt gtccggaccg aatcccttat 420
aacttgggat ggtctgaacg ggggtacggt cccgaggata tgcaagttaa agtcatgaat 480
cttatatgcc caggcttcgc tgacatgtcc ctcgccgaac agatggcagt agaggtgaac 540
ggaaagccga tgtggcaata tgggttttgg aatctactgg aatcagtttt atcgaacgag 600
gcgtaccagt tcatgaaaga tgctggtggc tatgaagcca atgtcgcaaa cgcgaatgct 660
gtaacccaat tgcctgccac agagtacaag gacaacacgc agtttcttgg actcaaacaa 720
gggttccagg cactacccct gactttatgt gattgctttg aaaagttggg tggcgcggtg 780
catatggaca tgagacttgc tgagattagg atcgatccag ccagtgacac ccgttataca 840
ctcatattcc aaccgacgag cactgatgac tctggaaaaa ccacagatac ggacgatgca 900
tgtgttgaag tccaggcgaa gaaagtaatt ctagctatgc ctcgccgatc cctggagtta 960
atcaagtcag actactttga agatgagtgg ttgcaatcga atatacccag tgtgaagatt 1020
cagaaagcct tcaagatgtt tatggcatat gaaagcccat ggtggcggtc tctcgggcta 1080
gtttacggta gatccgtcac tgacctgccg atcaggcaaa cctattacat gggcacagag 1140
tgcgatgcgt cagacgtatc ggtgagtacg aacagcttat tgatggcttc ttataatgat 1200
ataggaactg ttccttactg gaaagggctt gaagccggtg agcccttcga aggctatacc 1260
ccagcaggaa tgacactcgc ggctggggag cgtattgtcc cgaaccacga atttcagatc 1320
tccgacgcca tggtacaagc agcgcagcgc caactagagg ctgtgcataa tcagaagcaa 1380
ctgcctcagc cctactcagc cgtttatcaa gaatggggtc acgagccata cggcggaggg 1440
tggcatgaat ggaaagcagg tttcgagtta gataaggtca tgcagaaaat gcgacacccg 1500
gtagaaggcg aggacatata tattgtggga gaagcgtact cgtatgatca agggtgggtt 1560
gagggtgctt tggtcgtagc cgaaagtatg cttgaggaat tttactgtct cgacgcacct 1620
agctggctaa aggtgtctga tgcgtcccat gctttcctgc cctcattatg caactgggtt 1680
ccacagttgc ttacggagcc gtatcctgtc cccgaaaatg actcggccac tgatgcaaac 1740
gcggctatcc gggccaccct caatgaggta acaacgtttg catacgaagg cataaaccac 1800
gcgcaacag 1809
<210>32
<211>39
<212>DNA
<213>人工
<400>32
tcagctgtcg acatggattc taataaccaa tcccctcag 39
<210>33
<211>41
<212>DNA
<213>人工
<400>33
actgctgcta gcctgttgcg cgtggtttat gccttcgtat g 41
<210>34
<211>39
<212>DNA
<213>人工
<400>34
tcagctggat ccatggattc taataaccaa tcccctcag 39
<210>35
<211>41
<212>DNA
<213>人工
<400>35
actgctagat ctctgttgcg cgtggtttat gccttcgtat g 41
<210>36
<211>39
<212>DNA
<213>人工
<400>36
tcagctggta ccatggattc taataaccaa tcccctcag 39
<210>37
<211>38
<212>DNA
<213>人工
<400>37
actgctaagc ttctgttgcg cgtggtttat gccttcgtat g 41
<210>38
<211>39
<212>DNA
<213>人工
<400>38
tcagctatcg atatggattc taataaccaa tcccctcag 39
<210>39
<211>41
<212>DNA
<213>人工
<400>39
actgctcata tgctgttgcg cgtggtttat gccttcgtat g 41
Claims (12)
1.一种突变体,其特征在于,其氨基酸序列为SEQ ID NO.1上发生下列位点的突变:
Q332K;
Q332K、G34D;
Q332K、S38R;
Q332K、Y128R;
Q332K、G34D、S38R;
Q332K、G34D、Y128R;
Q332K、S38R、Y128R;
Q332K、G34D、S38R、Y128R。
2.根据权利要求1所述突变体,其特征在于,所述突变体对应的氨基酸序列分别为SEQID NO.5、SEQ ID NO.8、SEQ ID NO.10、SEQ ID NO.11、SEQ ID NO.13、SEQ ID NO.14、SEQID NO.15、SEQ ID NO.16。
3.一种编码权利要求2所述突变体的基因序列,其特征在于,所述基因序列被配置为SEQ ID NO.20、SEQ ID NO.23、SEQ ID NO.25、SEQ ID NO.26、SEQ ID NO.28、SEQ IDNO.29、SEQ ID NO.30、SEQ ID NO.31中的任一种,其中:
编码突变位点为Q332K的突变体的核酸序列为SEQ ID NO.20;
编码突变位点为G34D和Q332K的突变体的核酸序列为SEQ ID NO.23;
编码突变位点为S38R和Q332K的突变体的核酸序列为SEQ ID NO.25;
编码突变位点为Y128R和Q332K的突变体的核酸序列为SEQ ID NO.26;
编码突变位点为G34D、S38R和Q332K的突变体的精氨酸氧化酶突变体的核酸序列为SEQID NO.28;
编码突变位点为S38R、Y128R和Q332K的突变体的核酸序列为SEQ ID NO.29;
编码突变位点为G34D、Y128R和Q332K的突变体的核酸序列为SEQ ID NO.30;
编码突变位点为G34D、S38R、Y128R和Q332K的突变体的核酸序列为SEQ ID NO.31。
4.根据权利要求3所述基因序列,其特征在于,所述突变位点G34D的扩增引物对的核酸序列为SEQ ID NO.32和SEQ ID NO.33。
5.根据权利要求3所述基因序列,其特征在于,所述突变位点S38R的扩增引物对的核酸序列为SEQ ID NO.34和SEQ ID NO.35。
6.根据权利要求3所述基因序列,其特征在于,所述突变位点Y128R的扩增引物对的核酸序列为SEQ ID NO.36和SEQ ID NO.37。
7.根据权利要求3所述基因序列,其特征在于,所述突变位点Q332K的扩增引物对的核酸序列为SEQ ID NO.38和SEQ ID NO.39。
8.一种可溶性蛋白,其特征在于,其被配置为包括权利要求1~2任一所述突变体。
9.一种固定化酶,其特征在于,其被配置为包括权利要求1~2任一所述突变体。
10.一种重组工程细胞,其特征在于,其被配置为包括权利要求1~2任一所述突变体的编码序列。
11.一种重组载体,其特征在于,其被配置为包括权利要求1~2任一所述突变体的编码序列。
12.权利要求1~2任一所述突变体、权利要求8所述的可溶性蛋白、权利要求9所述的固定化酶、权利要求10所述的重组工程细胞或权利要求11所述的重组载体在催化氧化L-精氨酸中的应用。
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