CN112813093B - 一种带有激活标签的诱导型Ac/Ds转座子载体pRI-5及其应用 - Google Patents

一种带有激活标签的诱导型Ac/Ds转座子载体pRI-5及其应用 Download PDF

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CN112813093B
CN112813093B CN202011644089.7A CN202011644089A CN112813093B CN 112813093 B CN112813093 B CN 112813093B CN 202011644089 A CN202011644089 A CN 202011644089A CN 112813093 B CN112813093 B CN 112813093B
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张世忠
马晓君
齐琦
郑成超
吴长艾
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Shandong Agricultural University
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Abstract

本发明公开了一种带有激活标签的诱导型Ac/Ds转座子载体pRI‑5及其应用。所述诱导型Ac/Ds转座子载体pRI‑5的核苷酸序列如SEQ ID No.23所示，包括Ac元件、Ds元件。所述诱导型Ac/Ds转座子载体pRI‑5可以通过地塞米松的诱导转化构建植物突变体库，进而筛选得到一种具有短根表型的小肽，该小肽的氨基酸序列如SEQ ID No.26所示，编码该小肽的基因A的核苷酸序列如SEQ ID No.25所示，在植株中超表达基因A或在植株外施合成的小肽均可使植物的根系变短。本发明通过少量的转基因植株即可诱导产生大量的Ds插入株系，成本低，节省人工转基因的工作量，适用性广。

Description

一种带有激活标签的诱导型Ac/Ds转座子载体pRI-5及其应用
技术领域
本发明属于植物转基因技术领域，具体涉及一种带有激活标签的诱导型Ac/Ds转座子载体pRI-5及其应用。
背景技术
转座子，又叫可移动因子，是染色体上一种依靠自身编码的酶通过复制或切离方式随机移动的DNA片段，当在植物染色体上某个基因内部或附近区域插入时，可能会改变基因的表达量而导致突变。在Ac/Ds转座子系统中，Ac元件具有合成转座酶的能力，但没有转座能力；Ds元件两端含有能被转座酶识别的反向重复序列和完整的转座序列，所以只有当Ac元件和Ds元件同时存在时才能发生转座。因此现在很多学者利用Ac/Ds转座子系统构建突变体库，也就是利用Ac/Ds转座子系统产生少量的转化株系就可以获得大量Ds插入突变体，节省人工转基因的工作量。然而，目前很多Ac/Ds转座子系统转化后的突变体库中的突变体多为隐形突变，这增加了基因筛选的难度。
与植物生长有关的植物肽激素是由5-60个氨基酸残基组成的，既能在组织液扩散中短距离发挥作用，也能通过导管和筛管长距离运输。肽激素具有序列短、浓度低、功能强的特点。在植物的生长发育中具有重要的作用，参与根、茎、叶、花、种子的发育，对生物胁迫和非生物胁迫的响应。目前，还有更多的植物肽激素等待着被发现，而通过突变体库的筛选也是很好的手段之一。
发明内容
本发明公开了一种带有激活标签的诱导型Ac/Ds转座子载体pRI-5及其应用。本发明通过诱导型Ac/Ds转座子载体pRI-5转化构建得到植株突变体库，并筛选到一种具有短根表型的小肽。
为实现上述发明目的，本发明采用以下技术方案予以实现：
本发明提供了一种带有激活标签的诱导型Ac/Ds转座子载体pRI-5，所述带有激活标签的诱导型Ac/Ds转座子载体pRI-5的核苷酸序列如SEQ ID No.23所示。
进一步的：所述带有激活标签的诱导型Ac/Ds转座子载体pRI-5包括Ac元件、Ds元件。
进一步的：所述Ac元件由35S启动子启动来合成Ac转座酶，所述Ac转座酶后带有GR。
进一步的：所述Ds元件内包括35S：：mgfp5-nos荧光片段和4×35S激活标签；所述Ds元件的两端分别含有35S片段和mCherry片段。
本发明还提供了所述的带有激活标签的诱导型Ac/Ds转座子载体pRI-5在用于构建植物突变体库中的应用。
进一步的：所述植物突变体库通过所述带有激活标签的诱导型Ac/Ds转座子载体pRI-5转化获得，具体步骤包括：
(1)将所述带有激活标签的诱导型Ac/Ds转座子载体pRI-5转入发根农杆菌后并活化，得到含目的基因的活化发根农杆菌菌液；
(2)将植物组培苗于继代培养基中，25℃，16h光照/8h黑暗的条件下培养30天后，切掉愈伤团，切口处在含目的基因的活化发根农杆菌菌液中蘸取一次，然后在pH 5.2的1/2MS培养基中22℃培养3天，转移到含500mg/L特美汀抗生素的pH5.9的1/2MS培养基中，每十天转移一次，直至组培苗长出转基因毛状根系；
(3)将荧光鉴定为绿色的转基因毛状根系带组培苗转移至含有5-10μm/L地塞米松的培养基中，25℃，16h光照/8h黑暗的条件下培养，直到荧光鉴定根系出现红色时，得到大量不同的Ds插入植物株系，即获得植物突变体库。
进一步的：所述植物包括苹果。
本发明还提供了一种调控植物根系变短的小肽，所述小肽是通过权利要求1所述的带有激活标签的诱导型Ac/Ds转座子载体pRI-5转化构建得到的植物突变体库中经筛选分离获得的。
进一步的：所述小肽的氨基酸序列如SEQ ID No.26所示。
进一步的：编码所述小肽的基因A的核苷酸序列如SEQ ID No.25所示。
进一步的：在植株外施合成的所述小肽或者在植株中超表达编码所述小肽的基因A都能够使植株的根系变短。
本发明与现有技术相比，具有以下优点和有益效果：
本发明构建得到带有激活标签的诱导型Ac/Ds转座子载体pRI-5，能够利用地塞米松诱导Ac转座酶表达，使其进入细胞核发挥作用，有利于得到稳定的Ds插入株系，建立良好的植物突变体库；且利用红色荧光指示Ds转座情况，Ds一经转座即可检测到，步骤简单快捷，结果观察明显。诱导型Ac/Ds转座子载体pRI-5还通过加入激活标签，促使Ds插入位点附近基因表达，有利于出现新表型，进而筛选到显性突变体。本发明还从诱导型Ac/Ds转座子载体pRI-5转化得到的苹果突变体库中筛选得到一种具有短根表型的突变体，其编码得到的小肽合成后外施或超表达在植株中，都可以使苹果的根系变短。本发明通过少量的转基因植株即可诱导产生大量的Ds插入株系，成本低，该方法适用性广，是一种理想的植物基因组学研究工具。
附图说明
图1：转基因株系绿色荧光检测。
图2：Ds插入株系红色荧光检测。
图3：超表达小肽根系表型对比图。
图4：小肽外施根系表型对比图。
具体实施方式
以下结合具体实施例对本发明的技术方案做进一步详细的说明。
实施例1
带有激活标签的诱导型Ac/Ds转座子载体的制备，包括以下步骤：
(1)使用pSQ5转基因小麦(Triticum aestivum)的cDNA扩增得到Ac元件(GenBankNo：X05424)；在pTA7002载体中扩增得到糖皮质激素受体(GR)；在pRI101-AN中扩增得到nos和35S片段；在pSQ5载体中扩增得到3Ds元件、5Ds元件和4×35S增强子；pBI121-mgfp5和pmRi-mCherry载体中分别扩增得到35S：：mgfp5-nos和mCherry荧光片段。
其中，pSQ5载体通过参考文献(Qu SH et al.A versatile transposon-basedactivation tag vector system for functional genomics in cereals and othermonocot plants.2008.Plant Physiol，190-199.)获得。
pBI121-mgfp5载体为改造载体，即将原始pBI121载体中的GUS元件替换为mgfp5荧光载体。
扩增所使用的酶为来自南京诺唯赞生物科技股份有限公司的Phanta Max Super-Fidelity DNA Polymerase。扩增条件为95℃ 3min；95℃ 15s，50-60℃ 15s，72℃ 30s/kb，重复35个循环；72℃ 5min。扩增体系(50μL体系)：2×Phanta Max Buffer 25μL，dNTP Mix(10mM each)1μL，上游引物(10μM)2μL，下游引物(10μM)2μL，Phanta Max Super-FidelityDNA Polymerase1μL，模板DNA 2μL，ddH2O补充到50μL。
使用的扩增引物如下所示：
AC-F：cgatcggggaaattcgagctcATGACGCCTCCGGTTGGA(SEQ ID No.1)；
AC-R：cttgctgaattggatcTGGAGAGGAGCCACTTGCTACA(SEQ ID No.2)；
GR-F：aagtggctcctctccaGATCCAATTCAGCAAGCCACTG(SEQ ID No.3)；
GR-R：cccgtcgaccccgggggtaccTCATTTTTGATGAAACAGAAGCTTTT(SEQ ID No.4)；
nos-F：cccgtcgaccccgggggtaccGATCGTTCAAACATTTGGCAATAA(SEQ ID No.5)；
nos-R：cctttgttgaaaagtctcaGATCTAGTAACATAGATGACACCGCG(SEQ ID No.6)；
35s-F：tctatgttactagatcTGAGACTTTTCAACAAAGGGTAATATC(SEQ ID No.7)；
35S-R：agagttgttgattcagaattcTGTTCTCTCCAAATGAAATGAACTTC(SEQ ID No.8)；
3D-F：gggggtaccggatccgaattcTAGGGATGAAAACGGTCGGTAA(SEQ ID No.9)；
3D-R：ccctttgttgaaaagtctcaACTAGTAATATATGTTTTCATGTGTGATTTTACCG(SEQ IDNo.10)；
4×35S-F：tgttgaaaagtctcaactagtTAGTGGATCCCCAACATGGTG(SEQ ID No.11)；
4×35S-R：atgaaaacatatattactagtTCGGATCTAGATATCACATCAATCCACT(SEQ IDNo.12)；
mgfp-F：catgaaaacatatattTGAGACTTTTCAACAAAGGGTAATATC(SEQ ID No.13)；
mgfp-R：cgatcggggaaattcgagctcGATCTAGTAACATAGATGACACCGCG(SEQ ID No.14)；
5D-F：tcgggaattgctaccgagctcAAGCTTCTTTAGGCTAACCACTTCA(SEQ ID No.15)；
5D-R：tgctcaccatCAGGGATGAAAGTAGGATGGGA(SEQ ID No.16)；
mCherry-F：ttcatccctgATGGTGAGCAAGGGCGAGG(SEQ ID No.17)；
mCherry-R：cgatcggggaaattcgagctcTTACTTGTACAGCTCGTCCATGCC(SEQ IDNo.18)。
(2)将上述扩增得到的元件或片段进行两两同源重组，重组为Ac+GR、nos+35S、3Ds+35S：：mgfp5-nos和5Ds+mCherry，然后将这些重组片段连接到pRI101-AN载体中，对应的酶切位点分别是Sal I和Kpn I、Kpn I和EcoR I、EcoR I和Sac I以及单酶切位点Sac I，由此获得中间载体pRI-1、pRI-2、pRI-3和pRI-4。
同源重组使用的是苏州新赛美生物科技有限公司的快速同源重组克隆试剂盒，反应条件为50℃反应30min；反应体系(10μL体系)：2×EasyFusion Assembly Master mix 5μL，线性化克隆载体10-50ng，插入片段20-50ng，ddH2O补充到10μL。
载体酶切使用的是宝日医生物技术(北京)有限公司的限制性内切酶，反应条件为37℃反应3h；反应体系(10μL体系)：Buffer 2μL，酶1 1μL，酶2 1μL，载体10μL，ddH2O补充到10μL。
(3)使用pRI-3和pRI-4的扩增引物对在中间载体pRI-3和pRI-4上扩增获得3Ds+35S：：mgfp5-nos和5Ds+mCherry片段，再通过同源重组连接到中间载体pRI-2上，酶切位点是EcoR I和Sac I，由此获得中间载体pRI-234。
使用的扩增引物如下所示：
pRI-3-F：catttggagagaacagaattcTAGGGATGAAAACGGTCGGTAA(SEQ ID No.19)；
pRI-3-R：gcctaaagaagcttATCTAGTAACATAGATGACACCGCG(SEQ ID No.20)；
pRI-4-F：tacaaacacgacgaactctaaAAGCTTCTTTAGGCTAACCACTTCA(SEQ ID No.21)；
pRI-4-R：cgatcggggaaattcgagctcTTACTTGTACAGCTCGTCCATGCC(SEQ ID No.22)。
(4)使用Sal I和Kpn I限制性内切酶将重组片段Ac+GR从pRI-1上切下，再使用T4连接酶将片段Ac-GR连接到中间载体pRI-234上，由此得到诱导型Ac/Ds转座子载体pRI-1234；
(5)在pRI-1234的3Ds后含有酶切位点Spe I，酶切后再通过同源重组将4×35S增强子插入到pRI-1234载体中，从而获得带有激活标签的双荧光Ac/Ds转座子载体pRI-5，其核苷酸序列如SEQ ID No.23所示。
由于转座子载体pRI-5中含有Ac元件，并由35S启动子启动Ac元件表达来合成Ac转座酶，而Ac转座酶后带有糖皮质激素受体(GR)，所以当地塞米松(糖皮质激素配体)存在时Ac转座酶才能进入细胞核发挥作用。
转座子载体pRI-5中含有不可自主转座的Ds元件，Ds内含有35S：：mgfp5-nos荧光片段，所以当转座子载体pRI-5转入植株后能检测到绿色荧光(图1)，且Ds内含有4×35S激活标签，当Ds插入后能激活Ds插入位点附近的基因表达；另外，Ds两端分别含有35S片段和mCherry片段，因此当Ds转座后能检测到红色荧光(图2)。
荧光检测使用的是LUYOR-3415激发光源，观察绿色荧光使用的是蓝光照射，佩戴LUV-30A黄色眼镜；观察红色荧光使用的是绿光照射，佩戴LUV-50A红色眼镜。
实施例2
利用带有激活标签的诱导型Ac/Ds转座子载体pRI-5转化苹果砧木平邑甜茶，包括以下步骤：
(1)将转座子载体pRI-5转入发根农杆菌后并活化，得到含目的基因的活化发根农杆菌菌液；
(2)将生长状态良好的平邑甜茶的组培苗放入继代培养基(MS培养基+蔗糖30g/L+6-BA 0.5mg/L+IBA 0.05mg/L+琼脂7g/L)中，于25℃，16h光照/8h黑暗的条件下培养30天，得到继代组培苗；
(3)将继代组培苗的叶片切去前后两端，将叶片在活化好的发根农杆菌菌液中蘸取一次，叶片远轴面向下在pH 5.2的1/2MS培养基(1/2MS培养液+蔗糖30g/L+琼脂6g/L)中22℃共培养3天；
(4)将侵染叶片转移到含500mg/L特美汀抗生素的pH为5.9的1/2MS培养基中培养，每十天转移一次，直到长出转基因毛状根系；
(5)将长出转基因毛状根系的侵染组培苗放回生根培养基(1/2MS培养液+蔗糖25g/L+IBA 0.5mg/L+琼脂6g/L)中，于28℃，16h光照/8h黑暗条件下共培养1-2天后，再插到含500mg/L特美汀抗生素的1/2MS培养基中培养，直到长出大量的转基因毛状根，并通过绿色荧光鉴定这些转基因根系；
(6)将带有大量转基因根系的侵染组培苗转移到含有5-10μm/L地塞米松的1/2MS培养基中，于25℃，16h光照/8h黑暗的条件下培养，当检测到转基因根系有红色荧光时说明Ds元件发生转座，此时获得大量不同的Ds插入苹果株系，即得到苹果突变体库。
实施例3
1、在所述苹果突变体库中找到一个具有根系变短的表型的突变体。通过hiTAIL-PCR实验分离DS侧翼序列，DS插入位置如SEQ ID No.24所示，其中在该序列中1-1483bp和6363-6879bp为启动子，1484-6362bp为插入序列，6880-6942和7232-7440bp为外显子，6943-7231bp为内含子，并发现该突变是由于Ds插入基因A的启动子区域引起的，插入基因A启动子区域后，导致基因A表达增加，进而导致突变体表现为短根表型。基因A的核苷酸序列如SEQ ID No.25所示，该基因A编码得到了含有76个氨基酸的小肽，小肽的氨基酸序列如SEQ ID No.26所示。
使用hiTAIL-PCR实验分离侧翼序列的具体流程如下：
(1)预扩增PCR反应体系：
Figure BDA0002878543780000071
(2)第一轮PCR反应体系：
Figure BDA0002878543780000072
(3)第二轮PCR反应体系：
Figure BDA0002878543780000073
(4)hiTAIL-PCR反应程序如下：
Figure BDA0002878543780000074
Figure BDA0002878543780000081
(5)hiTAIL-PCR使用引物如下：
LAD1-1：ACGATGGACTCCAGCGGCCGCVNVNNNGGAA(SEQ ID No.27)；
LAD1-2：ACGATGGACTCCAGCGGCCGCBNBNNNGGTT(SEQ ID No.28)；
LAD1-3：ACGATGGACTCCAGCGGCCGCHNVNNNCCAC(SEQ ID No.29)；
LAD1-4：ACGATGGACTCCAGCGGCCGCVVNVNNNCCAA(SEQ ID No.30)；
AC1：ACGATGGACTCCAGAG(SEQ ID No.31)；
3DS-0：GTCTCAAATATATGTTTTCATGTGTGATTTTACCGAACAAAAATACCGGT(SEQ IDNo.32)；
3DS-1：
ACGATGGACTCCAGTCCGGCCCGTCCGATTTCGACTTTAACCCGACCGGATCGTATCGG(SEQ IDNo.33)；
3DS-2：CCCGTTCGTTTTCGTTACCGGTATATCCCGTTTTCGTTTCCGTCC(SEQ ID No.34)；
其中，引物LAD1-1、LAD1-2、LAD1-3、LAD1-4为兼并引物与3Ds-0一起用于预扩增PCR反应，引物3Ds-1用于第一轮PCR反应，引物3Ds-2第二轮PCR反应。
2、将基因A在植株中超表达会导致根系变短，具体流程如下：
(1)使用平邑甜茶(Malus hupehensis)的cDNA扩增得到基因A序列，使用Ac/Ds转座子载体pRI-5扩增得到nos-35S序列，通过同源重组将这两个片段连接到载体pBI121-mgfp5上，酶切位点为Xba I和BamH I，由此得到35S：：A-35S：：mgfp5双表达核载体。
其中，使用的扩增引物如下所示：
A-F：gagaacacgggggactctagaATGGCGACATTGGTGAGGAA(SEQ ID No.35)；
A-R：ttgaacgatcTCATGGAGGGCAAGGCTTG(SEQ ID No.36)；
Nos-35S-F：ccctccatgaGATCGTTCAAACATTTGGCAATAA(SEQ ID No.37)；
Nos-35S-R：agtcttcatacccggggatccTGTTCTCTCCAAATGAAATGAACTTC(SEQ IDNo.38)。
(2)将35S：：A-35S：：mgfp5双表达核载体转入发根农杆菌后并活化，得到含目的基因的活化发根农杆菌菌液；
(3)将生长状态良好的平邑甜茶的组培苗放入继代培养基中，于25℃，16h光照/8h黑暗的条件下培养30天，得到继代组培苗；
(4)切下继代组培苗的3-4cm茎段，在活化好的发根农杆菌菌液中浸泡8min，无菌水冲洗一次，在滤纸上吸干多余水分，将此茎段放入含有500mg/L特美汀抗生素的pH 5.9的1/2MS培养基中，25℃培养，直到长出转基因根系，观察其表型。结果如图3所示，与野生型相比，超表达基因A的植株表现出根系变短的表型。
3、所述小肽合成外施同样会导致根系变短，具体流程如下：
(1)在生工生物工程(上海)股份有限公司根据氨基酸序列合成本发明的小肽；
(2)选择大小相似的平邑甜茶种子，使用次氯酸钠消毒杀菌14min，无菌水清洗6次，将种子上的水分在滤纸上吸干后铺到1/2MS培养基中，在4℃黑暗环境下层积35-40d；
(3)取萌发一致，约1cm的平邑甜茶苗转移到含有1μM/L小肽的1/2MS培养基中，25℃，16h光照/8h黑暗的条件下培养至根生长到板底，统计根长表型。结果如图4所示，与野生型(未添加小肽)的植株相比，外施小肽的植株明显的表现出根系变短的表型。
因此，通过在植株内超表达基因A或者在植株外施合成的小肽都可以使得植株的根系变短，说明通过带有激活标签的双荧光Ac/Ds转座子载体pRI-5可以建立苹果突变体库，进而筛选到能够直接应用于农业生产中的突变体或小肽，且筛选到的小肽有利于农业上的研究和发展。
以上实施例仅用以说明本发明的技术方案，而非对其进行限制；尽管参照前述实施例对本发明进行了详细的说明，对于本领域的普通技术人员来说，依然可以对前述实施例所记载的技术方案进行修改，或者对其中部分技术特征进行等同替换；而这些修改或替换，并不使相应技术方案的本质脱离本发明所要求保护的技术方案的精神和范围。
序列表
<110> 山东农业大学
<120> 一种带有激活标签的诱导型Ac/Ds转座子载体pRI-5及其应用
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tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 60
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aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 180
cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 240
gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 300
ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 360
cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 420
aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 480
tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 540
ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 600
tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 660
ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 720
agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 780
atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 840
cctatctcag cgatctgtct atttcgttca tccatagttg cctgactcga tcctacaagg 900
tagaatccgc ctgagtcgca agggtgactt cgcctatatt ggacgacggc gcgcagaggg 960
cgacctcttt ttgggttacg attgtaggat tatcactaaa acaatacatg aacatattca 1020
aatggcaatc tctctaaggc attggaaata aatacaaata acagttgggt ggagtttttc 1080
gacctgaggg cgttaacctt ctgttaacct aaaagctctt gcccaaacag cagaatcggc 1140
gctaattgcc agcggcggaa cttttccagt ttcgcgaaaa atatcgccac tggcaaggaa 1200
tgggtttgag atggcgaagt ctgtcctaaa agcagcgcct gtagttgtag ggttgacggc 1260
cttgatggag cgtcatgccg atgccctctc gagccaactt caagcacatc atcttaaggt 1320
tttcccgccg cattccgaga agggtattcg aacattcggg ccatcggagg cgtccaagct 1380
gctcggcgtt ggcgagtcat atttacggca gaccgcgtct gagatgccag agttgaatgt 1440
tagcatgagc ccaggtggca ggcgaatgtt ctcaattgaa gatatccatg tgattcggaa 1500
gtatatggat caggtcggcc gcgggaaccg gcgctacctg ccacatcgtc gaggcggcga 1560
gcagcttcag gttatctctg tgatgaattt caaaggtggg tcgggtaaga ccaccaccgc 1620
cgcgcatctg gcgcagtacc tcgctatgcg cggatatcga gtcttggcca ttgatctcga 1680
tcctcaagcg agcctttctg cactctttgg gagccaaccg gagacggacg ttggcccgaa 1740
cgaaacgctc tacggcgcta taaggtatga tgatgagcag gtggcaatcg aacgagtcgt 1800
ccgagggact tacattcccg acctccacct gattcctggt aaccttgagc tgatggagtt 1860
tgaacacgat acgccacgcg cgctgatgaa ccgcaaagag ggcgacacgc tcttttatgg 1920
tcgcatcagc caagtaattg aagatatcgc ggataactat gacgtcgtgg tcatcgactg 1980
ccctccccag cttgggtatc tcacgctatc cgcattgact gcggcgacgt ccattcttgt 2040
cacggtccat ccgcagatgc tggatgtgat gtcgatgaac cagtttctgg caatgacatc 2100
gaaccttttg cgtgaaatcg agaatgctgg cgccaagttc aagtttaatt ggatgcgcta 2160
tctgataacc cgtttcgaac cgagcgacgg accacagaac caaatggtag gttatctgcg 2220
gtcgattttt ggcgaaaatg tcctcaattt tccgatgctt aaaaccaccg cggtttcgga 2280
cgctggcctg acaaaccaga ctctattcga agtggagcgt ggcctgttca cgcgctcgac 2340
ctatgatcga gccttggagg cgatgaacgc cgtcaacgac gagatcgaaa cactgatcaa 2400
aaaagcatgg ggtaggccca catgagccgg aagcacatcc ttggcgtctc aactgacgcc 2460
cctgagacgt cgcccgccga caataggacg gcaaagaacc gctccatgcc gctcctcggc 2520
gtaacaagga aggagcgcga tccggcaacg aagctcacag cgaacattgg taacgcactg 2580
cgagagcaaa acgatcgtct tagccgtgcc gaagagatcg agcggcgtct cgctgaaggt 2640
caggcagtga tagagttgga tgcctcgtca atagaaccgt ctttcgtgca ggatcgtatg 2700
cgaggggaca ttgacgggct ccttacttcg atccgggaac aaggacagca agtcccaatc 2760
cttgtgcgac cgcatccgag ccagccgggc cgatatcagg ttgccttcgg ccaccgccgg 2820
ctacgcgccg tttcagaact cggacttccg gtcagggcgg tcgttcgcga actgacggac 2880
gagcaagtgg tcgtagcaca gggtcaggaa aacaatgtgc gcgaagatct taccttcatc 2940
gaaaaggcgc gcttcgcaca tcgcctgaac aggcagtttt ctcgagagat tgtcatcgcc 3000
gcgatgtcga tcgacaagag caatttgtcc aagatgcttc tgctcgttga cgccctcccc 3060
tctgaactga ccgatgctat tggtgccgct cctggtgttg gacggccgag ttggcaacaa 3120
cttgccgagc tgattgagaa agtttcttca ccggccgacg tggctaaata tgctatgtcg 3180
gaggaagttc aagcgctgcc atcggcagaa cgattcaagg cggtgatcgc tagtctgaag 3240
cccagtcggg ttgcgcgtgg acttcccgag gtcatggcca ccccagacgg caccagaatt 3300
gcacaggtga cgcagagcaa ggccaaactg gaaatcacga ttgacaggaa ggcgacgccc 3360
gattttgcga ccttcgtgct cgatcatgtg ccagcgctgt atcaagcgta ccacgctgag 3420
aaccaacgga aacggggaga gtaaaccgca aaagaaaaga gccccctcaa cgtcgccgtc 3480
gcggaagccc ttctgtctct ctagcgcgaa cagaatcgca tttcctcgaa tcctcgtcaa 3540
gagtttttag cgccgttttg gtgagctgat ttcctttgcc tgctgaaagg tgaaagatga 3600
tgcagacagg aagtgtaacg acgccattcg ggcggcggcc aatgacgctt gcgcttgtgc 3660
ggcgccagac ggcgctggcc gatatcaaac aaggcaagac agcggacaag tggaaggtct 3720
ttagggacgc gtccgcggcc atggaactac ttggaatcca gtccaacagt cttgccgtcc 3780
ttgatgcgct attgagcttt cacccggaaa cggagttgcg tcaggaggca cagctgatcg 3840
tcttcccgtc gaatgctcag cttgcccttc gggcgcatgg gatggctggc gcgactttgc 3900
gtaggcacat cgccatgctc gtggagtcag gcttgatcgt ccggaaggat agcgccaacg 3960
gaaagcgtta cgctcgtaag gatggcgctg gtcagatcga gcgcgcgttt ggcttcgatt 4020
tgtctccgct tctcgcgcgg tccgaagagc tagcgatgat ggcacagcag gtgatggccg 4080
atcgagcagc attcaggatg gccaaagaaa gtctgacgat ttgccgacgg gacgttcgga 4140
agctaattac ggcagctatg gaagagggag cggagggcga ctggcaagct gtcgaggaag 4200
tctatgtgga acttgtgggt agaatcccac gcgccccgac gcttgctgat gtagagtcaa 4260
ttctcgaaga gatgtggatg ctccaggaag agataatcaa ccggttggaa attagagaca 4320
attcagaaaa taatagcacc aatgctgccc agagcgagca gcacatacag aactcaaaac 4380
ccgaatccgt taatgaactt gaacctcgct ctgaaaagga gcagggcgct aagccgagtg 4440
aaatagaccg ggcaaggagc gagccgataa aagcgttccc cctcgggatg atcctgaaag 4500
cgtgcccgac cattggcaat tatgggccga gcggtgcggt tgctagctgg cgtgacctca 4560
tgtcggctgc ggtggtggtt cggtctatgc tgggggtcag cccgtcggct taccaagacg 4620
cgtgtgaggc aatgggaccg gagaatgcgg cagcagcgat ggcgtgcatt ttggagcgag 4680
cgaacttcat caattcgcct gggggctatc tccgagatct gacacggcgg agcgaactcg 4740
ggaagttttc acttggcccg atgataatgg cgctcttgaa ggctagcggg caggggacgt 4800
tgcggtttgg ctagaattag cgagtatgga gcaggatggt ctgtggtcag ctgaccacag 4860
acctaatagg ttgaaaacat gagcgttttt tggatgatcg acagaccatc cgattcccgg 4920
agtaccaagc gtgctctgat gggagcgata acattactca acaagcacga aggccccatg 4980
ccgatcgttg atcgtgaagg agagcctgct ctacatgcgg cggtattttg ccggccgagg 5040
catgtagtcg cggagcactg cctatttact gccctaggca caaacgttga ctcttggatc 5100
gagctggcag acaaagcaat aacccacaca gaggacgatt aatggctgac gaagagatcc 5160
agaatccgcc ggacggtact gctgctgccg aagttgagcc ggctgctcct agaggtagaa 5220
gagcaaagaa agcaccagcc gaaacagccc gcacgggatc gttcaaatcc gtgaagccga 5280
aaacccgcgg cctcagcaac cgagaaaaac tggagaagat cggtcaaatc gaagctcagg 5340
tcgctggcgg cgcaaccttg aaggacgccg ttaagatcgt gggtatttcc gttcagacct 5400
attatcaatg gaagagagct gcggttcaac ctgtctcaca gaatccggcc gtgtctgttt 5460
cagttgacga tgaactcggc gagttcatcc aactcgagga ggaaaatcgg cggctcagaa 5520
agctagcttg tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata 5580
acacattgcg gacgttttta atgtactggg gtggtttttc ttttcaccag tgagacgggc 5640
aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg 5700
gtttgcccca gcaggcgaaa atcctgtttg atggtggttc cgaaatcggc aaaatccctt 5760
ataaatcaaa agaatagccc gagatagggt tgagtgttgt tccagtttgg aacaagagtc 5820
cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg 5880
gcccaaattc ccgatctagt aacatagatg acaccgcgcg cgataattta tcctagtttg 5940
cgcgctatat tttgttttct atcgcgtatt aaatgtataa ttgcgggact ctaatcataa 6000
aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc ttaacgtaat 6060
tcaacagaaa ttatatgata atcatcgcaa gaccggcaac aggattcaat cttaagaaac 6120
tttattgcca aatgtttgaa cgatcggggc ccgatcatat tgtcgctcag gatcgtggcg 6180
ttgtgcttgt cggccgttgc tgtcgtaatg atatcggcac cttcgaccgc ctgttccgca 6240
gagatcccgt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg 6300
gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa 6360
tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc 6420
agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac 6480
cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg 6540
tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc 6600
cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga 6660
cgagatcatc gccgtcgggc atacgcgcct tgagcctggc gaacagttcg gctggcgcga 6720
gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac 6780
gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg 6840
tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag 6900
atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag 6960
tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg 7020
ctgcctcgtc ctgtagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg 7080
ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg 7140
cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat 7200
cttgttcaat catgcgaaac gatccagatc cggtgcagat tatttggatt gagagtgaat 7260
atgagactct aattggatac cgaggggaat ttatggaacg tcagtggagc atttttgaca 7320
agaaatattt gctagctgat agtgacctta ggcgactttt gaacgcgcaa taatggtttc 7380
tgacgtatgt gcttagctca ttaaactcca gaaacccgcg gctgagtggc tccttcaacg 7440
ttgcggttct gtcagttcca aacgtaaaac ggcttgtccc gcgtcatcgg cgggggtcat 7500
aacgtgactc ccttaattct ccgctcatga tcctggcacg acaggtttcc cgactggaaa 7560
gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct 7620
ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac 7680
acaggaaaca gctatgacca tgattacgaa ttaattcccg atctagtaac atagatgaca 7740
ccgcgcgcga taatttatcc tagtttgcgc gctatatttt gttttctatc gcgtattaaa 7800
tgtataattg cgggactcta atcataaaaa cccatctcat aaataacgtc atgcattaca 7860
tgttaattat tacatgctta acgtaattca acagaaatta tatgataatc atcgcaagac 7920
cggcaacagg attcaatctt aagaaacttt attgccaaat gtttgaacga tcggggaaat 7980
tcgagctctt acttgtacag ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt 8040
tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg 8100
acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt cttgacctca 8160
gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc cttcagggcg 8220
ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt cttctgcatt 8280
acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat gaactcgccg 8340
tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg 8400
cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat gtcggcgggg 8460
tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc ccaggcgaag 8520
ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg 8580
ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc 8640
ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt gctcaccatc 8700
agggatgaaa gtaggatggg aaaatcccgt accgaccgtt atcgtataac cgattttgtt 8760
agttttatcc cgatcgattt cgaacccgag gtaaaaaacg aaaacggaac ggaaacggga 8820
tatacaaaac ggtaaacgga aacggaaacg gtagagctag tttcccgacc gtttcaccgg 8880
gatcccgttt ttaatcggga tgatcccgtt tcgttaccgt attttctaat tcgggatgac 8940
tgcaatatgg ccagctccaa ctcccatcca taaccactga ggcccagccc atgtaagaaa 9000
tacctagcga acgctgctct gcctctctcc caggcggcca ggcaccacac gagtaacagc 9060
atcacacatt cacacgccgc cacgcgccca cgccggagtc cggacgccgc cagccgcacg 9120
ccgacgccgg cgacgcgtct cgctctcgcc tgctctctcc gactctccct gtctcccagc 9180
cggccggccg ctgggctgca ccaggcacca cacgcggtga cggccgtgac gcggcacgcc 9240
ggacgcagac gccgccatcc acggtccgcc ctccactcca ctgctcgcga ctcgcccatc 9300
cgcgccgcgg tccgcccatc cgcccagacc tccactccac tgctcgccca tccgcggtcc 9360
gcccatccgc catccgccat ctgcggtcag cggtcctcca gacctccact gctcggcgct 9420
cgcccatccg gccatccgcg gtctccctgt ctccacggct gctcacaggc tcacagcact 9480
tagcagtaca gcacgtcagc accattgcac caagctgttg tgtcatttgt gtgctgtcca 9540
ggggctctgc aacacctgct gattgctgtc cagccgtcca ggtgctcaca agtcacagca 9600
gtacagcacc aagctgattg ctgaacacct gctgtccagg gctctgctct ccacttcggc 9660
tagccggcta cgactccatt cctcagatga cgcctccggt tggaaataat cctccctcag 9720
gctcagccat aagattggcc aagttgatgt ctaccacaag agcgccttct actcgcaaaa 9780
caaattccgt attctctgca tatgctcaag gtatatatta gaaaaacagt agcaatagca 9840
ttagcattac taattggttg tagattggga agcatcatat tgactgtaga ataatacgaa 9900
aaatctgttt ataacagggt tgaaaagaaa agctgaagcc tcttctagtc ggattcagaa 9960
tgtacgtgca cgtgcgcgtg ggcatggatg tggccgcaca tcaccatcat catcaacagc 10020
tgaggccgag aggcatttta ttcagagtgt aagcagtagt aatgcaaatg gtacagctac 10080
agatccgagt caagatgata tggctattgt tcatgaacca caaccacaac cacaaccaca 10140
accagaacca caaccacagc cacaacctga acccgaagaa gaagcaccac agaagagggc 10200
aaagaagtgc acatcggatg tatggcagca tttcaccaag aaggaaattg aagtggaggt 10260
cgatggaaag aaatacgttc aggtatgggg acattgcaac tttcctaatt gcaaggctaa 10320
gtatagggct gagggtcatc atggaacaag cggatttcga aatcacttga gaacatcaca 10380
tagtttagtt aaaggtcagt tgtgtctaaa aagtgaaaag gatcatggca aagacataaa 10440
tctcattgag ccttataagt acgatgaagt ggttagccta aagaagcttg agctcggtag 10500
caattcccga ggctgtagcc gacgatggtg cgccaggaga gttgttgatt cagaattcca 10560
tcctacgatc tagtaacata gatgacaccg cgcgcgataa tttatcctag tttgcgcgct 10620
atattttgtt ttctatcgcg tattaaatgt ataattgcgg gactctaatc ataaaaaccc 10680
atctcataaa taacgtcatg cattacatgt taattattac atgcttaacg taattcaaca 10740
gaaattatat gataatcatc gcaagaccgg caacaggatt caatcttaag aaactttatt 10800
gccaaatgtt tgaacgatcg agctcttaga gttcgtcgtg tttgtatagt tcatccatgc 10860
catgtgtaat cccagcagct gttacaaact caagaaggac catgtggtct ctcttttcgt 10920
tgggatcttt cgaaagggcg gattgtgtgg acaggtaatg gttgtccggt aaaaggacag 10980
ggccatcgcc aattggagta ttttgttgat aatgatcagc gagttgcacg ccgccgtctt 11040
cgatgttgtg gcgggtcttg aagttggctt tgatgccgtt cttttgcttg tcggccatga 11100
tgtatacgtt gtgggagttg tagttgtatt ccaacttgtg accgaggatg tttccgtcct 11160
ccttgaaatc gattccctta agctcgatcc tgttgacgag ggtgtctccc tcaaacttga 11220
cttcagcacg tgtcttgtag ttcccgtcgt ccttgaagaa gatggtcctc tcctgcacgt 11280
atccctcagg catggcgctc ttgaagaagt cgtgccgctt catatgatct gggtatcttg 11340
aaaagcattg aacaccataa gagaaagtag tgacaagtgt tggccatgga acaggtagtt 11400
ttccagtagt gcaaataaat ttaagggtaa gttttccgta tgttgcatca ccttcaccct 11460
ctccactgac agaaaatttg tacccattaa catcaccatc taattcaaca agaattggga 11520
caactccagt gaaaagttct tctcctttac tgaattcggc cgaggataat gataggagaa 11580
gtgaaaagat gagaaagaga aaaagattag tcttcatacc cggggatcct ctagagtccc 11640
ccgtgttctc tccaaatgaa atgaacttcc ttatatagag gaagggtctt gcgaaggata 11700
gtgggattgt gcgtcatccc ttacgtcagt ggagatatca catcaatcca cttgctttga 11760
agacgtggtt ggaacgtctt ctttttccac gatgctcctc gtgggtgggg gtccatcttt 11820
gggaccactg tcggcagagg catcttcaac gatggccttt cctttatcgc aatgatggca 11880
tttgtaggag ccaccttcct tttccactat cttcacaata aagtgacaga tagctgggca 11940
atggaatccg aggaggtttc cggatattac cctttgttga aaagtctcaa ctagtggatc 12000
cccaacatgg tggagcacga cactctcgtc tactccaaga atatcaaaga tacagtctca 12060
gaagaccaga gggctattga gacttttcaa caaagggtaa tatcgggaaa cctcctcgga 12120
ttccattgcc cagctatctg tcacttcatc gaaaggacag tagaaaagga agatggcttc 12180
tacaaatgcc atcattgcga taaaggaaag gctatcgttc aagatgcctc taccgacagt 12240
ggtcccaaag atggaccccc acccacgagg aacatcgtgg aaaaagaaga cgttccaacc 12300
acgtcttcaa agcaagtgga ttgatgtgat atctagatcc ccaacatggt ggagcacgac 12360
actctcgtct actccaagaa tatcaaagat acagtctcag aagaccagag ggctattgag 12420
acttttcaac aaagggtaat atcgggaaac ctcctcggat tccattgccc agctatctgt 12480
cacttcatcg aaaggacagt agaaaaggaa gatggcttct acaaatgcca tcattgcgat 12540
aaaggaaagg ctatcgttca agatgcctct accgacagtg gtcccaaaga tggaccccca 12600
cccacgagga acatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat 12660
tgatgtgata tctagatccc caacatggtg gagcacgaca ctctcgtcta ctccaagaat 12720
atcaaagata cagtctcaga agaccagagg gctattgaga cttttcaaca aagggtaata 12780
tcgggaaacc tcctcggatt ccattgccca gctatctgtc acttcatcga aaggacagta 12840
gaaaaggaag atggcttcta caaatgccat cattgcgata aaggaaaggc tatcgttcaa 12900
gatgcctcta ccgacagtgg tcccaaagat ggacccccac ccacgaggaa catcgtggaa 12960
aaagaagacg ttccaaccac gtcttcaaag caagtggatt gatgtgatat ctagatcccc 13020
aacatggtgg agcacgacac tctcgtctac tccaagaata tcaaagatac agtctcagaa 13080
gaccagaggg ctattgagac ttttcaacaa agggtaatat cgggaaacct cctcggattc 13140
cattgcccag ctatctgtca cttcatcgaa aggacagtag aaaaggaaga tggcttctac 13200
aaatgccatc attgcgataa aggaaaggct atcgttcaag atgcctctac cgacagtggt 13260
cccaaagatg gacccccacc cacgaggaac atcgtggaaa aagaagacgt tccaaccacg 13320
tcttcaaagc aagtggattg atgtgatatc taactagtaa tatatgtttt catgtgtgat 13380
tttaccgaac aaaaataccg gttcccgtcc gatttcgact ttaacccgac cggatcgtat 13440
cggttttcga ttaccgtatt tatcccgttc gttttcgtta ccggtatatc ccgttttcgt 13500
ttccgtcccg caagttaaat atgaaaatga aaacggtaga ggtattttac cgaccgttac 13560
cgaccgtttt catccctaga attctgttct ctccaaatga aatgaacttc cttatataga 13620
ggaagggtct tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag tggagatatc 13680
acatcaatcc acttgctttg aagacgtggt tggaacgtct tctttttcca cgatgctcct 13740
cgtgggtggg ggtccatctt tgggaccact gtcggcagag gcatcttcaa cgatggcctt 13800
tcctttatcg caatgatggc atttgtagga gccaccttcc ttttccacta tcttcacaat 13860
aaagtgacag atagctgggc aatggaatcc gaggaggttt ccggatatta ccctttgttg 13920
aaaagtctca gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg 13980
cgctatattt tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa 14040
acccatctca taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc 14100
aacagaaatt atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt 14160
tattgccaaa tgtttgaacg atcggtacct catttttgat gaaacagaag ctttttgata 14220
tttccatttg aatattttgg tatctgatta gtgatgattt cagctaacat ctctgggaat 14280
tcaatactca tggtcttatc caaaaatgtc tggaagcagt aggtaaggag attctcaacc 14340
acctcatgca tggagtccag aagctttgtc agttggtaaa accgttgcca gttctgactg 14400
gagttccctt cccttttgac gatggctttt cctagctctt tgatataagt cattcgaatc 14460
tcatcaaata actcttggct cttcagacct tccttaggaa ctgaggagag aagcagtaag 14520
gttttcatac agagatactc ttcataggat acctgcaatc tttgtaattc agaggagaca 14580
aacagcatgt gtttacattg gtcatacatg cagggtagag acattctctg ctcattaata 14640
atcagatcag gagcaaagca gagcaggttt ccacttgatt gtctgtatga tctccaaccc 14700
agggcaaatg ccatgagaaa catccatgag tactgtagca gggtcatttg gtcatccagg 14760
tgtaagtttc tgaagcctgg tatcgccttt gcccatttca ctgctgcaat cacttgacgc 14820
ccacctaaca tgttgagtgt ggtcataatt ctccatgctg aatctggaac agagctatca 14880
tatcctgcat acaacacctc gggttcaatc acctccagca gtgacaccaa ggtaggggtg 14940
agctgtggta atgcagcagg aactattgtt ttgttaggat tttccgaagt gtcttgtgag 15000
actcctgcag tggcttgctg aattggatct ggagaggagc cacttgctac atcttcatta 15060
ttcttagaaa attctattgc gtcttcatcc tcatccatat gattctcatg atttgttgca 15120
gcagcaataa cagagtctag cacctcgaga tcaccaatca ttgttggaaa atatgtagca 15180
ccttttctag atgctgctac ccaatctttt gtgcatatca aagcttcaac aatctccgaa 15240
ccaagacgat tgcggtaagg atcaacaaca cgaccaccag cactgaacgc agactcagaa 15300
gcaacagttg acacttgtat tgctagcaca tcccttgcaa tttgggtgag aataggatat 15360
tctgcaaccc ttcccctcca ccatgataaa atatcaaact gaccactatg cttcaaaagg 15420
ggttcagaca tatatttatc caattcattt gactctactt gatcataatc cttcaactca 15480
tgcaaatagt tttgaaattc atcatcttca ttttccatca aggtatcatc catactatca 15540
ttagtagttg tctttgtctt tggagctgaa ggactacaac tagaatagaa ttgatacaat 15600
tttctaatga ccctaacaaa gtcatctaca tgaactttgt atgaatcacc atgaaatttt 15660
ttcatataga actcaatcaa tattttcttg tacctagggt caaggaagca tgctacagct 15720
agtgcaatat tagacacttt ccaatatttc tcaaactttt cactcattgc aacggccatt 15780
ctcctaatga caaatttttc atgaacacac cattggtcaa tcaaatcctt tatctcacag 15840
aaacctttgt aaaataaatt tgcagtggaa tattgagtac cagataggag ttcagtgaga 15900
tcaaaaaact tcttcaaaca cttaaaaaga gttaatgcca tcttccactc ctcggcttta 15960
ggacaaattg catcgtacct gcgaggatca cttgttttaa gccttattag tgcaggctta 16020
taatataagg catccctcaa catcaaatag gttgaattcc atctagttga gacatcatat 16080
gagatccctt tagatttatc caagtcacat tcactagcac acttcattag ttcttcccac 16140
tgcaaaggag aagattttac agcaagaaca atcgctttga ttttctcaat tgttcctgca 16200
attacagcca agccatcctt tgcaaccaag ttcagtatgt gacaagcaca cctcacatga 16260
aagaaagcac catcacaaac tagatttgaa tcagtgtcct gcaaatcctc aattatatcg 16320
tgcacagcta cttcatttgc actagcatta tccaaagaca aggcaaacaa ttttttctca 16380
atgttccact taaccatgat tgcagtgaag gtttgtgata acctttggcc agtgtggcgc 16440
ccttcaacat gaaaaaagcc aacaattctt ttttggagac accaatcatc atcaatccaa 16500
tggatggtga cacacatgta tgacttattt tgacaagatg tccacatatc catagttgta 16560
ctgaagcgag actgaacatc ttttagtttt ccatacaact tttctttttc ttccaaatac 16620
aaatccatga tatattttct agcagtgaca cgggacttta ttggaaagtg agggcgcaga 16680
gacttaacaa actcaacaaa gtactcatgt tctacaatat tgaaaggata ttcatgcatg 16740
attattgcca aatgaagctt ctttaggcta accacttcat cgtacttata aggctcaatg 16800
agatttatgt ctttgccatg atccttttca ctttttagac acaactgacc tttaactaaa 16860
ctatgtgatg ttctcaagtg atttcgaaat ccgcttgttc catgatgacc ctcagcccta 16920
tacttagcct tgcaattagg aaagttgcaa tgtccccata cctgaacgta tttctttcca 16980
tcgacctcca cttcaatttc cttcttggtg aaatgctgcc atacatccga tgtgcacttc 17040
tttgccctct tctgtggtgc ttcttcttcg ggttcaggtt gtggctgtgg ttgtggttct 17100
ggttgtggtt gtggttgtgg ttgtggttca tgaacaatag ccatatcatc ttgactcgga 17160
tctgtagctg taccatttgc attactactg cttacactct gaataaaatg cctctcggcc 17220
tcagctgttg atgatgatgg tgatgtgcgg ccacatccat gcccacgcgc acgtgcacgt 17280
acattctgaa tccgactaga agaggcttca gcttttcttt tcaacccttg agcatatgca 17340
gagaatacgg aatttgtttt gcgagtagaa ggcgctcttg tggtagacat caacttggcc 17400
aatcttatgg ctgagcctga gggaggatta tttccaaccg gaggcgtcat gtcgacgggc 17460
atatgtatca acagtgaaga acttgctttt gatcttttgt tagttttgtg tgattgtgat 17520
gtatctagag tcccccgtgt tctctccaaa tgaaatgaac ttccttatat agaggaaggg 17580
tcttgcgaag gatagtggga ttgtgcgtca tcccttacgt cagtggagat atcacatcaa 17640
tccacttgct ttgaagacgt ggttggaacg tcttcttttt ccacgatgct cctcgtgggt 17700
gggggtccat ctttgggacc actgtcggca gaggcatctt caacgatggc ctttccttta 17760
tcgcaatgat ggcatttgta ggagccacct tccttttcca ctatcttcac aataaagtga 17820
cagatagctg ggcaatggaa tccgaggagg tttccggata ttaccctttg ttgaaaagtc 17880
tcaattgccc tttggtcttc tgagactgta tctttgatat ttttggagta gacaagtgtg 17940
tcgtgctcca ccatgttgac gaagattttc ttcttgtcat tgagtcgtaa gagactctgt 18000
atgaactgtt cgccagtctt tacggcgagt tctgttaggt cctctatttg aatctttgac 18060
tccatggcct ttgattcagt gggaactacc tttttagaga ctccaatctc tattacttgc 18120
cttggtttgt gaagcaagcc ttgaatcgtc catactggaa tagtacttct gatcttgaga 18180
aatatatctt tctctgtgtt cttgatgcag ttagtcctga atcttttgac tgcatcttta 18240
accttcttgg gaaggtattt gatttcctgg agattattgc tcgggtagat cgtcttgatg 18300
agacctgctg cgtaagcctc tctaaccatc tgtgggttag cattctttct gaaattgaaa 18360
aggctaatct ggggacctgc aggcatgcaa gcttggcact ggccgtcgtt ttacaacgtc 18420
gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg 18480
ccaggatcag attgtcgttt cccgccttca gtttaaacta tcagtgtttg acaggatata 18540
ttggcgggta aacctaagag aaaagagcgt ttattagaat aatcggatat ttaaaagggc 18600
gtgaaaaggt ttatccgttc gtccatttgt atgtgactaa aacaattcat ccagtaaaat 18660
ataatatttt attttctccc aatcaggctt gatccccagt aagtcaaaaa atagctcgac 18720
atactgttct tccccgatat cctccctgat cgaccggacg cagaaggcaa tgtcatacca 18780
cttgtccgcc ctgccgcttc tcccaagatc aataaagcca cttactttgc catctttcac 18840
aaagatgttg ctgtctccca ggtcgccgtg ggaaaagaca agttcctctt cgggcttttc 18900
cgtctttaaa aaatcataca gctcgcgcgg atctttaaat ggagtgtctt cttcccagtt 18960
ttcgcaatcc acatcggcca gatcgttatt cagtaagtaa tccaattcgg ctaagcggct 19020
gtctaagcta ttcgtatagg gacaatccga tatgtcgatg gagtgaaaga gcctgatgca 19080
ctccgcatac agctcgataa tcttttcagg gctttgttca tcttcatact cttccgagca 19140
aaggacgcca tcggcctcac tcatgagcag attgctccag ccatcatgcc gttcaaagtg 19200
caggaccttt ggaacaggca gctttccttc cagccatagc atcatgtcct tttcccgttc 19260
cacatcatag gtggtccctt tataccggct gtccgtcatt tttaaatata ggttttcatt 19320
ttctcccacc agcttatata ccttagcagg agacattcct tccgtatctt ttacgcagcg 19380
gtatttttcg atcagttttt tcaattccgg tgatattctc attttagcca tttattattt 19440
ccttcctctt ttctacagta tttaaagata ccccaagaag ctaattataa caagacgaac 19500
tccaattcac tgttccttgc attctaaaac cttaaatacc agaaaacagc tttttcaaag 19560
ttgttttcaa agttggcgta taacatagta tcgacggagc cgattttgaa accacaatta 19620
tgggtgatgc tgccaactta ctgatttagt gtatgatggt gtttttgagg tgctccagtg 19680
gcttctgttt ctatcagctg tccctcctgt tcagctactg acggggtggt gcgtaacggc 19740
aaaagcaccg ccggacatca gcgctatctc tgctctcact gccgtaaaac atggcaac 19798
<210> 24
<211> 7467
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 24
gggtgaggtt gagcgtcgga gaggaagaga gagagaactg agagtgagag aaagtcgtga 60
gggaagagag atagagagag agagtgggtt acgggtttta gggaaataat tggggaggat 120
gggtcttttt aaatccataa tacaaccttt taacatacaa acttaaacct aagtccaaat 180
ttaagtttgt aatcctatat aatatcccag ttaacaatag tatagggaaa ttatgtaaaa 240
accaatgcca aagtgacgca ccttaatccc gtttaaggtt gtattggtca tttcacgtcc 300
tcggaattaa taattccggg acgggctgta acactatgac cctttgaccc tcggttggag 360
acggtttttt gtgacagggc taaaacgagc cctttggccc tctgaccctc ggttggagac 420
ggaggcaaat atgaccctat actgttcatt aaaatattaa tatcttgggg aatcttagag 480
ggccagaggg ctcaaacgag ccctctagcc agccctcggt tggagatggc tttaaatcct 540
taagtaaact acccaaccaa aaaaatctga aacagaactg tgaatcacat tattaaataa 600
caacaaaaat tgaggactta caaataaaca gaatgaaaat tgggatttat ttctaagcgg 660
tgttttgttt gggggattaa tgcatgctaa accctctggc gaggtttttt ttatttttta 720
tttttatttc tgtaggaaag ttgtctcatt gtaaattagg aaagaaaaca gaacattgaa 780
gttgttccat tgtaaattag ggaagaaaac agaaaattca agctgtccca ttataaatta 840
ggataaaaaa aaacttttta tattagcacc ccacaaaatg ttgaatgcac cccacattaa 900
aatttaatat aactaactta tttaccctac tatgcaatga caattttgat cttattaggt 960
ttaaaaaaaa aattctaaat acaccccaaa ttacttttaa cgtattattt atcttcttca 1020
actatcaaaa cccttttgac cctccattgt tgaacaaaac cctaactaat gaaattacaa 1080
acatattaat tgagaaaaat tattttttga cgaatgaaac acgaaatcat gttttggaag 1140
cttcacatga gataagactt catttttttt attgttttag tgatttgacg acatcgataa 1200
ttatttaatc ttgcccttgt tgcgttattc aagcttctat gtttgtattc atcttttagg 1260
ggtcgtaggg attatatgaa gaattaaaca cctaaaatta ccaccacata ttaaaaacag 1320
acgacatgag ttttaatggt ggaattgaaa ataacccaca tatgctttaa aggcagcatc 1380
ttttgtcaaa attctagttg acattttttg tccaaaataa aagctttata agatttgaga 1440
aatgtggtgt gtatagaaaa tatggggcgc atgtagaaaa tgtcagggat gaaagtagga 1500
tgggaaaatc ccgtaccgac cgttatcgta taaccgattt tgttagtttt atcccgatcg 1560
atttcgaacc cgaggtaaaa aacgaaaacg gaacggaaac gggatataca aaacggtaaa 1620
cggaaacgga aacggtagag ctagtttccc gaccgtttca ccgggatccc gtttttaatc 1680
gggatgatcc cgtttcgtta ccgtattttc taattcggga tgactgcaat atggccagct 1740
ccaactccca tccataacca ctgaggccca gcccatgtaa gaaataccta gcgaacgctg 1800
ctctgcctct ctcccaggcg gccaggcacc acacgagtaa cagcatcaca cattcacacg 1860
ccgccacgcg cccacgccgg agtccggacg ccgccagccg cacgccgacg ccggcgacgc 1920
gtctcgctct cgcctgctct ctccgactct ccctgtctcc cagccggccg gccgctgggc 1980
tgcaccaggc accacacgcg gtgacggccg tgacgcggca cgccggacgc agacgccgcc 2040
atccacggtc cgccctccac tccactgctc gcgactcgcc catccgcgcc gcggtccgcc 2100
catccgccca gacctccact ccactgctcg cccatccgcg gtccgcccat ccgccatccg 2160
ccatctgcgg tcagcggtcc tccagacctc cactgctcgg cgctcgccca tccggccatc 2220
cgcggtctcc ctgtctccac ggctgctcac aggctcacag cacttagcag tacagcacgt 2280
cagcaccatt gcaccaagct gttgtgtcat ttgtgtgctg tccaggggct ctgcaacacc 2340
tgctgattgc tgtccagccg tccaggtgct cacaagtcac agcagtacag caccaagctg 2400
attgctgaac acctgctgtc cagggctctg ctctccactt cggctagccg gctacgactc 2460
cattcctcag atgacgcctc cggttggaaa taatcctccc tcaggctcag ccataagatt 2520
ggccaagttg atgtctacca caagagcgcc ttctactcgc aaaacaaatt ccgtattctc 2580
tgcatatgct caaggtatat attagaaaaa cagtagcaat agcattagca ttactaattg 2640
gttgtagatt gggaagcatc atattgactg tagaataata cgaaaaatct gtttataaca 2700
gggttgaaaa gaaaagctga agcctcttct agtcggattc agaatgtacg tgcacgtgcg 2760
cgtgggcatg gatgtggccg cacatcacca tcatcatcaa cagctgaggc cgagaggcat 2820
tttattcaga gtgtaagcag tagtaatgca aatggtacag ctacagatcc gagtcaagat 2880
gatatggcta ttgttcatga accacaacca caaccacaac cacaaccaga accacaacca 2940
cagccacaac ctgaacccga agaagaagca ccacagaaga gggcaaagaa gtgcacatcg 3000
gatgtatggc agcatttcac caagaaggaa attgaagtgg aggtcgatgg aaagaaatac 3060
gttcaggtat ggggacattg caactttcct aattgcaagg ctaagtatag ggctgagggt 3120
catcatggaa caagcggatt tcgaaatcac ttgagaacat cacatagttt agttaaaggt 3180
cagttgtgtc taaaaagtga aaaggatcat ggcaaagaca taaatctcat tgagccttat 3240
aagtacgatg aagtggttag cctaaagaag cttgagctcg gtagcaattc ccgaggctgt 3300
agccgacgat ggtgcgccag gagagttgtt gattcagaat tccatcctac gatctagtaa 3360
catagatgac accgcgcgcg ataatttatc ctagtttgcg cgctatattt tgttttctat 3420
cgcgtattaa atgtataatt gcgggactct aatcataaaa acccatctca taaataacgt 3480
catgcattac atgttaatta ttacatgctt aacgtaattc aacagaaatt atatgataat 3540
catcgcaaga ccggcaacag gattcaatct taagaaactt tattgccaaa tgtttgaacg 3600
atcgagctct tagagttcgt cgtgtttgta tagttcatcc atgccatgtg taatcccagc 3660
agctgttaca aactcaagaa ggaccatgtg gtctctcttt tcgttgggat ctttcgaaag 3720
ggcggattgt gtggacaggt aatggttgtc cggtaaaagg acagggccat cgccaattgg 3780
agtattttgt tgataatgat cagcgagttg cacgccgccg tcttcgatgt tgtggcgggt 3840
cttgaagttg gctttgatgc cgttcttttg cttgtcggcc atgatgtata cgttgtggga 3900
gttgtagttg tattccaact tgtgaccgag gatgtttccg tcctccttga aatcgattcc 3960
cttaagctcg atcctgttga cgagggtgtc tccctcaaac ttgacttcag cacgtgtctt 4020
gtagttcccg tcgtccttga agaagatggt cctctcctgc acgtatccct caggcatggc 4080
gctcttgaag aagtcgtgcc gcttcatatg atctgggtat cttgaaaagc attgaacacc 4140
ataagagaaa gtagtgacaa gtgttggcca tggaacaggt agttttccag tagtgcaaat 4200
aaatttaagg gtaagttttc cgtatgttgc atcaccttca ccctctccac tgacagaaaa 4260
tttgtaccca ttaacatcac catctaattc aacaagaatt gggacaactc cagtgaaaag 4320
ttcttctcct ttactgaatt cggccgagga taatgatagg agaagtgaaa agatgagaaa 4380
gagaaaaaga ttagtcttca tacccgggga tcctctagag tcccccgtgt tctctccaaa 4440
tgaaatgaac ttccttatat agaggaaggg tcttgcgaag gatagtggga ttgtgcgtca 4500
tcccttacgt cagtggagat atcacatcaa tccacttgct ttgaagacgt ggttggaacg 4560
tcttcttttt ccacgatgct cctcgtgggt gggggtccat ctttgggacc actgtcggca 4620
gaggcatctt caacgatggc ctttccttta tcgcaatgat ggcatttgta ggagccacct 4680
tccttttcca ctatcttcac aataaagtga cagatagctg ggcaatggaa tccgaggagg 4740
tttccggata ttaccctttg ttgaaaagtc tcaactagtg gatccccaac atggtggagc 4800
acgacactct cgtctactcc aagaatatca aagatacagt ctcagaagac cagagggcta 4860
ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct cggattccat tgcccagcta 4920
tctgtcactt catcgaaagg acagtagaaa aggaagatgg cttctacaaa tgccatcatt 4980
gcgataaagg aaaggctatc gttcaagatg cctctaccga cagtggtccc aaagatggac 5040
ccccacccac gaggaacatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag 5100
tggattgatg tgatatctag atccccaaca tggtggagca cgacactctc gtctactcca 5160
agaatatcaa agatacagtc tcagaagacc agagggctat tgagactttt caacaaaggg 5220
taatatcggg aaacctcctc ggattccatt gcccagctat ctgtcacttc atcgaaagga 5280
cagtagaaaa ggaagatggc ttctacaaat gccatcattg cgataaagga aaggctatcg 5340
ttcaagatgc ctctaccgac agtggtccca aagatggacc cccacccacg aggaacatcg 5400
tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt gatatctaga 5460
tccccaacat ggtggagcac gacactctcg tctactccaa gaatatcaaa gatacagtct 5520
cagaagacca gagggctatt gagacttttc aacaaagggt aatatcggga aacctcctcg 5580
gattccattg cccagctatc tgtcacttca tcgaaaggac agtagaaaag gaagatggct 5640
tctacaaatg ccatcattgc gataaaggaa aggctatcgt tcaagatgcc tctaccgaca 5700
gtggtcccaa agatggaccc ccacccacga ggaacatcgt ggaaaaagaa gacgttccaa 5760
ccacgtcttc aaagcaagtg gattgatgtg atatctagat ccccaacatg gtggagcacg 5820
acactctcgt ctactccaag aatatcaaag atacagtctc agaagaccag agggctattg 5880
agacttttca acaaagggta atatcgggaa acctcctcgg attccattgc ccagctatct 5940
gtcacttcat cgaaaggaca gtagaaaagg aagatggctt ctacaaatgc catcattgcg 6000
ataaaggaaa ggctatcgtt caagatgcct ctaccgacag tggtcccaaa gatggacccc 6060
cacccacgag gaacatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg 6120
attgatgtga tatctaacta gtaatatatg ttttcatgtg tgattttacc gaacaaaaat 6180
accggttccc gtccgatttc gactttaacc cgaccggatc gtatcggttt tcgattaccg 6240
tatttatccc gttcgttttc gttaccggta tatcccgttt tcgtttccgt cccgcaagtt 6300
aaatatgaaa atgaaaacgg tagaggtatt ttaccgaccg ttaccgaccg ttttcatccc 6360
taaaggtgta tattgagaat gtggggtgtg agggtattat ggggaagtat gtagagtgta 6420
ttaagatagt ttttaattga aaaatcaaat gtgaggttta ttcaacaatt tgtgggatgt 6480
taatatacta agcctaaaaa aaacagaaaa tgatttcgaa gtatcagttc gtctctaatt 6540
cttacgaata ttatttatgg acttttctat tttcatattc tcatattcac attataaatt 6600
aggaaagaaa ttcacataat tgaaattttc acactataaa ctaggaaata aaacagaaaa 6660
ttgaagttgt atagatcatg ataaatgagg aaaattaaca gaaaattgaa gttcaccaag 6720
tactaagaac actggattac cacactggta ttgcaatttt taaataatca cgttgttttc 6780
cttgatataa tcctgaaaat ccatattata aaaagcctag gggtctaaga tgtgtctgca 6840
aatataaggc aattgagaaa cgcacaagag agaaaggtta tggcgacatt ggtgaggaag 6900
ttctttgtgc ttgttttgtt gctcatcatc tttctagctt ctggtaacta cgtactctca 6960
ttctctccct ctcacacaaa caattagtaa acatatatat tggaagaaat caaactcgaa 7020
acttattttc ttcgtaaatc attaggccac ggtcatccct attaattaac cccgagggct 7080
tgatttttaa cagttaaaat tacttaaatg atgagttagt attttctccc tcagcttatt 7140
ttgaagctca aaatttagtg aatgcgtgat atatcatctg tattaaatta ttaatcaatc 7200
tttttatata tcatttgtat tattcacatg cagatgagac agtgaagaat actgaagcta 7260
ttcaaatcgg ggactgtaca gaacttatcg gcggatgcac caggacacct ccctgcgtca 7320
gaaaatgcaa gaggcacggt tgcgatggcg gcaaatgctc cttattcctc tgtttgtgca 7380
acaagccttg ccctccatga aattatatta tttttatttc taaaataata gcccaccatg 7440
caaattgcaa ggcgctgcaa ttagtgt 7467
<210> 25
<211> 231
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 25
atggcgacat tggtgaggaa gttctttgtg cttgttttgt tgctcatcat ctttctagct 60
tctgatgaga cagtgaagaa tactgaagct attcaaatcg gggactgtac agaacttatc 120
ggcggatgca ccaggacacc tccctgcgtc agaaaatgca agaggcacgg ttgcgatggc 180
ggcaaatgct ccttattcct ctgtttgtgc aacaagcctt gccctccatg a 231
<210> 26
<211> 76
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 26
Met Ala Thr Leu Val Arg Lys Phe Phe Val Leu Val Leu Leu Leu Ile
1 5 10 15
Ile Phe Leu Ala Ser Asp Glu Thr Val Lys Asn Thr Glu Ala Ile Gln
20 25 30
Ile Gly Asp Cys Thr Glu Leu Ile Gly Gly Cys Thr Arg Thr Pro Pro
35 40 45
Cys Val Arg Lys Cys Lys Arg His Gly Cys Asp Gly Gly Lys Cys Ser
50 55 60
Leu Phe Leu Cys Leu Cys Asn Lys Pro Cys Pro Pro
65 70 75
<210> 28
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 28
acgatggact ccagcggccg cvnvnnngga a 31
<210> 28
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 28
acgatggact ccagcggccg cbnbnnnggt t 31
<210> 29
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 29
acgatggact ccagcggccg chnvnnncca c 31
<210> 30
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 30
acgatggact ccagcggccg cvvnvnnncc aa 32
<210> 31
<211> 16
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 31
acgatggact ccagag 16
<210> 32
<211> 50
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 32
gtctcaaata tatgttttca tgtgtgattt taccgaacaa aaataccggt 50
<210> 33
<211> 59
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 33
acgatggact ccagtccggc ccgtccgatt tcgactttaa cccgaccgga tcgtatcgg 59
<210> 34
<211> 45
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 34
cccgttcgtt ttcgttaccg gtatatcccg ttttcgtttc cgtcc 45
<210> 35
<211> 41
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 35
gagaacacgg gggactctag aatggcgaca ttggtgagga a 41
<210> 36
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 36
ttgaacgatc tcatggaggg caaggcttg 29
<210> 37
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 37
ccctccatga gatcgttcaa acatttggca ataa 34
<210> 38
<211> 47
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 38
agtcttcata cccggggatc ctgttctctc caaatgaaat gaacttc 47

Claims (2)

1.一种调控植物根系变短的小肽，其特征在于，所述小肽的氨基酸序列如SEQ IDNo.26所示。
2.根据权利要求1所述的调控植物根系变短的小肽，其特征在于，所述小肽的核苷酸序列如SEQ ID No.25所示。
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CN1137995C (zh) * 2001-05-21 2004-02-11 中国科学院遗传研究所 一种建立植物基因标签系统的方法
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CN103305541B (zh) * 2013-06-19 2016-02-10 西南大学 一种激活标签Ac/Ds转座系统及其在植物突变体库构建中的应用
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