pFastBacHT A昆虫表达载体说明

pFastBacHTA编号  载体名称  北京华越洋生物KCVECT130  pFastBacHT  A  pFastBacHTA载体基本信息载体名称:pFastBacHTA,pFastBacHTA,pFastBacHTA,pFastBacHTA质粒类型:昆虫细胞表达载体；杆状病毒表达载体启动子:Polyhedrin表达水平:高克隆方法:多克隆位点，限制性内切酶载体大小:4856bp5'测序引物:pFastbac-F:TATTCCGGATTATTCATACC3'测序引物:pFastBac-R:ACAAATGTGGTATGGCTGA载体标签:N-Histag载体抗性:Ampicillin氨苄筛选标记:Gentamicin庆大霉素备注:含TEV切割位点稳定性:稳表达Stable组成型:组成型Constitutive病毒/非病毒:杆状病毒pFastBacHTA载体质粒图谱和多克隆位点信息      pFastBacHTA载体简介  Bac-­‐to-­‐BacHT  Vector  Kit  设计用于在大肠杆菌  (E.  coli)  中生成杆状病毒载体后，在  Sf9、Sf21  或  High  Five  细胞中表达和纯化带组氨酸标签的重组蛋白。  pFastBacHT  载体具    有以下特点：    ⇒用于表达蛋白质的多角体强启动子    ⇒用于简化克隆的三个读码框    ⇒用于轻松纯化重组融合蛋白的  N-­‐端  6xHis  标签    ⇒用于在蛋白质纯化后去除组氨酸标签的  TEV  蛋白酶切割位点    引用及参考文献    DEF6,  a  Novel  Substrate  for  the  Tec  Kinase  ITK,  Contains  a  Glutamine-­‐rich  Aggregation-­‐prone  Region  and  Forms  Cytoplasmic  Granules  that  Co-­‐localize  with  P-­‐bodies.  Hey  F,  Czyzewicz  N,  Jones  P,  Sablitzky  F,    J  Biol  Chem  (2012)  287:31073-­‐31084  Product  usage:  na    Cytoskeletal  changes  regulated  by  the  PAK4  serine/threonine  kinase  are  mediated  by  LIM  kinase  1  and  cofilin.  Dan  C;  Kelly  A;  Bernard  O;  Minden  A;    J  Biol  Chem  (2001)  276:32115-­‐32136  Product  usage:  pFASTBAC/HT  and  pFASTBAC-­‐1  vectors  were  used  to  transfect  with  Sf9  cells  with  Cellfectin.    Sp3  Represses  Gene  Expression  via  the  Titration  of  Promoter-­‐specific  Transcription  Factors.  Kennett  Sarah  B;  Moorefield  K  Scott;  Horowitz  Jonathan  M;    J  Biol  Chem  (2002)  277:9780-­‐9789    Product  usage:    Baculovirus  stocks  encoding  Sp1,  Sp3,  M1,  and  M2  were  prepared  using  the  PCR,  appropriate  primers,  and  pCMV4-­‐Sp1/flu,  pBSK-­‐Sp3/flu,  pCR-­‐M1/flu,  and  pCR-­‐M2/flu  as  substrates.  PCR  reactions  employed  pGEX-­‐M2  using  Taq  polymerase.  Amplified  cDNAs  were  subcloned  in  pFASTBacHTA.    pFastBacHTA载体序列  ORIGIN1GACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACC61GCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCC121ACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTT181AGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGG241CCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGT301GGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTA361TAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTT421AACGCGAATTTTAACAAAATATTAACGTTTACAATTTCAGGTGGCACTTTTCGGGGAAAT481GTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATG541AGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAA601CATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCAC661CCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTAC721ATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTT781CCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCC841GGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCA901CCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCC961ATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAG1021GAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAA1081CCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATG1141GCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAA1201TTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCG1261GCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATT1321GCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGT1381CAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAG1441CATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCAT1501TTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCT1561TAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCT1621TGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCA1681GCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTC1741AGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTC1801AAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCT1861GCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAG1921GCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACC1981TACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGG2041AGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAG2101CTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTT2161GAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAAC2221GCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCG2281TTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGC2341CGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCTGATG2401CGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCAGACCAGCCGCGTAACCT2461GGCAAAATCGGTTACGGTTGAGTAATAAATGGATGCCCTGCGTAAGCGGGTGTGGGCGGA2521CAATAAAGTCTTAAACTGAACAAAATAGATCTAAACTATGACAATAAAGTCTTAAACTAG2581ACAGAATAGTTGTAAACTGAAATCAGTCCAGTTATGCTGTGAAAAAGCATACTGGACTTT2641TGTTATGGCTAAAGCAAACTCTTCATTTTCTGAAGTGCAAATTGCCCGTCGTATTAAAGA2701GGGGCGTGGCCAAGGGCATGGTAAAGACTATATTCGCGGCGTTGTGACAATTTACCGAAC2761AACTCCGCGGCCGGGAAGCCGATCTCGGCTTGAACGAATTGTTAGGTGGCGGTACTTGGG2821TCGATATCAAAGTGCATCACTTCTTCCCGTATGCCCAACTTTGTATAGAGAGCCACTGCG2881GGATCGTCACCGTAATCTGCTTGCACGTAGATCA