pRSFDuet-1大肠杆菌表达载体说明

pRSFDuet-1  编号  名称  北京华越洋VECT-­‐530  pRSFDuet-­‐1    pRSFDuet1载体基本信息  别名:  pRSFDuet1,  pRSFDuet  1  质粒类型:  大肠杆菌蛋白双表达载体，有两个多克隆位点  表达水平:  高  克隆方法:  多克隆位点，限制性内切酶  载体大小:  3829  bp  5'  测序引物:  T7  5'  测序引物序列:      T7:  5'-­‐TAATACGACTCACTATAGGG-­‐3';  3'  测序引物:  T7t  3'  测序引物序列:      T7t:  5'-­‐TGCTAGTTATTGCTCAGCGG-­‐3'  载体标签:  N-­‐His,  C-­‐S  载体抗性:  Kanamycin  备注:  pRSFDuet-­‐1  carries  the  RSF1030  replicon,  lacI  gene  and  kanamycin  resistance  gene.  稳定性:  瞬时表达  Transient  组成型:  组成型  Constitutive  病毒/非病毒:  非病毒    pRSFDuet1载体质粒图谱和多克隆位点信息      pRSFDuet1载体简介  pRSFDuet-­‐1  is  designed  for  the  coexpression  of  two  target  ORFs.  The  vector  contains  two  multiple  cloning  sites  (MCS),  each  of  which  is  preceded  by  a  T7lac  promoter  and  ribosome  binding  site  (rbs).  The  vector  also  carries  the  RSF1030-­‐derived  RSF  replicon,  lacI  gene  and  kanamycin  resistance  gene  (KnR).  This  vector  can  be  used  in  combination  with  pACYCDuet™-­‐1  ,  pCDFDuet™-­‐1,  and  pETDuet™-­‐1  in  an  appropriate  host  strain  for  the  coexpression  of  4  to  8  target  proteins.  ORFs  inserted  into  MCS1  can  be  sequenced  using  the  ACYCDuetUP1  Primer  and  DuetDOWN1  Primer.  ORFs  inserted  into  MCS2  can  be  sequenced  using  the  DuetUP2  Primer  and  T7  Terminator  Primer.  Unique  sites  are  shown  on  the  circle  map.    pRSFDuet1载体序列  ORIGIN1GGGGAATTGTGAGCGGATAACAATTCCCCTGTAGAAATAATTTTGTTTAACTTTAATAAG61GAGATATACCATGGGCAGCAGCCATCACCATCATCACCACAGCCAGGATCCGAATTCGAG121CTCGGCGCGCCTGCAGGTCGACAAGCTTGCGGCCGCATAATGCTTAAGTCGAACAGAAAG181TAATCGTATTGTACACGGCCGCATAATCGAAATTAATACGACTCACTATAGGGGAATTGT241GAGCGGATAACAATTCCCCATCTTAGTATATTAGTTAAGTATAAGAAGGAGATATACATA301TGGCAGATCTCAATTGGATATCGGCCGGCCACGCGATCGCTGACGTCGGTACCCTCGAGT361CTGGTAAAGAAACCGCTGCTGCGAAATTTGAACGCCAGCACATGGACTCGTCTACTAGCG421CAGCTTAATTAACCTAGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGG481CCTCTAAACGGGTCTTGAGGGGTTTTTTGCTGAAACCTCAGGCATTTGAGAAGCACACGG541TCACACTGCTTCCGGTAGTCAATAAACCGGTAAACCAGCAATAGACATAAGCGGCTATTT601AACGACCCTGCCCTGAACCGACGACAAGCTGACGACCGGGTCTCCGCAAGTGGCACTTTT661CGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTAT721CCGCTCATGAATTAATTCTTAGAAAAACTCATCGAGCATCAAATGAAACTGCAATTTATT781CATATCAGGATTATCAATACCATATTTTTGAAAAAGCCGTTTCTGTAATGAAGGAGAAAA841CTCACCGAGGCAGTTCCATAGGATGGCAAGATCCTGGTATCGGTCTGCGATTCCGACTCG901TCCAACATCAATACAACCTATTAATTTCCCCTCGTCAAAAATAAGGTTATCAAGTGAGAA961ATCACCATGAGTGACGACTGAATCCGGTGAGAATGGCAAAAGTTTATGCATTTCTTTCCA1021GACTTGTTCAACAGGCCAGCCATTACGCTCGTCATCAAAATCACTCGCATCAACCAAACC1081GTTATTCATTCGTGATTGCGCCTGAGCGAGACGAAATACGCGGTCGCTGTTAAAAGGACA1141ATTACAAACAGGAATCGAATGCAACCGGCGCAGGAACACTGCCAGCGCATCAACAATATT1201TTCACCTGAATCAGGATATTCTTCTAATACCTGGAATGCTGTTTTCCCGGGGATCGCAGT1261GGTGAGTAACCATGCATCATCAGGAGTACGGATAAAATGCTTGATGGTCGGAAGAGGCAT1321AAATTCCGTCAGCCAGTTTAGTCTGACCATCTCATCTGTAACATCATTGGCAACGCTACC1381TTTGCCATGTTTCAGAAACAACTCTGGCGCATCGGGCTTCCCATACAATCGATAGATTGT1441CGCACCTGATTGCCCGACATTATCGCGAGCCCATTTATACCCATATAAATCAGCATCCAT1501GTTGGAATTTAATCGCGGCCTAGAGCAAGACGTTTCCCGTTGAATATGGCTCATACTCTT1561CCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATT1621TGAATGTATTTAGAAAAATAAACAAATAGGCATGCAGCGCTCTTCCGCTTCCTCGCTCAC1681TGACTCGCTACGCTCGGTCGTTCGACTGCGGCGAGCGGTGTCAGCTCACTCAAAAGCGGT1741AATACGGTTATCCACAGAATCAGGGGATAAAGCCGGAAAGAACATGTGAGCAAAAAGCAA1801AGCACCGGAAGAAGCCAACGCCGCAGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGC1861ATCACAAAAATCGACGCTCAAGCCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACC1921AGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCG1981GATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTT2041GGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCG2101TTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGAC2161ACGACTTATCGCCACTGGCAGCAGCCATTGGTAACTGATTTAGAGGACTTTGTCTTGAAG2221TTATGCACCTGTTAAGGCTAAACTGAAAGAACAGATTTTGGTGAGTGCGGTCCTCCAACC2281CACTTACCTTGGTTCAAAGAGTTGGTAGCTCAGCGAACCTTGAGAAAACCACCGTTGGTA2341GCGGTGGTTTTTCTTTATTTATGAGATGATGAATCAATCGGTCTATCAAGTCAACGAACA2401GCTATTCCGTTACTCTAGATTTCAGTGCAATTTATCTCTTCAAATGTAGCACCTGAAGTC2461AGCCCCATACGATATAAGTTGTAATTCTCATGTTAGTCATGCCCCGCGCCCACCGGAAGG2521AGCTGACTGGGTTGAAGGCTCTCAAGGGCATCGGTCGAGATCCCGGTGCCTAATGAGTGA2581GCTAACTTACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGT2641GCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCC2701AGGGTGGTTTTTCTTTTCACCAGTGAGACGGGCAACAGCTGATTGCCCTTCACCGCCTGG2761CCCTGAGAGAGTTGCAGCAAGCGGTCCACGCTGGTTTGCCCCAGCAGGCGAAAATCCTGT2821TTGATGGTGGTTAACGGCGGGATATAACATGAGCTGTCTTCGGTATCGTCGTATCCCACT2881ACCGAGATGTCCGCACCAACGCGCAGCCCGGACTCGGTAATGGCGCGCATTGCGCCCAGC2941GCCATCTGATCGTTGGCAACCAGCATCGCAGTGGGAACGATGCCCTCATTCAGCATTTGC3001ATGGTTTGTTGAAAACCGGACATGGCACTCCAGTCGCCTTCCCGTTCCGCTATCGGCTGA3061ATTTGATTGCGAGTGAGATATTTATGCCAGCCAGCCAGACGCAGACGCGCCGAGACAGAA3121CTTAATGGGCCCGCTAACAGCGCGATTTGCTGGTGACCCAATGCGACCAGATGCTCCACG3181CCCAGTCGCGTACCGTCTTCATGGGAGAAAATAATACTGTTGATGGGTGTCTGGTCAGAG3241ACATCAAGAAATAACGCCGGAACATTAGTGCAGGCAGCTTCCACAGCAATGGCATCCTGG3301TCATCCAGCGGATAGTTAATGATCAGCCCACTGACGCGTTGCGCGAGAAGATTGTGCACC3361GCCGCTTTACAGGCTTCGACGCCGCTT