pEGM-7ZF(+)

pEGM-7ZF(+)

pEGM-7ZF(+)载体基本信息

pEGM-7ZF(+)载体质粒图谱和多克隆位点信息

pEGM-7ZF(+)载体简介

The pGEM-7Zf(+) Vector(a) is a derivative of the pGEM?-3Zf(+) Vector(a) and contains the origin of replication of the filamentous phage f1. The plasmid serves as a standard cloning vector, as a template for in vitro transcription, and it can be used for the production of circular ssDNA. The plasmid contains SP6 and T7 RNA polymerase promoters flanking a region of multiple cloning sites within the α-peptide coding region of β-galactosidase (1). Insertional inactivation of the α-peptide allows recombinant clones to be directly identified by color screening on indicator plates. The multiple cloning region is unique and includes restriction sites for Apa I, Aat II, Sph I, Xba I, Xho I, EcoR I, Kpn I, Sma I, Csp45 I, Cla I, Hind III, BamH I, Sac I, BstX I and Nsi I. This arrangement is designed specifically for generation of unidirectional deletions with the Erase-a-Base? System(b). The polylinker contains restriction enzyme sites that produce 5′ overhangs or blunt ends (sensitive to Exonuclease III) flanked on both sides by blocks of restriction sites that generate 3′ overhangs (resistant to Exonuclease III). For induction of ssDNA, bacterial cells containing pGEM?-7Zf(+) recombinants are infected with an appropriate helper phage. The plasmid then enters the f1 replication mode, and the resulting ssDNA is exported from the cell as an encapsidated viruslike particle. The sequence of the ssDNA rescued upon infection with helper phage is complementary to the sequence shown in Figure 1. The exported ssDNA can be used for mutagenesis in vitro or can be sequenced using the T7 Promoter Primer or pUC/M13 Forward Primer.

pEGM-7ZF(+)载体序列

ORIGIN

    1 GGGCGAATTG GGCCCGACGT CGCATGCTCC TCTAGACTCG AGGAATTCGG TACCCCGGGT

   61 TCGAAATCGA TAAGCTTGGA TCCGGAGAGC TCCCAACGCG TTGGATGCAT AGCTTGAGTA

  121 TTCTATAGTG TCACCTAAAT AGCTTGGCGT AATCATGGTC ATAGCTGTTT CCTGTGTGAA

  181 ATTGTTATCC GCTCACAATT CCACACAACA TACGAGCCGG AAGCATAAAG TGTAAAGCCT

  241 GGGGTGCCTA ATGAGTGAGC TAACTCACAT TAATTGCGTT GCGCTCACTG CCCGCTTTCC

  301 AGTCGGGAAA CCTGTCGTGC CAGCTGCATT AATGAATCGG CCAACGCGCG GGGAGAGGCG

  361 GTTTGCGTAT TGGGCGCTCT TCCGCTTCCT CGCTCACTGA CTCGCTGCGC TCGGTCGTTC

  421 GGCTGCGGCG AGCGGTATCA GCTCACTCAA AGGCGGTAAT ACGGTTATCC ACAGAATCAG

  481 GGGATAACGC AGGAAAGAAC ATGTGAGCAA AAGGCCAGCA AAAGGCCAGG AACCGTAAAA

  541 AGGCCGCGTT GCTGGCGTTT TTCGATAGGC TCCGCCCCCC TGACGAGCAT CACAAAAATC

  601 GACGCTCAAG TCAGAGGTGG CGAAACCCGA CAGGACTATA AAGATACCAG GCGTTTCCCC

  661 CTGGAAGCTC CCTCGTGCGC TCTCCTGTTC CGACCCTGCC GCTTACCGGA TACCTGTCCG

  721 CCTTTCTCCC TTCGGGAAGC GTGGCGCTTT CTCATAGCTC ACGCTGTAGG TATCTCAGTT

  781 CGGTGTAGGT CGTTCGCTCC AAGCTGGGCT GTGTGCACGA ACCCCCCGTT CAGCCCGACC

  841 GCTGCGCCTT ATCCGGTAAC TATCGTCTTG AGTCCAACCC GGTAAGACAC GACTTATCGC

  901 CACTGGCAGC AGCCACTGGT AACAGGATTA GCAGAGCGAG GTATGTAGGC GGTGCTACAG

  961 AGTTCTTGAA GTGGTGGCCT AACTACGGCT ACACTAGAAG GACAGTATTT GGTATCTGCG

 1021 CTCTGCTGAA GCCAGTTACC TTCGGAAAAA GAGTTGGTAG CTCTTGATCC GGCAAACAAA

 1081 CCACCGCTGG TAGCGGTGGT TTTTTTGTTT GCAAGCAGCA GATTACGCGC AGAAAAAAAG

 1141 GATCTCAAGA AGATCCTTTG ATCTTTTCTA CGGGGTCTGA CGCTCAGTGG AACGAAAACT

 1201 CACGTTAAGG GATTTTGGTC ATGAGATTAT CAAAAAGGAT CTTCACCTAG ATCCTTTTAA

 1261 ATTAAAAATG AAGTTTTAAA TCAATCTAAA GTATATATGA GTAAACTTGG TCTGACAGTT

 1321 ACCAATGCTT AATCAGTGAG GCACCTATCT CAGCGATCTG TCTATTTCGT TCATCCATAG

 1381 TTGCCTGACT CCCCGTCGTG TAGATAACTA CGATACGGGA GGGCTTACCA TCTGGCCCCA

 1441 GTGCTGCAAT GATACCGCGA GACCCACGCT CACCGGCTCC AGATTTATCA GCAATAAACC

 1501 AGCCAGCCGG AAGGGCCGAG CGCAGAAGTG GTCCTGCAAC TTTATCCGCC TCCATCCAGT

 1561 CTATTAATTG TTGCCGGGAA GCTAGAGTAA GTAGTTCGCC AGTTAATAGT TTGCGCAACG

 1621 TTGTTGGCAT TGCTACAGGC ATCGTGGTGT CACGCTCGTC GTTTGGTATG GCTTCATTCA

 1681 GCTCCGGTTC CCAACGATCA AGGCGAGTTA CATGATCCCC CATGTTGTGC AAAAAAGCGG

 1741 TTAGCTCCTT CGGTCCTCCG ATCGTTGTCA GAAGTAAGTT GGCCGCAGTG TTATCACTCA

 1801 TGGTTATGGC AGCACTGCAT AATTCTCTTA CTGTCATGCC ATCCGTAAGA TGCTTTTCTG

 1861 TGACTGGTGA GTACTCAACC AAGTCATTCT GAGAATACCG CGCCCGGCGA CCGAGTTGCT

 1921 CTTGCCCGGC GTCAATACGG GATAATAGTG TATGACATAG CAGAACTTTA AAAGTGCTCA

 1981 TCATTGGAAA ACGTTCTTCG GGGCGAAAAC TCTCAAGGAT CTTACCGCTG TTGAGATCCA

 2041 GTTCGATGTA ACCCACTCGT GCACCCAACT GATCTTCAGC ATCTTTTACT TTCACCAGCG

 2101 TTTCTGGGTG AGCAAAAACA GGAAGGCAAA ATGCCGCAAA AAAGGGAATA AGGGCGACAC

 2161 GGAAATGTTG AATACTCATA CTCTTCCTTT TTCAATATTA TTGAAGCATT TATCAGGGTT

 2221 ATTGTCTCAT GAGCGGATAC ATATTTGAAT GTATTTAGAA AAATAAACAA ATAGGGGTTC

 2281 CGCGCACATT TCCCCGAAAA GTGCCACCTG TATGCGGTGT GAAATACCGC ACAGATGCGT

 2341 AAGGAGAAAA TACCGCATCA GGCGACGCGC CCTGTAGCGG CGCATTAAGC GCGGCGGGTG

 2401 TGGTGGTTAC GCGCAGCGTG ACCGCTACAC TTGCCAGCGC CCTAGCGCCC GCTCCTTTCG

 2461 CTTTCTTCCC TTCCTTTCTC GCCACGTTCG CCGGCTTTCC CCGTCAAGCT CTAAATCGGG

 2521 GGCTCCCTTT AGGGTTCCGA TTTAGAGCTT TACGGCACCT CGACCGCAAA AAACTTGATT

 2581 TGGGTGATGG TTCACGTAGT GGGCCATCGC CCTGATAGAC GGTTTTTCGC CCTTTGACGT

 2641 TGGAGTCCAC GTTCTTTAAT AGTGGACTCT TGTTCCAAAC TGGAACAACA CTCAACCCTA

 2701 TCTCGGTCTA TTCTTTTGAT TTATAAGGGA TTTTGCCGAT TTCGGCCTAT TGGTTAAAAA

 2761 ATGAGCTGAT TTAACAAATA TTTAACGCGA ATTTTAACAA AATATTAACG TTTACAATTT

 2821 CCATTCGCCA TTCAGGCTGC GCAACTGTTG GGAAGGGCGA TCGGTGCGGG CCTCTTCGCT

 2881 ATTACGCCAG CTGGCGAAAG GGGGATGTGC TGCAAGGCGA TTAAGTTGGG TAACGCCAGG

 2941 GTTTTCCCAG TCACGACGTT GTAAAACGAC GGCCAGTGAA TTGTAATACG ACTCACTATA

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