pET-28b(+)

pET-28b(+)

pET28b载体基本信息

pET28b载体质粒图谱和多克隆位点信息

pET28b载体简介

pET-28b载体带有一个N端的His/Thrombin/T7蛋白标签，同时含有一个可以选择的C端His标签。pET28b载体的单一的 多克隆位点见上面的环状质粒图谱。注意：载体序列是以pBR322质粒的编码规矩进行编码的，所以T7蛋白表达区在质粒图谱上面是反向的。

T7 RNA聚合酶启动的克隆和表达区域在质粒图谱中也被标注了出来。质粒的F1复制子是被定向的，所以在T7噬菌体聚合酶的作用下，包含有蛋白编码序列的病毒 粒子能够产生，并启动蛋白表达，同时蛋白表达将被T7终止子序列(Cat. No. 69337-3)的作用下终止蛋白翻译。

The pET-28a-c(+) vectors carry an N-terminal His•Tag®/ thrombin/ T7•Tag® configuration plus an optional C-terminal His•Tag sequence. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circular map. The cloning/expression region of the coding strand transcribed by T7 RNA polymeraseis shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single stranded sequencing should be performed using the T7 terminator primer (Cat. No. 69337-3).

pET28b载体序列

ORIGIN

    1 ATCCGGATAT AGTTCCTCCT TTCAGCAAAA AACCCCTCAA GACCCGTTTA GAGGCCCCAA

   61 GGGGTTATGC TAGTTATTGC TCAGCGGTGG CAGCAGCCAA CTCAGCTTCC TTTCGGGCTT

  121 TGTTAGCAGC CGGATCTCAG TGGTGGTGGT GGTGGTGCTC GAGTGCGGCC GCAAGCTTGT

  181 CGACGGAGCT CGAATTCGGA TCCCGACCCA TTTGCTGTCC ACCAGTCATG CTAGCCATAT

  241 GGCTGCCGCG CGGCACCAGG CCGCTGCTGT GATGATGATG ATGATGGCTG CTGCCCATGG

  301 TATATCTCCT TCTTAAAGTT AAACAAAATT ATTTCTAGAG GGGAATTGTT ATCCGCTCAC

  361 AATTCCCCTA TAGTGAGTCG TATTAATTTC GCGGGATCGA GATCTCGATC CTCTACGCCG

  421 GACGCATCGT GGCCGGCATC ACCGGCGCCA CAGGTGCGGT TGCTGGCGCC TATATCGCCG

  481 ACATCACCGA TGGGGAAGAT CGGGCTCGCC ACTTCGGGCT CATGAGCGCT TGTTTCGGCG

  541 TGGGTATGGT GGCAGGCCCC GTGGCCGGGG GACTGTTGGG CGCCATCTCC TTGCATGCAC

  601 CATTCCTTGC GGCGGCGGTG CTCAACGGCC TCAACCTACT ACTGGGCTGC TTCCTAATGC

  661 AGGAGTCGCA TAAGGGAGAG CGTCGAGATC CCGGACACCA TCGAATGGCG CAAAACCTTT

  721 CGCGGTATGG CATGATAGCG CCCGGAAGAG AGTCAATTCA GGGTGGTGAA TGTGAAACCA

  781 GTAACGTTAT ACGATGTCGC AGAGTATGCC GGTGTCTCTT ATCAGACCGT TTCCCGCGTG

  841 GTGAACCAGG CCAGCCACGT TTCTGCGAAA ACGCGGGAAA AAGTGGAAGC GGCGATGGCG

  901 GAGCTGAATT ACATTCCCAA CCGCGTGGCA CAACAACTGG CGGGCAAACA GTCGTTGCTG

  961 ATTGGCGTTG CCACCTCCAG TCTGGCCCTG CACGCGCCGT CGCAAATTGT CGCGGCGATT

 1021 AAATCTCGCG CCGATCAACT GGGTGCCAGC GTGGTGGTGT CGATGGTAGA ACGAAGCGGC

 1081 GTCGAAGCCT GTAAAGCGGC GGTGCACAAT CTTCTCGCGC AACGCGTCAG TGGGCTGATC

 1141 ATTAACTATC CGCTGGATGA CCAGGATGCC ATTGCTGTGG AAGCTGCCTG CACTAATGTT

 1201 CCGGCGTTAT TTCTTGATGT CTCTGACCAG ACACCCATCA ACAGTATTAT TTTCTCCCAT

 1261 GAAGACGGTA CGCGACTGGG CGTGGAGCAT CTGGTCGCAT TGGGTCACCA GCAAATCGCG

 1321 CTGTTAGCGG GCCCATTAAG TTCTGTCTCG GCGCGTCTGC GTCTGGCTGG CTGGCATAAA

 1381 TATCTCACTC GCAATCAAAT TCAGCCGATA GCGGAACGGG AAGGCGACTG GAGTGCCATG

 1441 TCCGGTTTTC AACAAACCAT GCAAATGCTG AATGAGGGCA TCGTTCCCAC TGCGATGCTG

 1501 GTTGCCAACG ATCAGATGGC GCTGGGCGCA ATGCGCGCCA TTACCGAGTC CGGGCTGCGC

 1561 GTTGGTGCGG ATATCTCGGT AGTGGGATAC GACGATACCG AAGACAGCTC ATGTTATATC

 1621 CCGCCGTTAA CCACCATCAA ACAGGATTTT CGCCTGCTGG GGCAAACCAG CGTGGACCGC

 1681 TTGCTGCAAC TCTCTCAGGG CCAGGCGGTG AAGGGCAATC AGCTGTTGCC CGTCTCACTG

 1741 GTGAAAAGAA AAACCACCCT GGCGCCCAAT ACGCAAACCG CCTCTCCCCG CGCGTTGGCC

 1801 GATTCATTAA TGCAGCTGGC ACGACAGGTT TCCCGACTGG AAAGCGGGCA GTGAGCGCAA

 1861 CGCAATTAAT GTAAGTTAGC TCACTCATTA GGCACCGGGA TCTCGACCGA TGCCCTTGAG

 1921 AGCCTTCAAC CCAGTCAGCT CCTTCCGGTG GGCGCGGGGC ATGACTATCG TCGCCGCACT

 1981 TATGACTGTC TTCTTTATCA TGCAACTCGT AGGACAGGTG CCGGCAGCGC TCTGGGTCAT

 2041 TTTCGGCGAG GACCGCTTTC GCTGGAGCGC GACGATGATC GGCCTGTCGC TTGCGGTATT

 2101 CGGAATCTTG CACGCCCTCG CTCAAGCCTT CGTCACTGGT CCCGCCACCA AACGTTTCGG

 2161 CGAGAAGCAG GCCATTATCG CCGGCATGGC GGCCCCACGG GTGCGCATGA TCGTGCTCCT

 2221 GTCGTTGAGG ACCCGGCTAG GCTGGCGGGG TTGCCTTACT GGTTAGCAGA ATGAATCACC

 2281 GATACGCGAG CGAACGTGAA GCGACTGCTG CTGCAAAACG TCTGCGACCT GAGCAACAAC

 2341 ATGAATGGTC TTCGGTTTCC GTGTTTCGTA AAGTCTGGAA ACGCGGAAGT CAGCGCCCTG

 2401 CACCATTATG TTCCGGATCT GCATCGCAGG ATGCTGCTGG CTACCCTGTG GAACACCTAC

 2461 ATCTGTATTA ACGAAGCGCT GGCATTGACC CTGAGTGATT TTTCTCTGGT CCCGCCGCAT

 2521 CCATACCGCC AGTTGTTTAC CCTCACAACG TTCCAGTAAC CGGGCATGTT CATCATCAGT

 2581 AACCCGTATC GTGAGCATCC TCTCTCGTTT CATCGGTATC ATTACCCCCA TGAACAGAAA

 2641 TCCCCCTTAC ACGGAGGCAT CAGTGACCAA ACAGGAAAAA ACCGCCCTTA ACATGGCCCG

 2701 CTTTATCAGA AGCCAGACAT TAACGCTTCT GGAGAAACTC AACGAGCTGG ACGCGGATGA

 2761 ACAGGCAGAC ATCTGTGAAT CGCTTCACGA CCACGCTGAT GAGCTTTACC GCAGCTGCCT

 2821 CGCGCGTTTC GGTGATGACG GTGAAAACCT CTGACACATG CAGCTCCCGG AGACGGTCAC

 2881 AGCTTGTCTG TAAGCGGATG CCGGGAGCAG ACAAGCCCGT CAGGGCGCGT CAGCGGGTGT

 2941 TGGCGGGTGT CGGGGCGCAG CCATGACCCA GTCACGTAGC GATAGCGGAG TGTATACTGG

 3001 CTTAACTATG CGGCATCAGA GCAGATTGTA CTGAGAGTGC ACCATATATG CGGTGTGAAA

 3061 TACCGCACAG ATGCGTAAGG AGAAAATACC GCATCAGGCG CTCTTCCGCT TCCTCGCTCA

 3121 CTGACTCGCT GCGCTCGGTC GTTCGGCTGC GGCGAGCGGT ATCAGCTCAC TCAAAGGCGG

 3181 TAATACGGTT ATCCACAGAA TCAGGGGATA ACGCAGGAAA GAACATGTGA GCAAAAGGCC

 3241 AGCAAAAGGC CAGGAACCGT AAAAAGGCCG CGTTGCTGGC GTTTTTCCAT AGGCTCCGCC

 3301 CCCCTGACGA GCATCACAAA AATCGACGCT CAAGTCAGAG GTGGCGAAAC CCGACAGGAC

 3361 TATAAAGATA CCAGGCGTTT CCCCCTGGAA GCTCCCTCGT GCGCTCTCCT GTTCCGACCC

 3421 TGCCGCTTAC CGGATACCTG TCCGCCTTTC TCCCTTCGGG AAGCGTGGCG CTTTCTCATA

 3481 GCTCACGCTG TAGGTATCTC AGTTCGGTGT AGGTCGTTCG CTCCAAGCTG GGCTGTGTGC

 3541 ACGAACCCCC CGTTCAGCCC GACCGCTGCG CCTTATCCGG TAACTATCGT CTTGAGTCCA

 3601 ACCCGGTAAG ACACGACTTA TCGCCACTGG CAGCAGCCAC TGGTAACAGG ATTAGCAGAG

 3661 CGAGGTATGT AGGCGGTGCT ACAGAGTTCT TGAAGTGGTG GCCTAACTAC GGCTACACTA

 3721 GAAGGACAGT ATTTGGTATC TGCGCTCTGC TGAAGCCAGT TACCTTCGGA AAAAGAGTTG

 3781 GTAGCTCTTG ATCCGGCAAA CAAACCACCG CTGGTAGCGG TGGTTTTTTT GTTTGCAAGC

 3841 AGCAGATTAC GCGCAGAAAA AAAGGATCTC AAGAAGATCC TTTGATCTTT TCTACGGGGT

 3901 CTGACGCTCA GTGGAACGAA AACTCACGTT AAGGGATTTT GGTCATGAAC AATAAAACTG

 3961 TCTGCTTACA TAAACAGTAA TACAAGGGGT GTTATGAGCC ATATTCAACG GGAAACGTCT

 4021 TGCTCTAGGC CGCGATTAAA TTCCAACATG GATGCTGATT TATATGGGTA TAAATGGGCT

 4081 CGCGATAATG TCGGGCAATC AGGTGCGACA ATCTATCGAT TGTATGGGAA GCCCGATGCG

 4141 CCAGAGTTGT TTCTGAAACA TGGCAAAGGT AGCGTTGCCA ATGATGTTAC AGATGAGATG

 4201 GTCAGACTAA ACTGGCTGAC GGAATTTATG CCTCTTCCGA CCATCAAGCA TTTTATCCGT

 4261 ACTCCTGATG ATGCATGGTT ACTCACCACT GCGATCCCCG GGAAAACAGC ATTCCAGGTA

 4321 TTAGAAGAAT ATCCTGATTC AGGTGAAAAT ATTGTTGATG CGCTGGCAGT GTTCCTGCGC

 4381 CGGTTGCATT CGATTCCTGT TTGTAATTGT CCTTTTAACA GCGATCGCGT ATTTCGTCTC

 4441 GCTCAGGCGC AATCACGAAT GAATAACGGT TTGGTTGATG CGAGTGATTT TGATGACGAG

 4501 CGTAATGGCT GGCCTGTTGA ACAAGTCTGG AAAGAAATGC ATAAACTTTT GCCATTCTCA

 4561 CCGGATTCAG TCGTCACTCA TGGTGATTTC TCACTTGATA ACCTTATTTT TGACGAGGGG

 4621 AAATTAATAG GTTGTATTGA TGTTGGACGA GTCGGAATCG CAGACCGATA CCAGGATCTT

 4681 GCCATCCTAT GGAACTGCCT CGGTGAGTTT TCTCCTTCAT TACAGAAACG GCTTTTTCAA

 4741 AAATATGGTA TTGATAATCC TGATATGAAT AAATTGCAGT TTCATTTGAT GCTCGATGAG

 4801 TTTTTCTAAG AATTAATTCA TGAGCGGATA CATATTTGAA TGTATTTAGA AAAATAAACA

 4861 AATAGGGGTT CCGCGCACAT TTCCCCGAAA AGTGCCACCT GAAATTGTAA ACGTTAATAT

 4921 TTTGTTAAAA TTCGCGTTAA ATTTTTGTTA AATCAGCTCA TTTTTTAACC AATAGGCCGA

 4981 AATCGGCAAA ATCCCTTATA AATCAAAAGA ATAGACCGAG ATAGGGTTGA GTGTTGTTCC

 5041 AGTTTGGAAC AAGAGTCCAC TATTAAAGAA CGTGGACTCC AACGTCAAAG GGCGAAAAAC

 5101 CGTCTATCAG GGCGATGGCC CACTACGTGA ACCATCACCC TAATCAAGTT TTTTGGGGTC

 5161 GAGGTGCCGT AAAGCACTAA ATCGGAACCC TAAAGGGAGC CCCCGATTTA GAGCTTGACG

 5221 GGGAAAGCCG GCGAACGTGG CGAGAAAGGA AGGGAAGAAA GCGAAAGGAG CGGGCGCTAG

 5281 GGCGCTGGCA AGTGTAGCGG TCACGCTGCG CGTAACCACC ACACCCGCCG CGCTTAATGC

 5341 GCCGCTACAG GGCGCGTCCC ATTCGCCA

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pET-28b(+)其他大肠杆菌表达载体：