pET-48b(+)

pET-48b(+)

pET48b载体基本信息

pET48b载体质粒图谱和多克隆位点信息

pET48b载体简介

pET-48b载体含有N端Trx和His标签，在标签后面紧跟着的是HRV 3C蛋白酶切位点。HRV 3C蛋白酶能够高特异性的识别LEVLFQ↓GP蛋白序列，能够在低温下高效切割掉融合标签序列。pET-48b载体还含有一个可选择的C端Thrombin蛋白酶切位点，紧接着位点后是S标签。

pET48b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。载体的单链测序应该使用S 18引物(Cat. No. 71262-3)。

The pET-48b(+) vector carries N-terminal Trx•Tag™ and His•Tag? coding sequences followed by a recognition site for the human rhinovirus (HRV) 3C protease. This protease is highly specifi c for cleavage of the sequence LEVLFQ↓GP (1), and is active at low temperatures (2). pET-48b(+) also contains an optional C-terminal thrombin recognition site followed by an S•Tag™ coding sequence. Unique restriction 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 circle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with the helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-stranded sequencing should be performed using the AS S•Tag 18mer Primer (Cat. No. 71262-3).

pET48b载体序列

ORIGIN

        1 ATCCGGATAT AGTTCCTCCT TTCAGCAAAA AACCCCTCAA GACCCGTTTA GAGGCCCCAA

       61 GGGGTTATGC TAGTTATTGC TCAGCGGTGG CAGCAGCCAA CTCAGCTTCC TTTCGGGCTT

      121 TGTTTAGCAG CCTAGGTTAA TTAAGCCTCG AGAGCAGCAG AAGTAGAGCT GTCCATGTGC

      181 TGGCGTTCGA ATTTAGCAGC AGCGGTTTCT TTACTACCGC GTGGCACCAG AGCGAGCTCT

      241 GCGGCCGCAA GCTTGTCGAC GGACGTCGGG CGCGCCAAGG CCTGTACAGA ATTCGGATCC

      301 TGGTACCCGG GTCCCTGAAA GAGGACTTCA AGAGCCGCGG AGTGATGGTG ATGGTGATGA

      361 CCAGAACCAC TAGATGGAGT AGGAGTAGGA GGATTGTTAT TAGAACCGCC ACCACCACTA

      421 GTATGGCCAG AACCAGAACC GGCCAGGTTA GCGTCGAGGA ACTCTTTCAA CTGACCTTTA

      481 GACAGTGCAC CCACTTTGGT TGCCGCCACT TCACCGTTTT TGAACAGCAG CAGAGTCGGG

      541 ATACCACGGA TGCCATATTT CGGCGCAGTG CCAGGGTTTT GATCGATGTT CAGTTTTGCA

      601 ACGGTCAGTT TGCCCTGATA TTCGTCAGCG ATTTCATCCA GAATCGGGGC GATCATTTTG

      661 CACGGACCGC ACCACTCTGC CCAGAAATCG ACGAGGATCG CCCCGTCCGC TTTGAGTACA

      721 TCCGTGTCAA AACTGTCGTC AGTCAGGTGA ATAATTTTAT CGCTCATATG TATATCTCCT

      781 TCTTAAAGTT AAACAAAATT ATTTCTAGAG GGGAATTGTT ATCCGCTCAC AATTCCCCTA

      841 TAGTGAGTCG TATTAATTTC GCGGGATCGA GATCGATCTC GATCCTCTAC GCCGGACGCA

      901 TCGTGGCCGG CATCACCGGC GCCACAGGTG CGGTTGCTGG CGCCTATATC GCCGACATCA

      961 CCGATGGGGA AGATCGGGCT CGCCACTTCG GGCTCATGAG CGCTTGTTTC GGCGTGGGTA

     1021 TGGTGGCAGG CCCCGTGGCC GGGGGACTGT TGGGCGCCAT CTCCTTGCAT GCACCATTCC

     1081 TTGCGGCGGC GGTGCTCAAC GGCCTCAACC TACTACTGGG CTGCTTCCTA ATGCAGGAGT

     1141 CGCATAAGGG AGAGCGTCGA GATCCCGGAC ACCATCGAAT GGCGCAAAAC CTTTCGCGGT

     1201 ATGGCATGAT AGCGCCCGGA AGAGAGTCAA TTCAGGGTGG TGAATGTGAA ACCAGTAACG

     1261 TTATACGATG TCGCAGAGTA TGCCGGTGTC TCTTATCAGA CCGTTTCCCG CGTGGTGAAC

     1321 CAGGCCAGCC ACGTTTCTGC GAAAACGCGG GAAAAAGTGG AAGCGGCGAT GGCGGAGCTG

     1381 AATTACATTC CCAACCGCGT GGCACAACAA CTGGCGGGCA AACAGTCGTT GCTGATTGGC

     1441 GTTGCCACCT CCAGTCTGGC CCTGCACGCG CCGTCGCAAA TTGTCGCGGC GATTAAATCT

     1501 CGCGCCGATC AACTGGGTGC CAGCGTGGTG GTGTCGATGG TAGAACGAAG CGGCGTCGAA

     1561 GCCTGTAAAG CGGCGGTGCA CAATCTTCTC GCGCAACGCG TCAGTGGGCT GATCATTAAC

     1621 TATCCGCTGG ATGACCAGGA TGCCATTGCT GTGGAAGCTG CCTGCACTAA TGTTCCGGCG

     1681 TTATTTCTTG ATGTCTCTGA CCAGACACCC ATCAACAGTA TTATTTTCTC CCATGAAGAC

     1741 GGTACGCGAC TGGGCGTGGA GCATCTGGTC GCATTGGGTC ACCAGCAAAT CGCGCTGTTA

     1801 GCGGGCCCAT TAAGTTCTGT CTCGGCGCGT CTGCGTCTGG CTGGCTGGCA TAAATATCTC

     1861 ACTCGCAATC AAATTCAGCC GATAGCGGAA CGGGAAGGCG ACTGGAGTGC CATGTCCGGT

     1921 TTTCAACAAA CCATGCAAAT GCTGAATGAG GGCATCGTTC CCACTGCGAT GCTGGTTGCC

     1981 AACGATCAGA TGGCGCTGGG CGCAATGCGC GCCATTACCG AGTCCGGGCT GCGCGTTGGT

     2041 GCGGACATCT CGGTAGTGGG ATACGACGAT ACCGAAGACA GCTCATGTTA TATCCCGCCG

     2101 TTAACCACCA TCAAACAGGA TTTTCGCCTG CTGGGGCAAA CCAGCGTGGA CCGCTTGCTG

     2161 CAACTCTCTC AGGGCCAGGC GGTGAAGGGC AATCAGCTGT TGCCCGTCTC ACTGGTGAAA

     2221 AGAAAAACCA CCCTGGCGCC CAATACGCAA ACCGCCTCTC CCCGCGCGTT GGCCGATTCA

     2281 TTAATGCAGC TGGCACGACA GGTTTCCCGA CTGGAAAGCG GGCAGTGAGC GCAACGCAAT

     2341 TAATGTAAGT TAGCTCACTC ATTAGGCACC GGGATCTCGA CCGATGCCCT TGAGAGCCTT

     2401 CAACCCAGTC AGCTCCTTCC GGTGGGCGCG GGGCATGACT AGCATGATCG TGCTCCTGTC

     2461 GTTGAGGACC CGGCTAGGCT GGCGGGGTTG CCTTACTGGT TAGCAGAATG AATCACCGAT

     2521 ACGCGAGCGA ACGTGAAGCG ACTGCTGCTG CAAAACGTCT GCGACCTGAG CAACAACATG

     2581 AATGGTCTTC GGTTTCCGTG TTTCGTAAAG TCTGGAAACG CGGAAGTCAG CGCCCTGCAC

     2641 CATTATGTTC CGGATCTGCA TCGCAGGATG CTGCTGGCTA CCCTGTGGAA CACCTACATC

     2701 TGTATTAACG AAGCGCTGGC ATTGACCCTG AGTGATTTTT CTCTGGTCCC GCCGCATCCA

     2761 TACCGCCAGT TGTTTACCCT CACAACGTTC CAGTAACCGG GCATGTTCAT CATCAGTAAC

     2821 CCGTATCGTG AGCATCCTCT CTCGTTTCAT CGGTATCATT ACCCCCATGA ACAGAAATCC

     2881 CCCTTACACG GAGGCATCAG TGACCAAACA GGAAAAAACC GCCCTTAACA TGGCCCGCTT

     2941 TATCAGAAGC CAGACATTAA CGCTTCTGGA GAAACTCAAC GAGCTGGACG CGGATGAACA

     3001 GGCAGACATC TGTGAATCGC TTCACGACCA CGCTGATGAG CTTTACCGCA GCTGCCTCGC

     3061 GCGTTTCGGT GATGACGGTG AAAACCTCTG ACACATGCAG CTCCCGGAGA CGGTCACAGC

     3121 TTGTCTGTAA GCGGATGCCG GGAGCAGACA AGCCCGTCAG GGCGCGTCAG CGGGTGTTGG

     3181 CGGGTGTCGG GGCGCAGCCA TGACCCAGTC ACGTAGCGAT AGCGGAGTGT ATACTGGCTT

     3241 AACTATGCGG CATCAGAGCA GATTGTACTG AGAGTGCACC ATATATGCGG TGTGAAATAC

     3301 CGCACAGATG CGTAAGGAGA AAATACCGCA TCAGGCGCTC TTCCGCTTCC TCGCTCACTG

     3361 ACTCGCTGCG CTCGGTCGTT CGGCTGCGGC GAGCGGTATC AGCTCACTCA AAGGCGGTAA

     3421 TACGGTTATC CACAGAATCA GGGGATAACG CAGGAAAGAA CATGTGAGCA AAAGGCCAGC

     3481 AAAAGGCCAG GAACCGTAAA AAGGCCGCGT TGCTGGCGTT TTTCCATAGG CTCCGCCCCC

     3541 CTGACGAGCA TCACAAAAAT CGACGCTCAA GTCAGAGGTG GCGAAACCCG ACAGGACTAT

     3601 AAAGATACCA GGCGTTTCCC CCTGGAAGCT CCCTCGTGCG CTCTCCTGTT CCGACCCTGC

     3661 CGCTTACCGG ATACCTGTCC GCCTTTCTCC CTTCGGGAAG CGTGGCGCTT TCTCATAGCT

     3721 CACGCTGTAG GTATCTCAGT TCGGTGTAGG TCGTTCGCTC CAAGCTGGGC TGTGTGCACG

     3781 AACCCCCCGT TCAGCCCGAC CGCTGCGCCT TATCCGGTAA CTATCGTCTT GAGTCCAACC

     3841 CGGTAAGACA CGACTTATCG CCACTGGCAG CAGCCACTGG TAACAGGATT AGCAGAGCGA

     3901 GGTATGTAGG CGGTGCTACA GAGTTCTTGA AGTGGTGGCC TAACTACGGC TACACTAGAA

     3961 GGACAGTATT TGGTATCTGC GCTCTGCTGA AGCCAGTTAC CTTCGGAAAA AGAGTTGGTA

     4021 GCTCTTGATC CGGCAAACAA ACCACCGCTG GTAGCGGTGG TTTTTTTGTT TGCAAGCAGC

     4081 AGATTACGCG CAGAAAAAAA GGATCTCAAG AAGATCCTTT GATCTTTTCT ACGGGGTCTG

     4141 ACGCTCAGTG GAACGAAAAC TCACGTTAAG GGATTTTGGT CATGAACAAT AAAACTGTCT

     4201 GCTTACATAA ACAGTAATAC AAGGGGTGTT ATGAGCCATA TTCAACGGGA AACGTCTTGC

     4261 TCTAGGCCGC GATTAAATTC CAACATGGAT GCTGATTTAT ATGGGTATAA ATGGGCTCGC

     4321 GATAATGTCG GGCAATCAGG TGCGACAATC TATCGATTGT ATGGGAAGCC CGATGCGCCA

     4381 GAGTTGTTTC TGAAACATGG CAAAGGTAGC GTTGCCAATG ATGTTACAGA TGAGATGGTC

     4441 AGACTAAACT GGCTGACGGA ATTTATGCCT CTTCCGACCA TCAAGCATTT TATCCGTACT

     4501 CCTGATGATG CATGGTTACT CACCACTGCG ATCCCCGGCA AAACAGCATT CCAGGTATTA

     4561 GAAGAATATC CTGATTCAGG TGAAAATATT GTTGATGCGC TGGCAGTGTT CCTGCGCCGG

     4621 TTGCATTCGA TTCCTGTTTG TAATTGTCCT TTTAACAGTG ATCGCGTATT TCGTCTCGCT

     4681 CAGGCGCAAT CACGAATGAA TAACGGTTTG GTTGATGCGA GTGATTTTGA TGACGAGCGT

     4741 AATGGCTGGC CTGTTGAACA AGTCTGGAAA GAAATGCATA AACTTTTGCC ATTCTCACCG

     4801 GATTCAGTCG TCACTCATGG TGATTTCTCA CTTGATAACC TTATTTTTGA CGAGGGGAAA

     4861 TTAATAGGTT GTATTGATGT TGGACGAGTC GGAATCGCAG ACCGATACCA GGATCTTGCC

     4921 ATCCTATGGA ACTGCCTCGG TGAGTTTTCT CCTTCATTAC AGAAACGGCT TTTTCAAAAA

     4981 TATGGTATTG ATAATCCTGA TATGAATAAA TTGCAGTTTC ATTTGATGCT CGATGAGTTT

     5041 TTCTAAGAAT TAATTCATGA GCGGATACAT ATTTGAATGT ATTTAGAAAA ATAAACAAAT

     5101 AGGGGTTCCG CGCACATTTC CCCGAAAAGT GCCACCTGAA ATTGTAAACG TTAATATTTT

     5161 GTTAAAATTC GCGTTAAATT TTTGTTAAAT CAGCTCATTT TTTAACCAAT AGGCCGAAAT

     5221 CGGCAAAATC CCTTATAAAT CAAAAGAATA GACCGAGATA GGGTTGAGTG TTGTTCCAGT

     5281 TTGGAACAAG AGTCCACTAT TAAAGAACGT GGACTCCAAC GTCAAAGGGC GAAAAACCGT

     5341 CTATCAGGGC GATGGCCCAC TACGTGAACC ATCACCCTAA TCAAGTTTTT TGGGGTCGAG

     5401 GTGCCGTAAA GCACTAAATC GGAACCCTAA AGGGAGCCCC CGATTTAGAG CTTGACGGGG

     5461 AAAGCCGGCG AACGTGGCGA GAAAGGAAGG GAAGAAAGCG AAAGGAGCGG GCGCTAGGGC

     5521 GCTGGCAAGT GTAGCGGTCA CGCTGCGCGT AACCACCACA CCCGCCGCGC TTAATGCGCC

     5581 GCTACAGGGC GCGTCCCATT CGCCA

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pET-48b(+)其他大肠杆菌表达载体：