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pET-42b(+)

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At a Glance

pET-42b(+), Image provided by Novagen

Type

Cloning, Expression

Origin of Replication

f1
pBR322

Copy #

~40

Markers

Kanamycin

Link to Sequence

Novagen

Features

T7 Promoter
N Terminal GST Tag
S Tag
MCS
lacI
C Terminal His Tag

Accession Number




Notes

The pET-42b(+) has the popular GST fusion tag for enhanced production and solubility.

The pET-42 series is designed for cloning and high-level expression of peptide sequences fused with the 220 aa GST•Tag™ protein. The T7 expression region is reversed on the vector map due to the sequencing numbered by the pBR322 convention. 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, T7 terminator primer should be used for single-stranded sequencing. When cloning into the PshaI site, the vector encoded sequence can be removed completely followed by the cleaving of the GST fusion protein with Factor Xa.

History

Sources

Novagen

References

[1]

  1. Wei, S et al. (2005) Reactive site mutations in tissue inhibitor of metalloproteinase-3 disrupt inhibition of matrix metalloproteinases but not tumor necrosis factor-alpha-converting enzyme. J. Biol. Chem. 280 32877-82 PubMed EcoliWiki page

Sequence

       1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 
      61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt 
     121 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt 
     181 gcggccgcaa gcttgtcgac ggagctcgcc tgcaggcgcg ccaaggcctg tacagaattc 
     241 ggatccccga tatccatgga cccgcgtccc tcaataccgg agccaccacc ggtacccaga 
     301 tctgggctgt ccatgtgctg gcgttcgaat ttagcagcag cggtttcttt cataccaatt 
     361 gcagtactac cgcgtggcac cagacccgcg gagtgatggt gatggtgatg accagaacca 
     421 ctagttgaac catccgattt tggaggatgg tcgccaccac caaacgtggc ttgccagccc 
     481 tgcaaaggcc atgctatata cttgctggat ttcaagtact tatcaatttg tgggatagct 
     541 tcaatacgtt ttttaaaaca aactaatttt gggaacgcat ccaggcacat tgggtccatg 
     601 tataaaacaa catcaagagc gtcatacaac atgaagtcag gatgggttac atgatcacca 
     661 tttaaatatg ttttatgaca taaacgatct tcgaacattt tcagcatttc aggtagcttg 
     721 ctaagaaaat caactttgag agtttcaaag tctttactat atgcaattct cgaaacaccg 
     781 tatctaatat ccaaaaccgc tccttcaagc attgaaatct ctgcacgctc ttttggacaa 
     841 ccacccaaca tgttgtgctt gtcagctata taacgtatga tggccataga ctgtgttaat 
     901 ttaacatcac catcaatata ataaggaaga ttgggaaact ccaaacccaa ttcaaacttt 
     961 ttgtttcgcc atttatcacc ttcatcgcgc tcatacaaat gctcttcata tttttcttca 
    1021 agatattcca aaagaagtcg agtgggttgc acaaggccct taattttcca ataacctagt 
    1081 ataggggaca tatgtatatc tccttcttaa agttaaacaa aattatttct agaggggaat 
    1141 tgttatccgc tcacaattcc cctatagtga gtcgtattaa tttcgcggga tcgagatcga 
    1201 tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg 
    1261 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca 
    1321 tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg 
    1381 ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac 
    1441 tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgagatccc ggacaccatc 
    1501 gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg 
    1561 gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat 
    1621 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa 
    1681 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg 
    1741 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg 
    1801 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg 
    1861 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa 
    1921 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa 
    1981 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac 
    2041 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg 
    2101 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt 
    2161 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa 
    2221 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc 
    2281 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt 
    2341 accgagtccg ggctgcgcgt tggtgcggac atctcggtag tgggatacga cgataccgaa 
    2401 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg 
    2461 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag 
    2521 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc 
    2581 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa 
    2641 agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg caccgggatc 
    2701 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat 
    2761 gactagcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac 
    2821 tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac 
    2881 gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga 
    2941 aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg 
    3001 gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat 
    3061 ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa 
    3121 ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat 
    3181 cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa 
    3241 aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact 
    3301 caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga 
    3361 tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 
    3421 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 
    3481 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag 
    3541 cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg 
    3601 caccatatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc 
    3661 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 
    3721 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 
    3781 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 
    3841 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 
    3901 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 
    3961 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 
    4021 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 
    4081 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 
    4141 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 
    4201 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 
    4261 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 
    4321 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 
    4381 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 
    4441 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 
    4501 tggtcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc 
    4561 catattcaac gggaaacgtc ttgctctagg ccgcgattaa attccaacat ggatgctgat 
    4621 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga 
    4681 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc 
    4741 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg 
    4801 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc 
    4861 gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat 
    4921 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac 
    4981 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat 
    5041 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg 
    5101 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat 
    5161 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc 
    5221 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca 
    5281 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag 
    5341 tttcatttga tgctcgatga gtttttctaa gaattaattc atgagcggat acatatttga 
    5401 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 
    5461 tgaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 
    5521 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 
    5581 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 
    5641 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 
    5701 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 
    5761 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 
    5821 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 
    5881 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgcca