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pET-41b(+)

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At a Glance

pET-41b(+), Image provided by Novagen

Type

Cloning, Expression

Origin of Replication

f1
pBR322

Copy #

~40

Markers

Kanamycin

Link to Sequence

Novagen

Features

T7 promoter [1]
C terminal His Tag
N Terminal GST Tag
MCS
lacI
S Tag

Accession Number




Notes

pET-41b(+) has the popular GST fusion tag for enhanced production and solubility.

The pET-41 series is designed for cloning and high-level expression of peptide sequences fused with the 220 aa GST•Tag protein. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is included. 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 Enterokinase.

History

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Sources

Novagen

References

[2] [3]

  1. Torres, VJ et al. (2005) Functional properties of the p33 and p55 domains of the Helicobacter pylori vacuolating cytotoxin. J. Biol. Chem. 280 21107-14 PubMed EcoliWiki page
  2. Adams, H et al. (2006) Interaction of TonB with the outer membrane receptor FpvA of Pseudomonas aeruginosa. J. Bacteriol. 188 5752-61 PubMed EcoliWiki page
  3. Schmelz, EA et al. (2006) Fragments of ATP synthase mediate plant perception of insect attack. Proc. Natl. Acad. Sci. U.S.A. 103 8894-9 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 tatccatggg actcttgtcg tcgtcatcac cggagccacc accggtaccc 
     301 agatctgggc tgtccatgtg ctggcgttcg aatttagcag cagcggtttc tttcatacca 
     361 attgcagtac taccgcgtgg caccagaccc gcggagtgat ggtgatggtg atgaccagaa 
     421 ccactagttg aaccatccga ttttggagga tggtcgccac caccaaacgt ggcttgccag 
     481 ccctgcaaag gccatgctat atacttgctg gatttcaagt acttatcaat ttgtgggata 
     541 gcttcaatac gttttttaaa acaaactaat tttgggaacg catccaggca cattgggtcc 
     601 atgtataaaa caacatcaag agcgtcatac aacatgaagt caggatgggt tacatgatca 
     661 ccatttaaat atgttttatg acataaacga tcttcgaaca ttttcagcat ttcaggtagc 
     721 ttgctaagaa aatcaacttt gagagtttca aagtctttac tatatgcaat tctcgaaaca 
     781 ccgtatctaa tatccaaaac cgctccttca agcattgaaa tctctgcacg ctcttttgga 
     841 caaccaccca acatgttgtg cttgtcagct atataacgta tgatggccat agactgtgtt 
     901 aatttaacat caccatcaat ataataagga agattgggaa actccaaacc caattcaaac 
     961 tttttgtttc gccatttatc accttcatcg cgctcataca aatgctcttc atatttttct 
    1021 tcaagatatt ccaaaagaag tcgagtgggt tgcacaaggc ccttaatttt ccaataacct 
    1081 agtatagggg acatatgtat atctccttct taaagttaaa caaaattatt tctagagggg 
    1141 aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat 
    1201 cgatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg 
    1261 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc 
    1321 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg 
    1381 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac 
    1441 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc 
    1501 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc 
    1561 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct 
    1621 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa 
    1681 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg 
    1741 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg 
    1801 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg 
    1861 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg 
    1921 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg 
    1981 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc 
    2041 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca 
    2101 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg 
    2161 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg 
    2221 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc 
    2281 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc 
    2341 attaccgagt ccgggctgcg cgttggtgcg gacatctcgg tagtgggata cgacgatacc 
    2401 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg 
    2461 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat 
    2521 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc 
    2581 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg 
    2641 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg 
    2701 atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg 
    2761 catgactagc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct 
    2821 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa 
    2881 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct 
    2941 ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg 
    3001 ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt 
    3061 gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag 
    3121 taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg 
    3181 tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga 
    3241 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa 
    3301 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc 
    3361 tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca 
    3421 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 
    3481 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg 
    3541 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga 
    3601 gtgcaccata tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca 
    3661 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 
    3721 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 
    3781 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 
    3841 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 
    3901 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 
    3961 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 
    4021 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 
    4081 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 
    4141 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 
    4201 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 
    4261 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 
    4321 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 
    4381 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 
    4441 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 
    4501 ttttggtcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg 
    4561 agccatattc aacgggaaac gtcttgctct aggccgcgat taaattccaa catggatgct 
    4621 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat 
    4681 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt 
    4741 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt 
    4801 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc 
    4861 cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt 
    4921 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt 
    4981 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt 
    5041 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa 
    5101 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt 
    5161 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga 
    5221 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct 
    5281 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg 
    5341 cagtttcatt tgatgctcga tgagtttttc taagaattaa ttcatgagcg gatacatatt 
    5401 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 
    5461 acctgaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag 
    5521 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac 
    5581 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga 
    5641 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc 
    5701 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg 
    5761 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa 
    5821 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac 
    5881 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc ca