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pET-29b(+)

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At a Glance

pET-29a-c(+), image provided by Novagen

Type

Cloning, Expression

Origin of Replication

f1
pBR322

Copy #

~40

Markers

Kanamycin

Link to Sequence

NCBI

Features

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

Accession Number

15897468



Notes

The pET-29b(+) vector carries an N-terminal S•Tag™/thrombin configuration including an optional C-terminal His•Tag® sequence. The T7 expression region is reversed on the vector map due to the sequencing numbered by the pBR322 convention. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is included. 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.

History

You can help EcoliWiki by writing about the history of this plasmid.

Sources

Novagen

References

[1] [2] [3] [4] [5] [6] [7] [8] [9] [10]

  1. Fischbach, MA et al. (2007) Directed evolution can rapidly improve the activity of chimeric assembly-line enzymes. Proc. Natl. Acad. Sci. U.S.A. 104 11951-6 PubMed EcoliWiki page
  2. Hsu, IaW et al. (2005) Phosphorylation of Y14 modulates its interaction with proteins involved in mRNA metabolism and influences its methylation. J. Biol. Chem. 280 34507-12 PubMed EcoliWiki page
  3. Jilaveanu, LB et al. (2005) Dimeric SecA is essential for protein translocation. Proc. Natl. Acad. Sci. U.S.A. 102 7511-6 PubMed EcoliWiki page
  4. Korepanova, A et al. (2005) Cloning and expression of multiple integral membrane proteins from Mycobacterium tuberculosis in Escherichia coli. Protein Sci. 14 148-58 PubMed EcoliWiki page
  5. Müllegger, J et al. (2005) Engineering of a thioglycoligase: randomized mutagenesis of the acid-base residue leads to the identification of improved catalysts. Protein Eng. Des. Sel. 18 33-40 PubMed EcoliWiki page
  6. Nam, JO et al. (2005) Regulation of tumor angiogenesis by fastatin, the fourth FAS1 domain of betaig-h3, via alphavbeta3 integrin. Cancer Res. 65 4153-61 PubMed EcoliWiki page
  7. Zhang, W et al. (2005) Biophysical and kinetic characterization of HemAT, an aerotaxis receptor from Bacillus subtilis. Biophys. J. 88 2801-14 PubMed EcoliWiki page
  8. Zito, CR et al. (2005) Role of a conserved glutamate residue in the Escherichia coli SecA ATPase mechanism. J. Biol. Chem. 280 14611-9 PubMed EcoliWiki page
  9. Kanaya, E et al. (1999) Characterization of the transcriptional activator CBF1 from Arabidopsis thaliana. Evidence for cold denaturation in regions outside of the DNA binding domain. J. Biol. Chem. 274 16068-76 PubMed EcoliWiki page
  10. Huang, B et al. () NMR structure and mutagenesis of the Fas (APO-1/CD95) death domain. Nature 384 638-41 PubMed EcoliWiki page

Sequence

       1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 
      61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt 
     121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt 
     181 cgacggagct cgaattcgga tccgatatcg ccatggaacc gcgtggcacc agggtaccca 
     241 gatctgggct gtccatgtgc tggcgttcga atttagcagc agcggtttct ttcatatgta 
     301 tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa 
     361 ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tcgatctcga tcctctacgc 
     421 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc 
     481 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg 
     541 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc 
     601 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat 
     661 gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct 
     721 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac 
     781 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg 
     841 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg 
     901 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc 
     961 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga 
    1021 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg 
    1081 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga 
    1141 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg 
    1201 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc 
    1261 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg 
    1321 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata 
    1381 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca 
    1441 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc 
    1501 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc 
    1561 gcgttggtgc ggacatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata 
    1621 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc 
    1681 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac 
    1741 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg 
    1801 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc 
    1861 aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg 
    1921 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca 
    1981 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc 
    2041 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta 
    2101 ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc 
    2161 ggcgagaagc aggccattat cgccggcatg gcggccccac gggtgcgcat gatcgtgctc 
    2221 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca 
    2281 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca 
    2341 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc 
    2401 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct 
    2461 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc 
    2521 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca 
    2581 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga 
    2641 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc 
    2701 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat 
    2761 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc 
    2821 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc 
    2881 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt 
    2941 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact 
    3001 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatata tgcggtgtga 
    3061 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct 
    3121 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 
    3181 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 
    3241 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 
    3301 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 
    3361 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 
    3421 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 
    3481 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 
    3541 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 
    3601 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 
    3661 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 
    3721 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 
    3781 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 
    3841 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 
    3901 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga acaataaaac 
    3961 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt 
    4021 cttgctctag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg 
    4081 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg 
    4141 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga 
    4201 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc 
    4261 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg 
    4321 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc 
    4381 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc 
    4441 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg 
    4501 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct 
    4561 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg 
    4621 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc 
    4681 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc 
    4741 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg 
    4801 agtttttcta agaattaatt catgagcgga tacatatttg aatgtattta gaaaaataaa 
    4861 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt aaacgttaat 
    4921 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc 
    4981 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt 
    5041 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa 
    5101 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg 
    5161 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga 
    5221 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct 
    5281 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat 
    5341 gcgccgctac agggcgcgtc ccattcgcca