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pET-28a(+)

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At a Glance

pET-28a-c(+), image provided by Novagen

Type

Cloning, Expression

Origin of Replication

f1
pBR322

Copy #

~40

Markers

Kanamycin

Link to Sequence

Novagen

Features

T7 Promoter
His Tag
MCS
lacI

Accession Number




Notes

The pET-28a(+) vector carries an N-terminal His•Tag/thrombin/T7•Tag configuration in addition to an optional C-terminal His•Tag sequence. Unique sites are shown on the circle map. The T7 expression region is reversed on the circular map due to the sequence numbering from 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, the T7 terminator primer should be used when performing 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] [11] [12] [13]

  1. Jin, Y et al. (2002) Identification of hNopp140 as a binding partner for doxorubicin with a phage display cloning method. Chem. Biol. 9 157-62 PubMed EcoliWiki page
  2. Ludlam, WH et al. (2002) The acetyltransferase activity of CBP is required for wingless activation and H4 acetylation in Drosophila melanogaster. Mol. Cell. Biol. 22 3832-41 PubMed EcoliWiki page
  3. Renshaw, PS et al. (2002) Conclusive evidence that the major T-cell antigens of the Mycobacterium tuberculosis complex ESAT-6 and CFP-10 form a tight, 1:1 complex and characterization of the structural properties of ESAT-6, CFP-10, and the ESAT-6*CFP-10 complex. Implications for pathogenesis and virulence. J. Biol. Chem. 277 21598-603 PubMed EcoliWiki page
  4. Seybert, A et al. (2002) Biochemical characterisation of the clamp/clamp loader proteins from the euryarchaeon Archaeoglobus fulgidus. Nucleic Acids Res. 30 4329-38 PubMed EcoliWiki page
  5. Okubo, T et al. (2001) Down-regulation of promoter 1.3 activity of the human aromatase gene in breast tissue by zinc-finger protein, snail (SnaH). Cancer Res. 61 1338-46 PubMed EcoliWiki page
  6. Kim, JW et al. (2000) Human RNA lariat debranching enzyme cDNA complements the phenotypes of Saccharomyces cerevisiae dbr1 and Schizosaccharomyces pombe dbr1 mutants. Nucleic Acids Res. 28 3666-73 PubMed EcoliWiki page
  7. 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
  8. Nakielny, S et al. (1999) Nup153 is an M9-containing mobile nucleoporin with a novel Ran-binding domain. EMBO J. 18 1982-95 PubMed EcoliWiki page
  9. Orphanides, G et al. (1999) The chromatin-specific transcription elongation factor FACT comprises human SPT16 and SSRP1 proteins. Nature 400 284-8 PubMed EcoliWiki page
  10. Kenny, B et al. (1997) Enteropathogenic E. coli (EPEC) transfers its receptor for intimate adherence into mammalian cells. Cell 91 511-20 PubMed EcoliWiki page
  11. Peña, SV et al. (1997) Processing, subcellular localization, and function of 519 (granulysin), a human late T cell activation molecule with homology to small, lytic, granule proteins. J. Immunol. 158 2680-8 PubMed EcoliWiki page
  12. Gudi, R et al. (1995) Diversity of the pyruvate dehydrogenase kinase gene family in humans. J. Biol. Chem. 270 28989-94 PubMed EcoliWiki page
  13. Xu, Y et al. (1995) High-yield bacterial expression, purification, and functional reconstitution of the tricarboxylate transport protein from rat liver mitochondria. Biochem. Biophys. Res. Commun. 207 783-9 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 tccgcgaccc atttgctgtc caccagtcat gctagccata 
     241 tggctgccgc gcggcaccag gccgctgctg tgatgatgat gatgatggct gctgcccatg 
     301 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca 
     361 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc 
     421 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc 
     481 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc 
     541 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca 
     601 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg 
     661 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt 
     721 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc 
     781 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt 
     841 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc 
     901 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct 
     961 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat 
    1021 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg 
    1081 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat 
    1141 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt 
    1201 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca 
    1261 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc 
    1321 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa 
    1381 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat 
    1441 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct 
    1501 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg 
    1561 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat 
    1621 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg 
    1681 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact 
    1741 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc 
    1801 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca 
    1861 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga 
    1921 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac 
    1981 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca 
    2041 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat 
    2101 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg 
    2161 gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc 
    2221 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac 
    2281 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa 
    2341 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct 
    2401 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta 
    2461 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca 
    2521 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag 
    2581 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa 
    2641 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc 
    2701 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg 
    2761 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc 
    2821 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 
    2881 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 
    2941 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg 
    3001 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatat gcggtgtgaa 
    3061 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc 
    3121 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 
    3181 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 
    3241 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 
    3301 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 
    3361 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 
    3421 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 
    3481 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 
    3541 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 
    3601 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 
    3661 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 
    3721 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 
    3781 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 
    3841 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 
    3901 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgaa caataaaact 
    3961 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc 
    4021 ttgctctagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc 
    4081 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc 
    4141 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat 
    4201 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg 
    4261 tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt 
    4321 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg 
    4381 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct 
    4441 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga 
    4501 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc 
    4561 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg 
    4621 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct 
    4681 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca 
    4741 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga 
    4801 gtttttctaa gaattaattc atgagcggat acatatttga atgtatttag aaaaataaac 
    4861 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaaattgta aacgttaata 
    4921 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg 
    4981 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc 
    5041 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa 
    5101 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt 
    5161 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac 
    5221 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta 
    5281 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg 
    5341 cgccgctaca gggcgcgtcc cattcgcca