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pET-44a(+)

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At a Glance

pET-44a(+), Image provided by Novagen

Type

Cloning and Expression

Origin of Replication

f1
pBR322

Copy #

~40

Markers

Ampicillin

Link to Sequence

Novagen

Features

T7 Promoter
N and C Terminal His Tag
S Tag
Nus Tag
MCS
C Terminal HSV Tag
lacI

Accession Number




Notes

The pET-44a(+) vector is specially designed for cloning and high-level expression of polypeptide sequences fused with the 495 aa NusA (Nus•Tag) protein. When E.coli cytoplasm is expressed as NusA fusions, the solubility of the various target protein in E.Coli cytoplasm is expressed. pET-43.1(+) vector series derived the pET-44(+) series which encodes an additional N-Terminal His•Tag sequence. When cloning into the PshA I or Sma I sites, the vector encoded sequence can be completely removed by the cleavage of the Nus•Tag fusion protein with enterokinase or thrombin, respectively.

History

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

Sources

Novagen

References

[1] [2]

  1. Hong, KU et al. (2007) Functional importance of the anaphase-promoting complex-Cdh1-mediated degradation of TMAP/CKAP2 in regulation of spindle function and cytokinesis. Mol. Cell. Biol. 27 3667-81 PubMed EcoliWiki page
  2. Agetsuma, M et al. (2005) The ubiquitin-proteasome pathway is involved in rapid degradation of phosphoenolpyruvate carboxylase kinase for C4 photosynthesis. Plant Cell Physiol. 46 389-98 PubMed EcoliWiki page

Sequence

       1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 
      61 ggggttatgc tagttattgc tcagcggata catatttgaa tgtatttaga aaaataaaca 
     121 aaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat 
     181 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt 
     241 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga 
     301 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc 
     361 cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag 
     421 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc 
     481 agcctaggaa cgcccaactt aattaacatt agtggtggtg gtggtggtgc tcgagatcct 
     541 ctgggtcttc aggagcgagt tctggctggc ttgcacgtgt atacagctgt gcggccgcaa 
     601 gcttcgaacg cgtatcgatg gtaccgtcga cgtcctgcag gcgcgcctgt acagaattcg 
     661 gatccacgag ctcccggact cttgtcgtcg tcatcaatcg taccagaacc cgcgctgccc 
     721 cgggggacca gaccagttgg cggtggcgag tccatgtgct ggcgttcaaa tttcgcagca 
     781 gcggtttctt tacccgcgga gtgatggtga tggtgatgac cagaaccact agtcgcttcg 
     841 tcaccgaacc agcaaatatt acgggcagcc ataatcagtg ctccggcttt ttcgtcggtc 
     901 aacccttcga tatcagccag atcatcaatg ccctgttcgg cgagatcttc cagcgtacaa 
     961 acgccacggg cggccagttt gaatgccaaa tcacgatcta ccccttcaag gttcagcaga 
    1021 tcgtcagccg gtttgttatc accgaggctt tcttcctggg cctgtgcaat ggtggccagt 
    1081 gcatttttag cacgctcgcg cagtgcttca acggtcggct catcaaggcc ttcgatttcc 
    1141 aacagctctt tcatcggcac ataggccaat tcttccagcg tcgagaagcc ttcttctacc 
    1201 agaacagtcg cgaagtcttc gtcgatgtcg agatatttgg tgaaggtgtc gatcgctgcg 
    1261 tgcgcttccg cctgatgctt agcttgcagg tcgtcaacgg tcatcacgtt gagttcccaa 
    1321 ccgctcagtt gcgaagccag acgcacgttc tgaccgttac ggccaatcgc ctgcgccaga 
    1381 ttaccggctt caacggcgat gtccatggtg tgtttatctt catccaccac gatagaagca 
    1441 acgtctgccg gtgccattgc gttaatcacg aactgcgccg ggttatcatc ccacaggacg 
    1501 atatcgatac gctcgccacc cagttcagta gacaccgcct gaacacgcgc gccacgcata 
    1561 cctacgcaag cacctaccgg atcgatacgt ttatcgttgg ttttcaccgc gattttcgca 
    1621 cgagaacccg gatcgcgagc cgctgcttta atttcaatca cttcttcgcc gatttctggc 
    1681 acttcaatac ggaacagttc gatcagcatt tccggcttgg aacgagtgac gaacagttgc 
    1741 gcgccacgcg cttccgggcg aacggaatag agcacgccac gaacgcggtc gccagggcgg 
    1801 aagttttcac gcggcagcat atcttcgcgc aggatcacgg cttcagcgtt gttgcccaga 
    1861 tccagagaga tgttgtcgcg gtttactttt ttcaccacgc cggtgatgat ttcaccttcg 
    1921 tgttcacgga actgatcaac caccatcgca cgttcggctt cacgcacttt ctgcacgata 
    1981 acctgttttg ccgtctgggt agtgatacgg tcaaaggtaa cagactcaat ctgatcttca 
    2041 acgtaatcgc ccaggttcag gctttcatct tcataacgtg cggcttcaag ggtgatttcc 
    2101 ttggtcggct gggtgacttc atcaacaact aaccagcgac ggaaagtgtc aaaatcaccg 
    2161 cttttgcgat cgatctgtac gcggacgtcg atctcttgtt catatttttt ctttgttgct 
    2221 gtcgccagcg cgctttccaa tgcttcgaaa atcttctcgc gaggtagcgc cttttcattg 
    2281 gatacggctt caactacagc caaaatttct ttgttcatac tagaatggtg atggtggtga 
    2341 tggctgctgc ccatatgtat atctccttct taaagttaaa caaaattatt tctagagggg 
    2401 aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat 
    2461 cgatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg 
    2521 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc 
    2581 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg 
    2641 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac 
    2701 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc 
    2761 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc 
    2821 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct 
    2881 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa 
    2941 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg 
    3001 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg 
    3061 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg 
    3121 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg 
    3181 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg 
    3241 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc 
    3301 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca 
    3361 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg 
    3421 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg 
    3481 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc 
    3541 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc 
    3601 attaccgagt ccgggctgcg cgttggtgcg gacatctcgg tagtgggata cgacgatacc 
    3661 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg 
    3721 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat 
    3781 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc 
    3841 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg 
    3901 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg 
    3961 atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg 
    4021 catgactagc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct 
    4081 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa 
    4141 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct 
    4201 ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg 
    4261 ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt 
    4321 gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag 
    4381 taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg 
    4441 tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga 
    4501 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa 
    4561 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc 
    4621 tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca 
    4681 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 
    4741 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg 
    4801 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga 
    4861 gtgcaccata tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca 
    4921 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 
    4981 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 
    5041 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 
    5101 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 
    5161 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 
    5221 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 
    5281 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 
    5341 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 
    5401 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 
    5461 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 
    5521 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 
    5581 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 
    5641 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 
    5701 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 
    5761 ttttggtcat gacctatttg tttatttttc taaatacatt caaatatgta tccgctcatg 
    5821 agacaataac cctgataaat gcttcaatca tgattgaaaa aggaagagta tgagtattca 
    5881 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca 
    5941 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta 
    6001 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt 
    6061 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc 
    6121 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc 
    6181 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc 
    6241 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa 
    6301 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga 
    6361 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat 
    6421 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca 
    6481 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc 
    6541 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat 
    6601 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag 
    6661 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa 
    6721 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca 
    6781 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatgc catcgtgccg 
    6841 ccagaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc 
    6901 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc 
    6961 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac 
    7021 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca 
    7081 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg 
    7141 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag 
    7201 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc 
    7261 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc a