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pET-24a(+)

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At a Glance

pET-24a(+) Image made with PlasMapper

Type

Expression

Origin of Replication

f1, pBR322

Copy #

~40

Markers

Kanamycin

Link to Sequence

Novagen

Features

T7 promoter
Multiple cloning sites
His•Tag coding sequence
T7 terminator
lacI coding sequence

Accession Number




Notes

The pET-24a-d(+) vectors possess an N-terminal sequence that codes for a T7 Tag. It also comes with an optional C-terminal His Tag sequence. The only difference between each individual plasmid from a-d is the selectable marker. In pET-24a(+) the selectable marker is kanamycin. On the vector ap the T7 repression region is opposite a normal region meaning it is reversed. This is a characteristic of the BR322 numbering convention. When exposed to helper phage, the fl origin produces virions containing single-dtranded DNA. These single stranded DNA correspond to the coding strand. The virions are a product of the orientation of the f1 origin.

History

All pET-24a-d(+) plasmids are the same except for their selectable markers. (kanamycin vs. ampicillin)

Sources

Novagen

References

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

  1. Sukumar, N et al. (2007) Differential Bvg phase-dependent regulation and combinatorial role in pathogenesis of two Bordetella paralogs, BipA and BcfA. J. Bacteriol. 189 3695-704 PubMed EcoliWiki page
  2. Boanca, G et al. (2006) Uncoupling the enzymatic and autoprocessing activities of Helicobacter pylori gamma-glutamyltranspeptidase. J. Biol. Chem. 281 19029-37 PubMed EcoliWiki page
  3. Gakh, O et al. (2006) Mitochondrial iron detoxification is a primary function of frataxin that limits oxidative damage and preserves cell longevity. Hum. Mol. Genet. 15 467-79 PubMed EcoliWiki page
  4. Sosnovtsev, SV et al. (2006) Cleavage map and proteolytic processing of the murine norovirus nonstructural polyprotein in infected cells. J. Virol. 80 7816-31 PubMed EcoliWiki page
  5. Beharry, Z & Palzkill, T (2005) Functional analysis of active site residues of the fosfomycin resistance enzyme FosA from Pseudomonas aeruginosa. J. Biol. Chem. 280 17786-91 PubMed EcoliWiki page
  6. Islam, MN et al. (2005) Construction of new forms of pyruvate carboxylase to assess the allosteric regulation by acetyl-CoA. Protein Eng. Des. Sel. 18 71-8 PubMed EcoliWiki page
  7. Nakamura, Y et al. (2005) A novel phosphatidylcholine-hydrolyzing phospholipase C induced by phosphate starvation in Arabidopsis. J. Biol. Chem. 280 7469-76 PubMed EcoliWiki page
  8. Rider, SD Jr et al. (2005) The protozoan parasite Cryptosporidium parvum possesses two functionally and evolutionarily divergent replication protein A large subunits. J. Biol. Chem. 280 31460-9 PubMed EcoliWiki page
  9. Skretas, G & Wood, DW (2005) Regulation of protein activity with small-molecule-controlled inteins. Protein Sci. 14 523-32 PubMed EcoliWiki page
  10. Cooper, KW & Baneyx, F (2001) Escherichia coli FtsH (HflB) degrades a membrane-associated TolAI-II-beta-lactamase fusion protein under highly denaturing conditions. Protein Expr. Purif. 21 323-32 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 tgtatatctc cttcttaaag ttaaacaaaa ttatttctag aggggaattg ttatccgctc 
     301 acaattcccc tatagtgagt cgtattaatt tcgcgggatc gagatctcga tcctctacgc 
     361 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc 
     421 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg 
     481 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc 
     541 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat 
     601 gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct 
     661 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac 
     721 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg 
     781 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg 
     841 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc 
     901 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga 
     961 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg 
    1021 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga 
    1081 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg 
    1141 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc 
    1201 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg 
    1261 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata 
    1321 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca 
    1381 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc 
    1441 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc 
    1501 gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata 
    1561 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc 
    1621 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac 
    1681 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg 
    1741 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc 
    1801 aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg 
    1861 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca 
    1921 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc 
    1981 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta 
    2041 ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc 
    2101 ggcgagaagc aggccattat cgccggcatg gcggccccac gggtgcgcat gatcgtgctc 
    2161 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca 
    2221 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca 
    2281 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc 
    2341 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct 
    2401 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc 
    2461 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca 
    2521 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga 
    2581 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc 
    2641 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat 
    2701 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc 
    2761 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc 
    2821 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt 
    2881 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact 
    2941 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatata tgcggtgtga 
    3001 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct 
    3061 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 
    3121 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 
    3181 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 
    3241 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 
    3301 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 
    3361 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 
    3421 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 
    3481 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 
    3541 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 
    3601 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 
    3661 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 
    3721 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 
    3781 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 
    3841 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga acaataaaac 
    3901 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt 
    3961 cttgctctag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg 
    4021 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg 
    4081 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga 
    4141 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc 
    4201 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg 
    4261 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc 
    4321 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc 
    4381 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg 
    4441 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct 
    4501 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg 
    4561 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc 
    4621 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc 
    4681 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg 
    4741 agtttttcta agaattaatt catgagcgga tacatatttg aatgtattta gaaaaataaa 
    4801 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt aaacgttaat 
    4861 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc 
    4921 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt 
    4981 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa 
    5041 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg 
    5101 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga 
    5161 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct 
    5221 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat 
    5281 gcgccgctac agggcgcgtc ccattcgcca