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pET-42a(+)

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At a Glance

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pET-42a(+), Image provided by Novagen

Type

Cloning, Expression

Origin of Replication

f1
pBR322

Copy #

~40

Markers

Kanamycin

Link to Sequence

Novagen

Features

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

Accession Number




Notes

The pET-42a(+) has the popular GST fusion tag for enhanced production and solubility.

The pET-42 series is designed for cloning and high-level expression of peptide sequences fused with the 220 aa GST•Tag™ protein. 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 Factor Xa.

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. Kulman, JD et al. (2007) Proline-rich Gla protein 2 is a cell-surface vitamin K-dependent protein that binds to the transcriptional coactivator Yes-associated protein. Proc. Natl. Acad. Sci. U.S.A. 104 8767-72 PubMed EcoliWiki page
  2. Beernink, PT et al. (2005) Specificity of protein interactions mediated by BRCT domains of the XRCC1 DNA repair protein. J. Biol. Chem. 280 30206-13 PubMed EcoliWiki page
  3. Chellappan, P et al. (2005) MicroRNA-binding viral protein interferes with Arabidopsis development. Proc. Natl. Acad. Sci. U.S.A. 102 10381-6 PubMed EcoliWiki page
  4. Fotso, P et al. (2005) Cog1p plays a central role in the organization of the yeast conserved oligomeric Golgi complex. J. Biol. Chem. 280 27613-23 PubMed EcoliWiki page
  5. Li, L et al. (2005) Clarification of cinnamoyl co-enzyme A reductase catalysis in monolignol biosynthesis of Aspen. Plant Cell Physiol. 46 1073-82 PubMed EcoliWiki page
  6. Liedtke, CM et al. (2005) Role for protein phosphatase 2A in the regulation of Calu-3 epithelial Na+-K+-2Cl-, type 1 co-transport function. J. Biol. Chem. 280 25491-8 PubMed EcoliWiki page
  7. Madsen, KL et al. (2005) Molecular determinants for the complex binding specificity of the PDZ domain in PICK1. J. Biol. Chem. 280 20539-48 PubMed EcoliWiki page
  8. Srivastava, SK et al. (2005) NAD+-dependent DNA Ligase (Rv3014c) from Mycobacterium tuberculosis. Crystal structure of the adenylation domain and identification of novel inhibitors. J. Biol. Chem. 280 30273-81 PubMed EcoliWiki page
  9. Yang, HY et al. (2005) Kinesin-1 mediates translocation of the meiotic spindle to the oocyte cortex through KCA-1, a novel cargo adapter. J. Cell Biol. 169 447-57 PubMed EcoliWiki page
  10. Zong, X et al. (2005) A novel mechanism of modulation of hyperpolarization-activated cyclic nucleotide-gated channels by Src kinase. J. Biol. Chem. 280 34224-32 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 tatcccatgg acccgcgtcc ctcaataccg gagccaccac cggtacccag 
     301 atctgggctg tccatgtgct ggcgttcgaa tttagcagca gcggtttctt tcataccaat 
     361 tgcagtacta ccgcgtggca ccagacccgc ggagtgatgg tgatggtgat gaccagaacc 
     421 actagttgaa ccatccgatt ttggaggatg gtcgccacca ccaaacgtgg cttgccagcc 
     481 ctgcaaaggc catgctatat acttgctgga tttcaagtac ttatcaattt gtgggatagc 
     541 ttcaatacgt tttttaaaac aaactaattt tgggaacgca tccaggcaca ttgggtccat 
     601 gtataaaaca acatcaagag cgtcatacaa catgaagtca ggatgggtta catgatcacc 
     661 atttaaatat gttttatgac ataaacgatc ttcgaacatt ttcagcattt caggtagctt 
     721 gctaagaaaa tcaactttga gagtttcaaa gtctttacta tatgcaattc tcgaaacacc 
     781 gtatctaata tccaaaaccg ctccttcaag cattgaaatc tctgcacgct cttttggaca 
     841 accacccaac atgttgtgct tgtcagctat ataacgtatg atggccatag actgtgttaa 
     901 tttaacatca ccatcaatat aataaggaag attgggaaac tccaaaccca attcaaactt 
     961 tttgtttcgc catttatcac cttcatcgcg ctcatacaaa tgctcttcat atttttcttc 
    1021 aagatattcc aaaagaagtc gagtgggttg cacaaggccc ttaattttcc aataacctag 
    1081 tataggggac atatgtatat ctccttctta aagttaaaca aaattatttc tagaggggaa 
    1141 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatcg 
    1201 atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt 
    1261 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc 
    1321 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc 
    1381 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta 
    1441 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat 
    1501 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag 
    1561 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta 
    1621 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa 
    1681 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc 
    1741 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc 
    1801 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc 
    1861 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca 
    1921 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga 
    1981 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa 
    2041 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt 
    2101 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg 
    2161 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga 
    2221 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat 
    2281 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat 
    2341 taccgagtcc gggctgcgcg ttggtgcgga catctcggta gtgggatacg acgataccga 
    2401 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg 
    2461 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca 
    2521 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc 
    2581 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga 
    2641 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat 
    2701 ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca 
    2761 tgactagcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta 
    2821 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa 
    2881 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg 
    2941 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct 
    3001 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga 
    3061 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta 
    3121 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta 
    3181 tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa 
    3241 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac 
    3301 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg 
    3361 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 
    3421 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 
    3481 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta 
    3541 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt 
    3601 gcaccatata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg 
    3661 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 
    3721 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 
    3781 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 
    3841 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 
    3901 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 
    3961 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 
    4021 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 
    4081 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 
    4141 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 
    4201 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 
    4261 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 
    4321 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 
    4381 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 
    4441 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 
    4501 ttggtcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag 
    4561 ccatattcaa cgggaaacgt cttgctctag gccgcgatta aattccaaca tggatgctga 
    4621 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg 
    4681 attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc 
    4741 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc 
    4801 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc 
    4861 cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga 
    4921 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa 
    4981 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga 
    5041 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat 
    5101 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga 
    5161 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat 
    5221 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc 
    5281 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca 
    5341 gtttcatttg atgctcgatg agtttttcta agaattaatt catgagcgga tacatatttg 
    5401 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 
    5461 ctgaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct 
    5521 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg 
    5581 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact 
    5641 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac 
    5701 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga 
    5761 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga 
    5821 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca 
    5881 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca