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pET-41a(+)

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At a Glance

pET-41a(+), 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
C Terminal His Tag
S Tag
lacI
MCS

Accession Number




Notes

pET-41a(+) has the popular GST fusion tag for enhanced production and solubility.

The pET-41 series is designed for cloning and high-level expression of peptide sequences fused with the 220 aa GST•Tag protein. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is included. 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 Enterokinase.

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 gactcttgtc gtcgtcatca ccggagccac caccggtacc 
     301 cagatctggg ctgtccatgt gctggcgttc gaatttagca gcagcggttt ctttcatacc 
     361 aattgcagta ctaccgcgtg gcaccagacc cgcggagtga tggtgatggt gatgaccaga 
     421 accactagtt gaaccatccg attttggagg atggtcgcca ccaccaaacg tggcttgcca 
     481 gccctgcaaa ggccatgcta tatacttgct ggatttcaag tacttatcaa tttgtgggat 
     541 agcttcaata cgttttttaa aacaaactaa ttttgggaac gcatccaggc acattgggtc 
     601 catgtataaa acaacatcaa gagcgtcata caacatgaag tcaggatggg ttacatgatc 
     661 accatttaaa tatgttttat gacataaacg atcttcgaac attttcagca tttcaggtag 
     721 cttgctaaga aaatcaactt tgagagtttc aaagtcttta ctatatgcaa ttctcgaaac 
     781 accgtatcta atatccaaaa ccgctccttc aagcattgaa atctctgcac gctcttttgg 
     841 acaaccaccc aacatgttgt gcttgtcagc tatataacgt atgatggcca tagactgtgt 
     901 taatttaaca tcaccatcaa tataataagg aagattggga aactccaaac ccaattcaaa 
     961 ctttttgttt cgccatttat caccttcatc gcgctcatac aaatgctctt catatttttc 
    1021 ttcaagatat tccaaaagaa gtcgagtggg ttgcacaagg cccttaattt tccaataacc 
    1081 tagtataggg gacatatgta tatctccttc ttaaagttaa acaaaattat ttctagaggg 
    1141 gaattgttat ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga 
    1201 tcgatctcga tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg 
    1261 gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg 
    1321 ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg 
    1381 ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta 
    1441 ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaga tcccggacac 
    1501 catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt 
    1561 cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc 
    1621 ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga 
    1681 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact 
    1741 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc 
    1801 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt 
    1861 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc 
    1921 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt 
    1981 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat 
    2041 caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc 
    2101 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct 
    2161 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg 
    2221 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg 
    2281 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc 
    2341 cattaccgag tccgggctgc gcgttggtgc ggacatctcg gtagtgggat acgacgatac 
    2401 cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct 
    2461 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa 
    2521 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac 
    2581 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact 
    2641 ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcaccgg 
    2701 gatctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg 
    2761 gcatgactag catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc 
    2821 ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca 
    2881 aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc 
    2941 tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct 
    3001 gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag 
    3061 tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca 
    3121 gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg 
    3181 gtatcattac ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg 
    3241 aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga 
    3301 aactcaacga gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg 
    3361 ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac 
    3421 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag 
    3481 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac 
    3541 gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag 
    3601 agtgcaccat atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc 
    3661 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 
    3721 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 
    3781 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 
    3841 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 
    3901 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 
    3961 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 
    4021 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc 
    4081 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 
    4141 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 
    4201 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 
    4261 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag 
    4321 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 
    4381 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 
    4441 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 
    4501 attttggtca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat 
    4561 gagccatatt caacgggaaa cgtcttgctc taggccgcga ttaaattcca acatggatgc 
    4621 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta 
    4681 tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt 
    4741 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct 
    4801 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat 
    4861 ccccgggaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt 
    4921 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt 
    4981 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt 
    5041 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga 
    5101 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact 
    5161 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg 
    5221 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc 
    5281 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt 
    5341 gcagtttcat ttgatgctcg atgagttttt ctaagaatta attcatgagc ggatacatat 
    5401 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 
    5461 cacctgaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca 
    5521 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga 
    5581 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg 
    5641 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat 
    5701 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag 
    5761 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga 
    5821 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa 
    5881 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg cca