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pET PI-SceI C-His K301A
(Plasmid #195611)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 195611 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pET-23a
  • Backbone manufacturer
    Novagen
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    PI-SceI
  • Alt name
    VDE
  • Species
    S. cerevisiae (budding yeast)
  • Mutation
    A822G, A823C, A824T
  • Promoter T7

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site Unknown (unknown if destroyed)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Expresses the excised intein of the VMA1 gene product including amino-terminal methionine-cysteine residues. Contains silent AvrII and RsrII sites near the borders of the N-terminal LAGLIDADG motif and silent EaeI and SnaBI sites near the borders of the C-terminal motif. Also contains silent mutations that introduce ApaI, BssHII, BstEII, BsiWI, MluI and SpeI sites. Expresses PI-SceI containing a carboxyl-terminal histidine tag and a K301A mutation in the PI-SceI ORF.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pET PI-SceI C-His K301A was a gift from Frederick Gimble (Addgene plasmid # 195611 ; http://n2t.net/addgene:195611 ; RRID:Addgene_195611)

  • For your References section:

    Amino acid residues in both the protein splicing and endonuclease domains of the PI-SceI intein mediate DNA binding. He Z, Crist M, Yen H, Duan X, Quiocho FA, Gimble FS. J Biol Chem. 1998 Feb 20;273(8):4607-15. doi: 10.1074/jbc.273.8.4607. 10.1074/jbc.273.8.4607 PubMed 9468518
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