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Addgene

pET15b I-SceI (Ctermhis) K122I
(Plasmid #195614)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 195614 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pET-15b
  • 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
    I-SceI
  • Species
    S. cerevisiae (budding yeast)
  • Mutation
    C2458G, A2826T, A2827T
  • Promoter T7

Cloning Information

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

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Originally derived from pSCM525, a plasmid from Dr. Bernard Dujon at the Institute Pasteur.

Terms and Licenses

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

Depositor Comments

Expresses the I-SceI homing endonuclease under the control of the phage T7 promoter. The sequence is codon optimized for expression in E. coli and is derived from plasmid pSCM525 (Dr. Bernard Dujon, Institut Pasteur). The protein contains a carboxyl-terminal histidine tag for purification. The expressed protein contains the K122I mutation that results in DNA nicking activity. An appended methionine-histidine sequence is present at the amino terminus.

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:

    pET15b I-SceI (Ctermhis) K122I was a gift from Frederick Gimble (Addgene plasmid # 195614 ; http://n2t.net/addgene:195614 ; RRID:Addgene_195614)

  • For your References section:

    Engineering variants of the I-SceI homing endonuclease with strand-specific and site-specific DNA-nicking activity. Niu Y, Tenney K, Li H, Gimble FS. J Mol Biol. 2008 Sep 26;382(1):188-202. doi: 10.1016/j.jmb.2008.07.010. Epub 2008 Jul 11. 10.1016/j.jmb.2008.07.010 PubMed 18644379
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