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Addgene

Cas9-NAT
(Plasmid #64329)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 64329 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pRS414-TEF1p-Cas9-CYC1t
  • Backbone manufacturer
    George Church lab (Addgene Plasmid #43802)
  • Backbone size w/o insert (bp) 9232
  • Total vector size (bp) 10446
  • Vector type
    Yeast Expression, CRISPR
  • Selectable markers
    Nourseothricin(clonNat)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Top10 also works
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    Human-optimized S. pyogenes Cas9 with Clonnat marker gene expression cassette
  • Insert Size (bp)
    1206

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site SnaBI (not destroyed)
  • 3′ cloning site MfeI (not destroyed)
  • 5′ sequencing primer tctagagcggccgcTACGTAgatctgtttagcttgccttg
  • 3′ sequencing primer tctagagcggccgcCAATTGcgttttcgacactggatggc
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Plasmid p414-TEF1p-Cas9-CYC1t from George Church lab purchased from Addgene
  • Articles Citing this Plasmid

Terms and Licenses

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

    Cas9-NAT was a gift from Yong-Su Jin (Addgene plasmid # 64329 ; http://n2t.net/addgene:64329 ; RRID:Addgene_64329)
  • For your References section:

    Construction of a quadruple auxotrophic mutant of an industrial polyploid saccharomyces cerevisiae strain by using RNA-guided Cas9 nuclease. Zhang GC, Kong II, Kim H, Liu JJ, Cate JH, Jin YS. Appl Environ Microbiol. 2014 Dec;80(24):7694-701. doi: 10.1128/AEM.02310-14. Epub 2014 Oct 3. 10.1128/AEM.02310-14 PubMed 25281382