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pSpCas9n(BB)-2A-Puro (PX462)
(Plasmid #48141)

Full plasmid sequence is not available for this item.

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 48141 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    PX462
  • Total vector size (bp) 9200
  • Vector type
    Mammalian Expression, CRISPR
  • Selectable markers
    Puromycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    Stbl3
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    hSpCas9n-2A-Puro
  • Alt name
    SpCas9n-2A-Puro
  • Alt name
    Cas9n
  • Alt name
    PX462
  • Species
    Synthetic
  • Insert Size (bp)
    6000
  • Mutation
    Cas9 D10A nickase mutant
  • Promoter Cbh
  • Tags / Fusion Proteins
    • 3XFLAG (N terminal on insert)
    • 2A-Puro (C terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site AgeI (not destroyed)
  • 3′ cloning site EcoRI (not destroyed)
  • 5′ sequencing primer TTTATGGCGAGGCGGCGG
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Note that this is the plasmid used in the original publication. The Puro gene in this plasmid has a point mutation that renders it less effective in some cell lines. The new version of the plasmid has corrected that point mutation, thus improving Puro selection.

For more information, please see the following web page: http://crispr.genome-engineering.org

For more information on Zhang Lab CRISPR Plasmids please refer to: http://www.addgene.org/crispr/zhang/

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:

    pSpCas9n(BB)-2A-Puro (PX462) was a gift from Feng Zhang (Addgene plasmid # 48141)
  • For your References section:

    Genome engineering using the CRISPR-Cas9 system. Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F. Nat Protoc. 2013 Nov;8(11):2281-308. doi: 10.1038/nprot.2013.143. Epub 2013 Oct 24. 10.1038/nprot.2013.143 PubMed 24157548