pSpCas9n(BB)-2A-Puro (PX462) V2.0
(Plasmid #62987)

• Purpose: Cas9n (D10A nickase mutant) from S. pyogenes with 2A-Puro, and cloning backbone for sgRNA (V2.0)
• Depositing Lab: Feng Zhang
• Publication: Ran et al Nat Protoc. 2013 Nov;8(11):2281-308. doi: 10.1038/nprot.2013.143. Epub 2013 Oct 24.

Backbone

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

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: hSpCas9n-2A-Puro
• Alt name: SpCas9n-2A-Puro V2.0
• Alt name: Cas9n
• Alt name: PX462 V2.0
• Species: Synthetic
• Insert Size (bp): 6000
• Mutation: Cas9 D10A nickase mutant
• Tags / Fusion Proteins:
  • 3XFLAG (N terminal on insert)
  • 2A-Puro (C terminal on insert)

Resource Information

• Supplemental Documents:
  • Zhang lab general target sequence cloning protocol
• Addgene Notes:
  • Addgene diagnostic digest
• Articles Citing this Plasmid:
  • 88 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

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

The guide sequence is cloned into this plasmid using BbsI sites. Please see the cloning protocol provided by the Zhang lab for more details.