pSpCas9(BB)-2A-Puro (PX459)
(Plasmid #48139)

• Purpose: NOTE: A new version of this plasmid is now available. See Addgene plasmid 62988.
• 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: PX459
• 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: hSpCas9-2A-Puro
• Alt name: SpCas9-2A-Puro
• Alt name: Cas9
• Alt name: PX459
• Species: Synthetic
• Insert Size (bp): 6000
• 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

Resource Information

• Supplemental Documents:
  • PX459 pSpCas9(BB)-T2A-Puro annotated Genbank file
• Addgene Notes:
  • Addgene Diagnostic Digest
• Articles Citing this Plasmid:
  • 545 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/.