AW01_pU.CAG.eSpCas9(1.1).rBGpA
(Plasmid #199255)

• Purpose: Expression construct encoding the high specificity eSpCas9(1.1) nuclease
• Depositing Lab: Manuel Goncalves
• Publication: Chen et al Nucleic Acids Res. 2016 Jun 8. pii: gkw524.

Backbone

• Vector backbone: pBR322
• Backbone size w/o insert (bp): 4965
• Total vector size (bp): 9360
• Vector type: Mammalian Expression, CRISPR

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: High specificity eSpCas9(1.1) nuclease
• Species: Streptococcus pyogenes
• Insert Size (bp): 4266
• Mutation: K848A K1003A R1060A
• Promoter: CAG promoter
• Tags / Fusion Proteins:
  • 3XFLAG epitope (N terminal on insert)
  • SV40 NLS (N terminal on insert)
  • Nucleoplasmin NLS (C terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: M13 rev
• 3′ sequencing primer: M13 fwd

Resource Information

• Supplemental Documents:
  • AW01_pU.CAG.eSpCas9(1.1).rBGpA.gbk
• A portion of this plasmid was derived from a plasmid made by: The SpCas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system, mutated to K848A, K1003A and R1060A to improve targeting specificity (Slaymaker et al., Science 2016 351:84-88) was inserted into a pCAG.rBGpA expression plasmid backbone.

Terms and Licenses

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