MSP1101
(Plasmid #65773)

• Purpose: Human expression vector for SpCas9 VRER variant: CMV-T7-humanSpCas9(D1135V/G1218R/R1335E/T1337R)-NLS-3xFLAG (VRER variant)
• Depositing Lab: Keith Joung
• Publication: Kleinstiver et al Nature. 2015 Jun 22. doi: 10.1038/nature14592.

Backbone

• Vector backbone: JDS246
• Backbone manufacturer: Joung lab (Addgene Plasmid # 43861)
• Vector type: Mammalian Expression, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: mammalian codon-optimized Streptococcus pyogenes Cas9 (D1135V/G1218R/R1335E/T1337R)-NLS-3XFlag
• Mutation: D1135V, G1218R, R1335E, and T1337R mutations in Cas9
• Promoter: CMV & T7
• Tags / Fusion Proteins:
  • NLS (C terminal on insert)
  • 3x FLAG (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: BGH-rev

Resource Information

• Supplemental Documents:
  • Annotated Genbank file
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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