pDNR-SpCas9-Gem
(Plasmid #80424)

• Purpose: Used for in vitro transcription of SpCas9-Gem mRNA. Cas9 is fused to Geminin peptide and is degraded during G0/G1 phases of the cell-cycle to minimize indels caused by non-homologous end joining
• Depositing Labs: Sara Howden, Melissa Little
• Publication: Howden et al Stem Cell Reports. 2016 Sep 13;7(3):508-17. doi: 10.1016/j.stemcr.2016.07.001. Epub 2016 Aug 4.

Backbone

• Vector backbone: pDNR-Dual
• Backbone manufacturer: Clontech
• Vector type: CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: May get better pDNA yield if grown at 30 degrees
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: SpCas9-Gem
• Species: S.pyogenes
• Insert Size (bp): 4479
• Mutation: codon optimized for human
• Promoter: T7
• Tag / Fusion Protein:
  • Geminin (C terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: taatacgactcactatag

Resource Information

• Supplemental Documents:
  • pDNR-SpCas9Gem
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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