p2T-CAG-SpCas9-BlastR
(Plasmid #107190)

• Purpose: Confers constitutive expression of SpCas9
• Depositing Lab: Richard Sherwood
• Publication: Shen et al Nature. 2018 Nov;563(7733):646-651. doi: 10.1038/s41586-018-0686-x. Epub 2018 Nov 7.

Backbone

• Vector backbone: sgBbsI (p2Tol-U6-2xBbsI-sgRNA-HygR)
• Backbone manufacturer: Richard Sherwood Lab
• Backbone size w/o insert (bp): 6982
• Total vector size (bp): 12083
• Modifications to backbone: The sequence between Tol2 sites was replaced with a CAGGS promoter, Cas9 sequence, and Blasticidin resistance cassette.
• Vector type: Mammalian Expression, Bacterial Expression, CRISPR
• Selectable markers: Blasticidin

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: SpCas9
• Insert Size (bp): 4101
• Entrez Gene: NEWENTRY
• Promoter: Chicken ß-actin
• Tag / Fusion Protein:
  • 3xFLAG

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: MfeI (not destroyed)
• 3′ cloning site: AgeI (not destroyed)
• 5′ sequencing primer: CGAGGTCGACGGTATCG
• 3′ sequencing primer: CAAGTTAACAACAACAATTGCATTCATTTT

Resource Information

• Supplemental Documents:
  • p2T_CAG_Cas9MA_BlastR.gb
• Addgene Notes:
  • Addgene Diagnostic Digest
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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