pEF_dCas9
(Plasmid #68416)

• Purpose: Transient expression of Sp-dCas9 in mammalian cells, under an EF1-alpha promoter.
• Depositing Lab: John Rinn
• Publication: Shechner et al Nat Methods. 2015 Jul;12(7):664-70. doi: 10.1038/nmeth.3433. Epub 2015 Jun 1.

Backbone

• Vector backbone: pNEB193
• Backbone manufacturer: NEB
• 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: dCas9
• Mutation: D10A/H841A
• Promoter: human EF1[alpha]
• Tag / Fusion Protein:
  • 3xFLAG (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Unknown (unknown if destroyed)
• 3′ cloning site: Unknown (unknown if destroyed)
• 5′ sequencing primer: EF-1a-F
• 3′ sequencing primer: BGH-rev

Resource Information

• Supplemental Documents:
  • pEF_dCas9 genbank
• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

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

Depositor Comments

Contains NLS sequences at the N- and C-terminus of the dCas9 protein. The C-terminal NLS immediately preceedes the 3xFLAG tag