pdCas9-M-3A2
(Plasmid #73435)

• Purpose: CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 3A2.
• Depositing Lab: Mattheos Koffas
• Publication: Cress et al Nucleic Acids Res. 2016 Apr 13. pii: gkw231.

Backbone

• Vector backbone: pdCas9
• Backbone manufacturer: Luciano Marraffini (Addgene plasmid # 46569)
• Backbone size w/o insert (bp): 9296
• Vector type: Bacterial Expression, CRISPR, Synthetic Biology

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: Repressor 3A2 (orthogonal T7-lac repressor)
• gRNA/shRNA sequence: ATTAATACGACTCACTATCA
• Species: Synthetic
• Promoter: Constitutive wild-type S. pyogenes promoter

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsaI (not destroyed)
• 3′ cloning site: BsaI (not destroyed)
• 5′ sequencing primer: dCas9-F3 (CACGCATTGATTTGAGTCAGC)

Terms and Licenses

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

Depositor Comments

Also encodes dCas9 and tracrRNA.