PurposeCRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 5F5.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||73432||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Backbone manufacturerLuciano Marraffini (Addgene plasmid # 46569)
- Backbone size w/o insert (bp) 9296
Vector typeBacterial Expression, CRISPR, Synthetic Biology
Growth in Bacteria
Copy numberLow Copy
Gene/Insert nameRepressor 5F5 (orthogonal T7-lac repressor)
- Promoter Constitutive wild-type S. pyogenes promoter
- Cloning method Restriction Enzyme
- 5′ cloning site BsaI (not destroyed)
- 3′ cloning site BsaI (not destroyed)
- 5′ sequencing primer dCas9-F3 (CACGCATTGATTTGAGTCAGC) (Common Sequencing Primers)
Terms and Licenses
Also encodes dCas9 and tracrRNA.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pdCas9-M-5F5 was a gift from Mattheos Koffas (Addgene plasmid # 73432 ; http://n2t.net/addgene:73432 ; RRID:Addgene_73432)
For your References section:Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Cress BF, Jones JA, Kim DC, Leitz QD, Englaender JA, Collins SM, Linhardt RJ, Koffas MA. Nucleic Acids Res. 2016 Apr 13. pii: gkw231. 10.1093/nar/gkw231 PubMed 27079979