pCas9cur
(Plasmid #71538)

• Purpose: Constitutive expression of Cas9 gene and inducible expression of the plasmid curing system for CRISPR mediated genome editing of E. coli.
• Depositing Lab: Tao Chen
• Publication: Li et al Metab Eng. 2015 Sep;31:13-21. doi: 10.1016/j.ymben.2015.06.006. Epub 2015 Jun 30.

Backbone

• Vector backbone: pSC101ts
• Vector type: Bacterial Expression, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert 1

• Gene/Insert name: Cas9
• Species: S. pyogenes

Gene/Insert 2

• Gene/Insert name: Plasmid curing system
• Alt name: gRNA targeting bla gene
• Promoter: pBAD

Cloning Information for Gene/Insert 2

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

Resource Information

• Supplemental Documents:
  • pCas9cur genbank
• A portion of this plasmid was derived from a plasmid made by: Cas9 is from pCas9, Addgene plasmid 42876.

Terms and Licenses

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

Depositor Comments

The plasmid curing system consists of an arabinose inducible promoter, PBAD to express a gRNA targeting the bla gene on the gRNA plasmid. When induced, the bla-targeting gRNA is expressed, leading Cas9 to cleave the gRNA plasmid and resulting in plasmid elimination.