pCAS9counter
(Plasmid #107192)

• Purpose: Expresses CAS9 and sgRNA for targeted counterselection in S. aureus
• Depositing Lab: Steve Salipante
• Publication: Penewit et al MBio. 2018 Feb 20;9(1). pii: mBio.00067-18. doi: 10.1128/mBio.00067-18.

Backbone

• Vector backbone: pCN-50
• Backbone size w/o insert (bp): 5750
• Total vector size (bp): 7788
• Modifications to backbone: Addition of EF2132 and tetR. Replacement of cat194 gene with ermC gene from pCN33. Replacement of pT181cop-634ts with E194ts replicon.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Erythromycin selection in S. aureus, Ampicillin selection in E. coli. Temperature sensitive replicon must be maintained at 32 degrees in S. aureus. Stable at 37 degrees in E. coli.
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: Cas9
• Species: S pyogenes
• Insert Size (bp): 4297
• Promoter: prab17

Cloning Information for Gene/Insert 1

• Cloning method: Unknown
• 5′ sequencing primer: None
• 3′ sequencing primer: None

Gene/Insert 2

• Gene/Insert name: sgRNA
• Insert Size (bp): 324
• Promoter: prab17

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: None
• 3′ sequencing primer: None

Resource Information

• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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