pJB06
(Plasmid #190480)

• Purpose: Base Cas9 expression plasmid for 2-plasmid C. difficile mutagenesis system
• Depositing Lab: Joseph Sorg
• Publication: Brehm et al Microbiol Resour Announc. 2022 Dec 15;11(12):e0083322. doi: 10.1128/mra.00833-22. Epub 2022 Nov 7.

Backbone

• Vector backbone: pHN149
• Backbone size w/o insert (bp): 7084
• Total vector size (bp): 12489
• Vector type: E. coli / B. subtilis - C. difficile shuttle vector

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: Cas9
• Species: S. pyogenes
• Insert Size (bp): 4107
• Promoter: xylR-driven promoter

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: TCAAGGAGGGTAAAGAGGAGAGttaattaa
• 3′ sequencing primer: tgccaagcttgcatgtctgcaggcctcgag

Gene/Insert 2

• Gene/Insert name: XylR
• Insert Size (bp): 1227

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: caatttttttatcaggaaacagctatgacc
• 3′ sequencing primer: TAATAATAAAATATTTAATTATCTTTGTCA

Resource Information

• Supplemental Documents:
  • pjb06(1).gb

Terms and Licenses

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