pIA123
(Plasmid #234038)

• Purpose: E. coli-C. difficile shuttle vector for CRISPR editing in C. difficile. Pxyl::Cas9-opt Pgdh::sgRNA-cdr_0985. PstI site to add DNA for homology directed repair. MscI and NotI for replacement of sgRNA.
• Depositing Lab: Craig Ellermeier
• Publication: Bollinger et al Proc Natl Acad Sci U S A. 2024 Aug 20;121(34):e2408540121. doi: 10.1073/pnas.2408540121. Epub 2024 Aug 16.

Backbone

• Vector backbone: pIA33
• Backbone size w/o insert (bp): 6300
• Total vector size (bp): 12940
• Vector type: Bacterial Expression, CRISPR

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: Cas9
• Alt name: Cas9 nuclease, codon optimized for C. difficile
• Species: Streptococcus pyogenes
• Promoter: Pxyl

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: unknown
• 3′ sequencing primer: unknown

Gene/Insert 2

• Gene/Insert name: sgRNA
• Alt name: single guide RNA, targeting cdr_0985
• Species: Streptococcus pyogenes
• Promoter: Pgdh

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: unknown
• 3′ sequencing primer: unknown

Resource Information

• Supplemental Documents:
  • pIA123.gb

Terms and Licenses

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