pRPF144
(Plasmid #106372)

• Purpose: C. difficile constitutive expression system. Shuttle plasmid containing constitutive Pcwp2 -gusA
• Depositing Labs: Robert Fagan, Neil Fairweather
• Publication: Fagan et al J Biol Chem. 2011 Aug 5;286(31):27483-93. doi: 10.1074/jbc.M111.263889. Epub 2011 Jun 9.

Backbone

• Vector backbone: pMTL960
• Backbone size w/o insert (bp): 6218
• Total vector size (bp): 8258
• Vector type: E. coli - C. difficile shuttle vector

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 15 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Low copy in C. difficile following conjugation and selected with thiamphenicol
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Pcwp2-gusA
• Insert Size (bp): 2040
• Mutation: codon optimised for C. difficile
• Entrez Gene: gusA (a.k.a. ECO111_2086)
• Promoter: Pcwp2

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: KpnI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: CACCTCCTTTTTGACTTTAAGCCTACGAATACC
• 3′ sequencing primer: CACCGACGAGCAAGGCAAGACCG

Resource Information

• Supplemental Documents:
  • C. difficile expression plasmids
  • pRPF144 corrected.gb

Terms and Licenses

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