pRPF215
(Plasmid #106377)

• Purpose: C. difficile transposon mutagenesis system. Shuttle vector containing tet-inducible Himar1 transposase and custom ErmR transposon
• Depositing Labs: Robert Fagan, Neil Fairweather
• Publication: Dembek et al MBio. 2015 Feb 24;6(2):e02383. doi: 10.1128/mBio.02383-14.

Backbone

• Vector backbone: pMTL960
• Backbone size w/o insert (bp): 6191
• Total vector size (bp): 9231
• 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: Ptet-himar1-Ter(slpA)-transposon
• Insert Size (bp): 3040
• Mutation: Codon optimised for C. difficile
• Promoter: Ptet

Cloning Information

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

Resource Information

• Supplemental Documents:
  • pRPF215 corrected.gb
  • Use of pRPF215
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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