pRAN473
(Plasmid #120813)

• Purpose: E. coli-C. difficile shuttle vector: tetracycline-inducible promoter (Ptet) driving expression of red fluorescent protein (mCherryOpt), multiple cloning site for fusion to N-terminus of target protein
• Depositing Labs: Craig Ellermeier, David S. Weiss
• Publication: Ransom et al Appl Environ Microbiol. 2015 Mar;81(5):1652-60. doi: 10.1128/AEM.03446-14. Epub 2014 Dec 19.

Backbone

• Vector backbone: pRFP185
• Backbone size w/o insert (bp): 6400
• Total vector size (bp): 7950
• Modifications to backbone: Ptet::mCherryOpt-mcs
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 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: red fluorescent protein (mCherry), codon-optimized for C. difficile, multiple-cloning site for protein fusion
• Alt name: mCherryOpt-mcs
• Species: Synthetic
• Insert Size (bp): 1583
• Promoter: Ptet

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SacI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: unknown
• 3′ sequencing primer: unknown

Resource Information

• Supplemental Documents:
  • pRAN473.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry