pRAN357
(Plasmid #120898)

• Purpose: E. coli-C. difficile shuttle vector: tetracycline-inducible promoter (Ptet) driving expression of cyan fluorescent protein (CfpOpt), multiple-cloning site for fusion to N-terminus of target protein
• Depositing Labs: Craig Ellermeier, David S. Weiss
• Publication: Ransom et al J Bacteriol. 2014 Jun;196(12):2290-300. doi: 10.1128/JB.00038-14. Epub 2014 Apr 11.

Backbone

• Vector backbone: pRFP185
• Backbone size w/o insert (bp): 6400
• Total vector size (bp): 7959
• Modifications to backbone: Ptet::cfpOpt-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: cyan fluorescent protein, codon-optimized for low GC organisms, multiple-cloning site for protein fusion
• Alt name: Ptet::cfpOpt-mcs
• Species: Synthetic
• 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:
  • pRAN357.gb

Terms and Licenses

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