pAF302
(Plasmid #137752)

• Purpose: Intracellular control plasmid for HiBiTopt experiments
• Depositing Lab: Wiep Klaas Smits
• Publication: Oliveira Paiva et al J Bacteriol. 2020 Aug 31. pii: JB.00771-19. doi: 10.1128/JB.00771-19.

Backbone

• Vector backbone: pRPF185
• Backbone manufacturer: Fagan and Fairweather
• Backbone size w/o insert (bp): 7200
• Total vector size (bp): 7566
• Vector type: Bacterial Expression
• Selectable markers: thiamphenicol

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: hupA-linker-hibitopt
• Species: Clostridioides difficile
• Insert Size (bp): 366
• GenBank ID: YP_001090017.1
• Promoter: Ptet
• Tag / Fusion Protein:
  • HiBiTopt (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SacI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: CACCTCCTTTTTGACTTTAAGCCTACGAATACC (NF793)
• 3′ sequencing primer: CACCGACGAGCAAGGCAAGACCG (NF794)

Resource Information

• Supplemental Documents:
  • pAF302.gb

Terms and Licenses

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