pC3R112C/T180C
(Plasmid #85088)

• Purpose: Expresses PCNA3 variant (R112C/T180C) fused to P450cam (CYP101A1) in E. coli
• Depositing Lab: Teruyuki Nagamune
• Publication: Tan et al Angew Chem Int Ed Engl. 2016 Oct 26. doi: 10.1002/anie.201608033.

Backbone

• Vector backbone: pET-28b(+)
• Backbone manufacturer: Merck Millipore
• Backbone size w/o insert (bp): 5332
• Total vector size (bp): 7366
• Modifications to backbone: N/A
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Supplement with 1% glucose is recommended to suppress leaky expression.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: The R112C/T180C variant of PCNA3 fused to P450cam
• Alt name: P450cam-PCNA3(R112C/T180C)
• Species: Synthetic; Sulfolobus solfataricus P2, Pseudomonas putida
• Insert Size (bp): 2034
• Mutation: R112C/T180C in PCNA3
• Promoter: T7 promoter
• Tag / Fusion Protein:
  • His6 tag (N terminal on backbone)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7 Terminal

Terms and Licenses

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

Depositor Comments

The plamid was obtained from pGS-3R112C/T180C (Plasmid #85095) and pBsaP450camGG (Plasmid #85101) by Golden Gate assembly.