p3R112C/T180CBM3
(Plasmid #85090)

• Purpose: Expresses PCNA3 variant (R112C/T180C) fused to P450 BM3 variant (A74G/C773S/C810S) 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): 5333
• Total vector size (bp): 9269
• 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 P450 BM3
• Alt name: PCNA1(G108C)-P450 BM3
• Species: Synthetic; Sulfolobus solfataricus P2, Bacillus megaterium
• Insert Size (bp): 3936
• Mutation: R112C/T180C in PCNA3, A74G/C773S/C810S in P450BM3
• 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 plasmid was obtained from p3R112C/T180C-GG (Plasmid #85093) and pBsaBM3 (Plasmid #85102) by Golden Gate assembly.