pET15b-17X-PTDH
(Plasmid #166786)

• Purpose: Phosphite dehydrogenase for NAD(P)H regeneration
• Depositing Lab: Wilfred van der Donk
• Publication: Yang et al J Am Chem Soc. 2015 Oct 7;137(39):12426-9. doi: 10.1021/jacs.5b05207. Epub 2015 Sep 24.

Backbone

• Vector backbone: pET15b
• Backbone manufacturer: Addgene
• Backbone size w/o insert (bp): 5708
• Total vector size (bp): 6711
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: Phosphite dehydroenase
• Alt name: PTDH
• Species: Pseudomonas stutzeri
• Insert Size (bp): 1008
• Mutation: A196R, T201S, A328T, E352N, and C356D- please see depositor comments
• GenBank ID: WP_106442756
• Promoter: T7
• Tag / Fusion Protein:
  • His6 (N terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: T7 terminator

Resource Information

• Supplemental Documents:
  • pET15b-17X-PTDH.gb

Terms and Licenses

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

Depositor Comments

The only mutation of interest is A196R which allows the enzyme to use both NADH and NADPH as a cofactor for catalysis. Dep. confirms the additional mutations T201S, A328T, E352N, and C356D do not affect plasmid function. The enzyme is very active.