pJK597
(Plasmid #71121)

• Purpose: Produces E coli formyl-CoA:oxalate CoA-transferase (H6YfdW) in E. coli
• Depositing Lab: T. J. Kappock
• Publication: Mullins et al Protein Sci. 2012 May;21(5):686-96. doi: 10.1002/pro.2054. Epub 2012 Mar 29.

Backbone

• Vector backbone: pET28a
• Total vector size (bp): 5943
• Modifications to backbone: During cloning 0.66 kb from the vector untranslated region (a region that includes the T7 terminator, F1 origin, and M13 origin) was deleted by an unknown mechanism.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: formyl-CoA:oxalate CoA-transferase
• Alt name: YfdW
• Alt name: frc
• Alt name: formyl-CoA transferase, NAD(P)-binding
• Species: Escherichia coli str. K-12 substr. MG1655
• GenBank ID: NP_416875
• Promoter: T7
• Tag / Fusion Protein:
  • His6 (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: EcoRI (destroyed during cloning)
• 5′ sequencing primer: T7 promoter

Terms and Licenses

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

Depositor Comments

An unexplained deletion event removed the F1 ori, M13 ori region, the T7 terminator, and C-terminal His-tag region. Instead we see a sequence of 29 nt that does not correspond to any obvious source. The net plasmid is 629 nt shorter than expected. The major practical consequence of this is there is no terminator -- of any kind -- oriented parallel to the expressed gene. The expressed protein ORF appears to be completely correct.