pJK220
(Plasmid #71123)

• Purpose: Expression of E. coli pantotheinate kinase (H6PanK, R175H mutant) in E. coli
• Depositing Lab: T. J. Kappock
• Publication: Francois et al Biochemistry. 2006 Nov 14;45(45):13487-99.

Backbone

• Vector backbone: pET28a
• Backbone size w/o insert (bp): 5369
• Total vector size (bp): 6296
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: pantothenate kinase
• Alt name: CoaA
• Alt name: panK
• Species: Escherichia coli
• Mutation: Arg175 is mutated to a His
• GenBank ID: 62288041
• Promoter: T7
• Tag / Fusion Protein:
  • H6 (N terminal on backbone)

Cloning Information

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

Terms and Licenses

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

Depositor Comments

PCR product was first ligated into pDrive plasmid and found to lack desired 3' EcoRI site. Subcloned into pET23a using the pDrive-encoded EcoRI site. Therefore an additional 18 bp is present in the 3'-UTR between the stop codon and EcoRI site.