pJK556
(Plasmid #74277)

• Purpose: Produces formylglycinamide ribonucleotide amidotransferase, N-terminal His6 tag (H6EcPurL)
• Depositing Lab: T. J. Kappock
• Publication: Purine biosynthesis (unpublished)

Backbone

• Vector backbone: pET22b
• Total vector size (bp): 9306
• Modifications to backbone: pET22b-HT adds a 17-residue sequence including a N-terminal His6 tag
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: formylglycinamidine ribonucleotide synthetase
• Alt name: large PurL
• Species: Escherichia coli
• Mutation: changes alanine-1246 to serine, numerous silent mutations
• GenBank ID: WP_000970138.1
• Promoter: T7
• Tag / Fusion Protein:
  • His6 (N terminal on backbone)

Cloning Information

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

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Based on a plasmid obtained from Jamie Williamson (Scripps).

Terms and Licenses

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

Depositor Comments

Repaired form of a plasmid described originally in Schultheisz HL, Szymczyna BR, Scott LG, Williamson JR (2008) Pathway engineered enzymatic de novo purine nucleotide synthesis. ACS Chem Biol. 3, 499-511. doi: 10.1021/cb800066p