pJK633
(Plasmid #72397)

• Purpose: Produces Escherichia coli aminoimidazole ribonucleotide synthetase, N-terminal His6 tag (H6EcPurM)
• Depositing Lab: T. J. Kappock
• Publication: Purine biosynthesis (unpublished)

Backbone

• Vector backbone: pET22b
• Total vector size (bp): 6490
• 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: phosphoribosylaminoimidazole synthetase
• Alt name: purM
• Alt name: phosphoribosylformylglycinamidine cycloligase, JW2484
• Alt name: aminoimidazole ribonucleotide synthetase
• Species: Escherichia coli str. K-12 substr. MG1655
• GenBank ID: NP_416994.1 AAC75552
• Promoter: T7
• Tag / Fusion Protein:
  • His6 (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: EcoRI (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: Heather L. Schultheisz (PI:James R. Williamson)

Terms and Licenses

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

Depositor Comments

Source plasmid contained a Gly324>Asp mutation that we fixed using QuikChange.

Reference for source plasmid:
Schultheisz, H. L., Szymczyna, B. R., Scott, L. G., and Williamson, J. R. (2008) Pathway engineered enzymatic de novo purine nucleotide synthesis ACS Chem. Biol. 3 499 511 [DOI: 10.1021/cb800066p]