pDC10
(Plasmid #75077)

• Purpose: Produces Escherichia coli glycinamide ribonucleotide synthetase (H6EcPurD)
• Depositing Labs: T. J. Kappock, JoAnne Stubbe
• Publication: Purine biosynthesis (unpublished)

Backbone

• Vector backbone: pET28a
• Backbone size w/o insert (bp): 5369
• Total vector size (bp): 6910
• Modifications to backbone: N-terminal His6 tag sequence contains a Ser11>Asn mutation. NdeI site replaced with EcoRI sequence which replaces Met1 codon with Glu-Phe codons.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): JM109
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: phosphoribosylglycinamide synthetase
• Alt name: purD
• Alt name: JW3969
• Alt name: b4005
• Species: Escherichia coli
• Mutation: adds unique SacII site at Thr95-Ala96 by a silent mutation in Ala96 codon
• GenBank ID: NP_418433
• Entrez Gene: purD (a.k.a. b4005, ECK3997, JW3969, adth)
• Promoter: T7
• Tag / Fusion Protein:
  • His6 (N terminal on backbone)

Cloning Information

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

Terms and Licenses

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

Depositor Comments

Constructed by Danny Crawford and T. J. Kappock in the laboratory of JoAnne Stubbe (MIT).

In JM109 strain.