pMT643
(Plasmid #8610)

• Depositing Lab: Robert Hartley
• Publication: Hartley et al Biochemistry 1993 Jun 15;32(23):5978-84.

Backbone

• Vector backbone: pUC19
• Backbone size w/o insert (bp): 2641
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: barstar A
• Species: Bacillus amyloliquefaciens
• Insert Size (bp): 439
• Mutation: Two cysteines of barstar changed to alanine, C40A and C82A
• GenBank ID: X15545

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: M13F
• 3′ sequencing primer: M13R

Terms and Licenses

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

Depositor Comments

As pMT316 but with the TGT codons for the two cysteines of barstar, C40 and C82, changed to GCG for alanine. The protein product, barstar A, has no sulfhydryl groups but is functional in vivo and in vitro, with only slight reductions in barnase binding and stability.