ACY
(Plasmid #12396)

• Depositing Lab: David Baker
• Publication: Dantas et al J Mol Biol. 2003 Sep 12. 332(2):449-60.

Backbone

• Vector backbone: pet29 b(+)
• Backbone size w/o insert (bp): 5371
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Plasmids are provided in DH5 alpha, but for protein expression, BL21(DE3)pLysS is recommended.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: ACY
• Alt name: acylphophatase
• Insert Size (bp): 390
• Mutation: RosettaDesign was used to design sequences. Alignment to wild-type can be seen in the article.
• Tag / Fusion Protein:
  • 6xHis (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: T7

Terms and Licenses

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

Depositor Comments

A synthetic gene which placed the sequence under the control of the T7 promoter, a C-terminal 6X His tag, and codon usage optimized for Escherichia coli was obtained from BlueHeron Biotechnologies. The gene was then subcloned into the pet29 b(+) vector from Novagen.