pETDUET-GST AO7REwt
(Plasmid #213935)

• Purpose: GST tagged AO7 encoding aa 126-258 for expression in bacteria
• Depositing Lab: Allan Weissman
• Publication: Li et al J Biol Chem. 2015 Dec 18;290(51):30225-39. doi: 10.1074/jbc.M115.685867. Epub 2015 Oct 16.

Backbone

• Vector backbone: pETDUET-1
• Backbone manufacturer: Millipore-Sigma
• Backbone size w/o insert (bp): 5420
• Modifications to backbone: An N-terminal GST fusion protein was cloned in place of the His6 tag (NcoI-BamHI). A PreScission Protease (GE Healthcare) sequence (LEVLFQGP) was included to enable removal of the GST.
• 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: AO7 aa 126-258
• Alt name: RNF25 AO7
• Species: H. sapiens (human)
• Insert Size (bp): 400
• Mutation: AO7 cloned into the first multiple cloning site of a modified pETDUET-1
• Entrez Gene: RNF25 (a.k.a. AO7)
• Promoter: T7 promoter-1
• Tag / Fusion Protein:
  • GST (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Bam1H1 (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: Unknown

Terms and Licenses

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

Depositor Comments

An N-terminal GST fusion protein was cloned in place of the His6 tag (NcoI-BamHI). A PreScission Protease (GE Healthcare) sequence (LEVLFQGP) was included to enable removal of the GST.