pET22b-EC12-129/138Avi
(Plasmid #47927)

• Purpose: Expresses E-cadherin EC12 with C9A, K129C/D138C mutations and avi tag in E.Coli.
• Depositing Lab: Tanja Kortemme
• Publication: Ritterson et al J Am Chem Soc. 2013 Aug 7.

Backbone

• Vector backbone: pET-22b(+)
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5406
• Total vector size (bp): 6117
• Modifications to backbone: None
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Top10
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: E-Cadherin EC12
• Alt name: EC12
• Species: M. musculus (mouse)
• Insert Size (bp): 750
• Mutation: Mutations C9A, K129C, D138C
• Promoter: T7
• Tags / Fusion Proteins:
  • 6xHis (N terminal on insert)
  • TEV Cleavage (N terminal on insert)
  • Avi Tag (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: taatacgactcactatagg
• 3′ sequencing primer: ctagttattgctcagcgg

Terms and Licenses

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

Depositor Comments

Plasmid contains E-Cadherin domains 1 and 2 (aa157-375, when compared to GenBank reference sequence NP_033994.1). Numbering of the mutations is relative to this sequence. The pro-peptide sequence that is normally cleaved during maturation was intentionally removed as part of the cloning strategy for expression in bacteria.