pBad-Int Set Flipper
(Plasmid #38209)

• Depositing Lab: Drew Endy
• Publication: Bonnet et al Proc Natl Acad Sci U S A. 2012 Jun 5;109(23):8884-9. Epub 2012 May 21.

Backbone

• Vector backbone: pSB3K1
• Vector type: Synthetic Biology ; BioBrick cloning vector

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Bxb1 Integrase
• Promoter: pBAD/AraC
• Tag / Fusion Protein:
  • His (N terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: pBAD-F

Terms and Licenses

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

Depositor Comments

The Bxb1 integrase was cloned downstream of the PBAD/AraC promoter (BBa_I0500) on pSB3K1 plasmid bearing a p15A origin of replication (15-20 copies). This version of the integrase has a 6-His-tag which we found to stabilize the protein. Therefore, a weak RBS (BBa_B0031) and a LAA ssrA tag was added to reduce the basal expression of the enzyme.