BirA in pENTR1A (w860-1)
(Plasmid #29645)

• Depositing Lab: Eric Campeau
• Publication: latest plasmids (unpublished)

Backbone

• Vector backbone: pENTR1A
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 2266
• Vector type: Entry vector

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: TOP10F', 37oC
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: BirA
• Alt name: biotin ligase
• Insert Size (bp): 980

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Eco RI (not destroyed)
• 3′ cloning site: Eco RV (not destroyed)
• 5′ sequencing primer: ENTRforw
• 3′ sequencing primer: ENTRrev

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

The BirA gene was amplified from the E.coli TOP10F' strain, sequence confirmed, and functionally tested in E.coli using a protein fused to an AviTag.

Please note that using this plasmid in a LR reaction with a Destination vector encoding an N-terminal tag is not recommended as it would not be in frame with the BirA sequence. This plasmid was designed to be used to biotinylate substrates in vivo. To add a tag to BirA, clone the desired tag into the vector itself, rather than using Gateway cloning--this approach will also prevent unnecessary sequence in the expressed protein.