BRAC/pcDNA3
(Plasmid #51967)

• Purpose: mammalian expression of BRAC, a BRET-based autofluorescent Ca2+ indicator
• Depositing Lab: Takeharu Nagai
• Publication: Saito et al PLoS One. 2010 Apr 1;5(4):e9935. doi: 10.1371/journal.pone.0009935.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Vector type: Mammalian 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: BRAC
• Alt name: mVenus-Calmodulin-M13-Rluc8
• Species: Synthetic
• Insert Size (bp): 2200

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: BGHrev

Terms and Licenses

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

Depositor Comments

GenBank accession number: GU180352. BRAC is composed of the Ca2+-sensing domain, calmodulin (CaM) and M13, sandwiched between Venus and RLuc8. In the Ca2+ free state, CaM-M13 has an extended conformation so Venus is not located close to RLuc8 and thus only weak emission can be seen from Venus due to low BRET efficiency. Upon Ca2+ binding to CaM, Ca2+-CaM makes a compact complex with M13, which induces efficient BRET from RLuc8 to Venus resulting in fluorescence emission peaking at 530 nm.