pcDNA-4mtD3cpv
(Plasmid #36324)

• Depositing Labs: Amy Palmer, Roger Tsien
• Publication: Palmer et al Chem Biol. 2006 May;13(5):521-30.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5400
• Total vector size (bp): 7800
• Modifications to backbone: mitochondrial targeting sequence N-terminal to insert
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH10B
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: 4mtD3cpv second generation cameleon (calcium sensor) targeted to mitochondria
• Alt name: D family cameleon
• Species: Synthetic
• Insert Size (bp): 2370
• GenBank ID: DQ479429.1
• Promoter: CMV
• Tag / Fusion Protein:
  • First 29 aa of human COX subunit 8a (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: T7 Forward
• 3′ sequencing primer: BGH Reverse

Resource Information

• Articles Citing this Plasmid:
  • 14 References

Terms and Licenses

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

Depositor Comments

Genetically encoded, ratiometric, calcium biosensor that contains enhanced CFP and circularly permuted Venus 173. D3cpv does not bind wild-type calmodulin. It has a Kd' of 0.6 uM.

See also Palmer & Tsien (2006) Nature Protocols.

The small discrepancy between the Addgene and depositor's sequence should not alter the function of the plasmid.