pcDNA-mito-ZifCV1.173
(Plasmid #59010)

• Purpose: Mitochondrial Zn-binding FRET biosensor
• Depositing Lab: Amy Palmer
• Publication: Park et al ACS Chem Biol. 2012 Aug 10.

Backbone

• Vector backbone: pcDNA3.1(+)
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5500
• Total vector size (bp): 7400
• Vector type: Mammalian Expression
• 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: mito-ZifCV1.173 genetically encoded Zn(II) sensor
• Species: Synthetic
• Insert Size (bp): 1926
• Promoter: CMV
• Tag / Fusion Protein:
  • See comment (N terminal on insert)

Cloning Information

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

Terms and Licenses

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

Depositor Comments

Genetically encoded, ratiometric, fluorescent biosensor targeted to the mitochondria. mito-ZifCV1.173 contains 4 tandem repeats of the first 29 aa of human cox subunit 8a (MSVLTPLLLRGLTGSARRLPVPRAKIHSL). HindIII restriction sites are at the 5' and 3' ends of the mitochondrial targeting sequence. Zn(II)-binding domain derived from mammalian Zif268. Contains truncated CFP and circularly permuted Venus 173.