pET-BiFC
(Plasmid #87856)

• Purpose: pETDUET-1 based vector for use in BiMolecular Fluorescence Complementation based assays. MCS at 5' of each BiFC fragment.
• Depositing Lab: Dan Mulvihill
• Publication: Eastwood et al FEBS Lett. 2017 Feb 18. doi: 10.1002/1873-3468.12597.

Backbone

• Vector backbone: pETDUET-1
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5420
• Total vector size (bp): 6148
• Modifications to backbone: MCS & C-terminal Venus based fragment ligated between NcoI and BamHI sites. MCS & N-terminal Venus based fragment ligated between NdeI and BglII sites.
• Vector type: Bacterial 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 1

• Gene/Insert name: Encodes for MCS & C-term half of mVenus
• Alt name: VC155
• Species: Synthetic
• Insert Size (bp): 285
• Promoter: T7
• Tag / Fusion Protein:
  • BiFC C-terminal fragment (C terminal on backbone)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (not destroyed)
• 3′ cloning site: BamHI (destroyed during cloning)
• 5′ sequencing primer: pET Unpstream (Novagen primer 69214-3)
• 3′ sequencing primer: DuetDOWN1 Primer (Novagen primer 71179-3)

Gene/Insert 2

• Gene/Insert name: Encodes for MCS & N-term half of mVenus
• Alt name: VN155
• Species: Synthetic
• Insert Size (bp): 495
• Promoter: T7
• Tag / Fusion Protein:
  • BiFC C-terminal fragment (N terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: BglII (destroyed during cloning)
• 5′ sequencing primer: DUETUP2 Primer (Novagen primer 71180-3)
• 3′ sequencing primer: T7 Terminator primer

Resource Information

• A portion of this plasmid was derived from a plasmid made by: BiFC fragment DNA was generated synthetically to generate peptides described in: Y. Kodama, C.-D. Hu, An improved bimolecular fluorescence complementation assay with a high signal-to-noise ratio, BioTechniques. 49 (2010) 793–805. doi:10.2144/000113519.

Terms and Licenses

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