pET-alpha-synuclein-BiFC
(Plasmid #87855)

• Purpose: pETDUET-1 based vector for BiMolecular Fluorescence Complementation based assays of alpha-synuclein oligomerisation
• 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): 6918
• 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: alpha-synuclein-VC155
• Species: H. sapiens (human), Synthetic
• Insert Size (bp): 686
• Promoter: T7
• Tag / Fusion Protein:
  • C-terminal fragment of mVenus BiFC protein (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (not destroyed)
• 3′ cloning site: BamH1 (destroyed during cloning)
• 5′ sequencing primer: pET Upstream
• 3′ sequencing primer: DuetDOWN1 Primer

Gene/Insert 2

• Gene/Insert name: alpha-synuclein-VN154
• Species: H. sapiens (human), Synthetic
• Insert Size (bp): 903
• Promoter: T7
• Tag / Fusion Protein:
  • N-terminal fragment of mVenus BiFC protein (N terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (destroyed during cloning)
• 3′ cloning site: BglII (destroyed during cloning)
• 5′ sequencing primer: DuetUP2 Primer
• 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