VBS mutant of alpha catenin sensor
(Plasmid #71710)

• Purpose: This form of the alpha catenin conformation sensor lacks the vinculin binding site and does not bind vinculin.
• Depositing Lab: Deborah Leckband
• Publication: Kim et al Curr Biol. 2015 Jan 19;25(2):218-24. doi: 10.1016/j.cub.2014.11.017. Epub 2014 Dec 24.

Backbone

• Vector backbone: pCDNA3.1
• Backbone manufacturer: Invitrogen (Life Technologies)
• Backbone size w/o insert (bp): 5400
• Total vector size (bp): 9800
• 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): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: alpha-catenin ΔVBS FRET Sensor
• Species: H. sapiens (human)
• Insert Size (bp): 4400
• Mutation: Deleted amino acid residues 316-405 in α -catenin and substituted with the homologous sequence from mouse vinculin (aa 514-606)
• Promoter: CMV
• Tags / Fusion Proteins:
  • ECFP (N terminal on insert)
  • YPET (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AgeI (unknown if destroyed)
• 3′ cloning site: KpnI (unknown if destroyed)
• 5′ sequencing primer: CMV-F

Resource Information

• Supplemental Documents:
  • sensor construction protocol
• A portion of this plasmid was derived from a plasmid made by: acat 1-906 from David Rimm (Addgene plasmid # 24194)

Terms and Licenses

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

Depositor Comments

This plasmid is a derived from the alpha catenin conformation sensor whose Vinculin binding site has been substituted with the homologous sequence from mouse vinculin (aa 514-606)