pEGFPC1/GgVcl 1-1066 A50I K1306A, R1308A mutant
(Plasmid #46267)

• Depositing Lab: Susan Craig
• Publication: Craig lab unpublished plasmids (unpublished)

Backbone

• Vector backbone: pEGFP-C1
• Backbone manufacturer: Clonetech
• Backbone size w/o insert (bp): 4700
• Total vector size (bp): 7990
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Vcl
• Alt name: Vinculin
• Species: G. gallus (chicken)
• Insert Size (bp): 3259
• Mutation: K1306A, R1308A, A50I mutation; talin binding deficient
• GenBank ID: NM_205441.1
• Entrez Gene: VCL (a.k.a. VINC1)
• Promoter: CMV
• Tag / Fusion Protein:
  • EGFP (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: EGFP-C
• 3′ sequencing primer: SV40pA-R

Terms and Licenses

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

Depositor Comments

Chicken vinculin, full length(EcoRI fragment of p1005 subcloned into EcoRI-cut pEGFP-C1); A50I mutation (alanine 50 to isoleucine) introduced by QCPCR.
This mutant binds poorly (undetectably to talin in pull down assays; sensitivity of assay Kd= 10-20 uM).