pEGFPC1/GgVcl 1-1066 T12 mutant
(Plasmid #46266)

• Depositing Lab: Susan Craig
• Publication: Cohen et al J Biol Chem. 2005 Apr 29;280(17):17109-17. Epub 2005 Feb 22.

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 T12 mutant
• Alt name: Vinculin
• Alt name: Vcl
• Species: G. gallus (chicken)
• Insert Size (bp): 3259
• Mutation: T12 mutant (D974;K975;R976; R978A) autoinhibition decreased by 100x
• 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

Resource Information

• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

T12 mutation introduced by QuikChange PCR to Addgene plasmid 46265.

Mutant inhibits vinculin autoinhibition by ~ 100 fold, vinculin is partially activated and will bind talin, but not actin (unless both talin and actin are present at same time).