pET15b/GgVcl V1-851 (aka Vh)
(Plasmid #46172)

• 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: pET-15b
• Backbone manufacturer: EMD
• Backbone size w/o insert (bp): 5700
• Total vector size (bp): 8264
• 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

• Gene/Insert name: Vcl 1-851
• Alt name: Vinculin
• Alt name: Vcl
• Species: G. gallus (chicken)
• Insert Size (bp): 2559
• Mutation: aa 1-851 (aka Vh) head domain
• GenBank ID: NM_205441.1
• Entrez Gene: VCL (a.k.a. VINC1)
• Promoter: T7
• Tag / Fusion Protein:
  • His (N terminal on backbone)

Cloning Information

• Cloning method: TOPO Cloning
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7 terminal

Terms and Licenses

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

Depositor Comments

Using in-frame primers containing 5′-NdeI and 3′-XhoI sites, appropriate regions of the chicken vinculin cDNA sequence were amplified by PCR followed by addition of 3′-adenosine overhangs with Taq polymerase. The PCR products were ligated into the TOPO II plasmid (Invitrogen) and then subcloned into pET15b His tag expression vector (Novagen, Madison, WI) using NdeI and XhoI digestion of the multiple cloning site.