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pEGFPC1/GgVcl 1-258 (aka VD1)
(Plasmid #46270)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 46270 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pEGFP-C1
  • Backbone manufacturer
    Clonetech
  • Backbone size w/o insert (bp) 4700
  • Total vector size (bp) 7316
  • Modifications to backbone
    vector modified to contain unique Nde, Xho sites.
  • Vector type
    Mammalian Expression
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    Vcl 1-258 (aka VD1)
  • Alt name
    Vinculin
  • Alt name
    Vcl
  • Species
    G. gallus (chicken)
  • Insert Size (bp)
    774
  • Mutation
    1-258 (VD1 domain)
  • GenBank ID
    NM_205441.1
  • Entrez Gene
    VCL
  • Promoter CMV
  • Tag / Fusion Protein
    • EGFP (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NdeI (not destroyed)
  • 3′ cloning site XhoI (not destroyed)
  • 5′ sequencing primer EGFP-C
  • 3′ sequencing primer SV40pA-R
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Nde-Xho fragment from pET15b/Vh1-258 (Addgene plasmid 46173) was subcloned into EGFP/V1-1066 (Addgene plasmid 46265) vector modified to contain unique Nde, Xho sites. Modifications were made by QuikChange PCR to facilitate cloning of cDNAs previously in the pET15b vector. The NdeI site present in the cytomegalovirus promoter region was removed (mutating adenine 234 to thymine), and
a new NdeI site was introduced into the multiple cloning site immediately before the vinculin start codon. Additionally, the 5-XhoI site was removed (mutating cytosine 1345 to adenine), and a 3-XhoI site was introduced 3' to the vinculin cDNA in lieu of a PstI site.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pEGFPC1/GgVcl 1-258 (aka VD1) was a gift from Susan Craig (Addgene plasmid # 46270 ; http://n2t.net/addgene:46270 ; RRID:Addgene_46270)
  • For your References section:

    A conformational switch in vinculin drives formation and dynamics of a talin-vinculin complex at focal adhesions. Cohen DM, Kutscher B, Chen H, Murphy DB, Craig SW. J Biol Chem. 2006 Jun 9;281(23):16006-15. Epub 2006 Apr 10. 10.1074/jbc.M600738200 PubMed 16608855