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pET15b/EYFP-GgVcl 1-258
(Plasmid #46263)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 46263 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pET-15b
  • Backbone manufacturer
    EMD
  • Backbone size w/o insert (bp) 5700
  • Total vector size (bp) 7238
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    Vcl 1-258
  • Alt name
    Vinculin
  • Species
    G. gallus (chicken)
  • Insert Size (bp)
    780
  • Mutation
    aa 1-258 (VD1)
  • GenBank ID
    NM_205441.1
  • Entrez Gene
    VCL
  • Promoter T7
  • Tags / Fusion Proteins
    • His (N terminal on backbone)
    • EYFP (N terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site Nde1 (not destroyed)
  • 3′ cloning site Xho1 (not destroyed)
  • 5′ sequencing primer T7
  • 3′ sequencing primer T7 terminal
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

EYFP was PCR amplified to introduce NdeI site at both 5' and 3' ends. The NdeI flanked EYFP was inserted into NdeI site of pET15b/Vh1-258 (Addgene plasmid 46173), where V1-258 was cloned between NdeI and XhoI. Plasmid contains a 12-residue spacer between the EYFP and vinculin domain.

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:

    pET15b/EYFP-GgVcl 1-258 was a gift from Susan Craig (Addgene plasmid # 46263 ; http://n2t.net/addgene:46263 ; RRID:Addgene_46263)
  • For your References section:

    Two distinct head-tail interfaces cooperate to suppress activation of vinculin by talin. Cohen DM, Chen H, Johnson RP, Choudhury B, Craig SW. J Biol Chem. 2005 Apr 29;280(17):17109-17. Epub 2005 Feb 22. 10.1074/jbc.M414704200 PubMed 15728584