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pEYFPC1-ECFP/GgVcl 1-419 control 1 FRET
(Plasmid #46277)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 46277 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pEYFP-C1
  • Backbone manufacturer
    Clonetech
  • Backbone size w/o insert (bp) 4700
  • Total vector size (bp) 6732
  • 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
    ECFP + Vcl 1-419
  • Alt name
    Vinculin
  • Alt name
    Vcl
  • Species
    G. gallus (chicken)
  • Mutation
    aa 1-419
  • Entrez Gene
    VCL
  • Promoter CMV
  • Tag / Fusion Protein
    • EYFP (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site HindIII (not destroyed)
  • 3′ cloning site KpnI (not destroyed)
  • 5′ sequencing primer CMV-F
  • 3′ sequencing primer SV40pA-R
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

To make the control FRET probe pEYFP-C1/CFPV1-400, a HindIII site was engineered before the ATG site of pET15b/CFP-V1-851 and a KpnI site after codon 400 by PCR amplification with 5 -CAAGCTTCGatggtgagcaagggc-3 and 5 -GGTACCTCAtgcaactttccttgc-3 . The PCR product was introduced into TOPO pCRII and sequenced. The HindIII–KpnI fragment of CFP-vinculin1-400 was subcloned into pEYFP-C1 to generate the control plasmid pEYFP-CFPV1-400.

Tandem EYFP-ECFP- hooked to portion of vinculin that localizes to FA; maximum FRET.

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:

    pEYFPC1-ECFP/GgVcl 1-419 control 1 FRET was a gift from Susan Craig (Addgene plasmid # 46277 ; http://n2t.net/addgene:46277 ; RRID:Addgene_46277)
  • For your References section:

    Spatial distribution and functional significance of activated vinculin in living cells. Chen H, Cohen DM, Choudhury DM, Kioka N, Craig SW. J Cell Biol. 2005 May 9;169(3):459-70. 10.1083/jcb.200410100 PubMed 15883197