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GgVcl 1-1066 FRET tail probe 1
(Plasmid #46276)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 46276 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pECFP-N3
  • Backbone manufacturer
    Craig lab
  • Modifications to backbone
    pECFP-N3 vector was constructed as an intermediate vector for generating tail probe. ECFP was PCR amplified with 5 -GGTACCatggtgagcaagggc-3 and 5 -GCGGCCGCTttacttgtacagctc-3 to generate 5' KpnI and 3 ' NotI. The product was subcloned into TOPO pCRII and sequenced. The KpnI and NotI fragment of pCRII/ECFP was used to replace the EGFP fragment of pEGFP-N3 to generate pECFP-N3.
  • 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 1-1066 + EYFP
  • Alt name
    Vinculin
  • Alt name
    Vcl
  • Species
    G. gallus (chicken)
  • Mutation
    addition of 9 bp linker and EYFP at aa 883
  • Entrez Gene
    VCL
  • Promoter CMV
  • Tag / Fusion Protein
    • ECFP (C terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site EcoRI (not destroyed)
  • 3′ cloning site SalI (not destroyed)
  • 5′ sequencing primer CMV-F
  • 3′ sequencing primer ?
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

A 9-bp fragment encoding a NotI site was introduced by mutagenesis into pEGFP-C1/vinculin cDNA (chicken) immediately after the codon for aa 883 (Coutu and Craig, 1988) using
the Quick Change kit (Stratagene). The cDNA of EYFP (CLONTECH Laboratories, Inc.) minus the stop codon and flanked by NotI was inserted into NotI-digested vinculin.

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:

    GgVcl 1-1066 FRET tail probe 1 was a gift from Susan Craig (Addgene plasmid # 46276 ; http://n2t.net/addgene:46276 ; RRID:Addgene_46276)
  • 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