pEYFPC1-ECFP/GgVcl 1-419 control 1 FRET
|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 size w/o insert (bp) 4700
- Total vector size (bp) 6732
Vector typeMammalian Expression
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Gene/Insert nameECFP + Vcl 1-419
SpeciesG. gallus (chicken)
- Promoter CMV
/ Fusion Protein
- EYFP (N terminal on backbone)
- 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)
Terms and Licenses
- Not Available to Industry
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.
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