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URL: http://www.addgene.org/40942
Proper Citation: RRID:Addgene_40942
Insert Name: GFP-LacI**
Organism: E. coli
Bacterial Resistance: Ampicillin
Defining Citation: PMID:21724830
Vector Backbone Description: Backbone Marker:Andrew W. Murray Lab, UCSF (Straight, Sedat, & Murray, 1998. PMID: 9813090); Backbone Size:5650; Vector Backbone:pAFS135; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Comments: The plasmid pAFS135 (Straight, Sedat, and Murray, 1998) contains GFP(S65T, V163A)-LacI(P3Y, del C-terminal 11 aa)-SV40 NLS under the control of a HIS3 promoter fragment. To construct pAT123, the HIS3 marker of AFS135 was replaced with a ScaI-DraIII fragment from RS405 containing LEU2 (a kind gift from Karine Dubrana and Susan Gasser). The plasmid pAT222 was obtained by site-directed mutagenesis of pAT123 to create GFP-LacI**, using the primers (base mutations in lowercase): 5’- GTGGCACAgCAACTGGCGGaCAAAggtggcggaTCGTTGCTGATTGGCGTTGCCtCCT-3’ and 5’- AGGaGGCAACGCCAATCAGCAACGAtccgccaccTTTGtCCGCCAGTTGcTGTGCCAC- 3’. Alternate plasmid name: pAT123LacI2G(4mutations)
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Source: Addgene
