URL: http://www.addgene.org/40592
Proper Citation: RRID:Addgene_40592
Insert Name: GFP-S65T
Organism: Homo sapiens
Bacterial Resistance: Ampicillin
Defining Citation: PMID:9224672
Vector Backbone Description: Backbone Marker:Michael Brenner (PMID: 7952266); Backbone Size:5500; Vector Backbone:pgfa2-lac2; Vector Types:Mammalian Expression, Bacterial Expression, Mouse Targeting; Bacterial Resistance:Ampicillin
Comments: This plasmid encodes a transgene to express hGFP-S65T under the control of the hGFAP promoter in transgenic mice. The plasmid Gfa2-lac2, containing the human GFAP promoter and mouse protamine 1 intron and polyadenylation signal on the 3' end, was kindly provided by Dr. M. Brenner of the NIH. The plasmid phGFP-S65T containing the mutated, humanized GFP cDNA was purchased from the Clonetech Laboratories. The transgene was constructed by excising the lacZ coding region from the pGfa2-lac2 plasmid by BamHI digestion, and replacing it by blunt end ligation with a 0.75-kb HindIII-XbaI fragment containing the entire GFP coding region from phGFP-S65T. hGFP-S65T is a variant of the Aequorea victoria green fluorescent protein that has been optimized for brighter fluorescence and higher expression in human (and mammalian) cells. hGFP-S65T contains more than 190 silent base changes to optimize the coding sequence based on human codon-usage preferences. The S65T mutation results in a single, red-shifted excitation peak at 490 nm and brighter fluorescence.
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Source: Addgene