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URL: http://www.addgene.org/46035
Proper Citation: RRID:Addgene_46035
Insert Name: WRN K577M
Organism: Homo sapiens
Bacterial Resistance: Ampicillin
Defining Citation: PMID:15084309
Vector Backbone Description: Backbone Marker:Clontech; Vector Backbone:pBI; Vector Types:Mammalian Expression, TetOn, bidirectional promoter; Bacterial Resistance:Ampicillin
Comments: Bidirectional vector expresses WRN and GFP at similar expression levels. In the associated article, cells were transiently transfected with this plasmid together with a tTA transactivator plasmid (Gossen, 1992). This is a mammalian expression vector for helicase-dead human WRN protein. It was constructed from pBI, a Clontech plasmid with a bi-directional promoter+ TRE backbone, modified by insertion of a polylinker containing BspD1 and Xhol sites followed by insertion of a myc-WRN K577M ORF- created by making K577M (A>T) mutation in pMM290 sequence. The S2G mutation was a result of the cloning strategy to add the myc tag. The WRN protein containing S2G was biochemically verified and is wild-type in terms of activities, as described in the associated publication.
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Source: Addgene
