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URL: http://www.addgene.org/20409
Proper Citation: RRID:Addgene_20409
Insert Name: GFP
Organism: Aequorea victoria
Bacterial Resistance: Ampicillin
Defining Citation: PMID:8707053
Vector Backbone Description: Backbone Marker:Ezaz-Nikpay et al, 1994 Protein Science; Backbone Size:3531; Vector Backbone:pKEN; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: An XbaI site was introduced at the beginning of GFP, see attached map for more details. The protein is expressed from IPTG inducible promoter in pKEN vector. GFP is cloned as XbaI-PstI fragment into the polylinker; there is also NdeI site at the AUG that can be used. Mut 2 is probably best for expression in bacteria. Overexpression of GFP renders the cells osmosensitive. In the absence of IPTG, DH12 carrying the enclosed plasmids will lyse in LB. We recommend growing this plasmid in low salt LB (<5g/L NaCl); alternatively use E.coli strain that grossly overproduces lacI and totally shuts expression down.
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Source: Addgene
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