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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 (RRID:SCR_002037)
Description: Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.
URL: http://www.addgene.org
The NIDDK Information Network (dkNET) serves the needs of basic and clinical investigators by providing seamless access to large pools of data and research resources relevant to the mission of The National Institute of Diabetes Digestive and Kidney Diseases (NIDDK). dkNET is hosted at University of California San Diego, and is supported by NIH NIDDK grant U24DK097771.
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