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URL: http://www.addgene.org/21871
Proper Citation: RRID:Addgene_21871
Bacterial Resistance: Ampicillin
Defining Citation: PMID:19798082
Vector Backbone Description: Backbone Size:3508; Vector Backbone:pMLM36; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: Combinatorial libraries of zinc finger arrays made by OPEN are cloned into this plasmid (see Maeder et al., Mol Cel 2008, PMID 18657511). Ligate zinc finger array fragments with BbsI-digested pBR-UV5-GP-FD2 vector backbone to create plasmids that express the zinc finger array as a FLAG-tagged Gal11P fusion in the B2H system. The lacUV5 promoter which drives expression of the Gal11P fusion protein contains a lac operator and therefore can be repressed by lac repressor. This plasmid is a low copy number plasmid and therefore we recommend that minipreps be performed from a 4 to 10 ml culture. Annotations for pMLM36: 12-17: lacUV5 promoter -35 box / 36-41: lacUV5 promoter -10 box / 48: lacUV5 promoter transcription start point / 86-88: Start codon for Gal11P protein fragment / 89-358: Coding sequence for Gal11P a.a. 263-352 (with additional translationally silent mutations) / 407-412: BbsI site #1 / 433-438: BbsI site #2 / 568-1024: f1 phage origin of replication / 1156-2013: beta-lactamase gene cassette (confers resistance to ampicillin and carbenicillin) / 2016-2956: ColE1 origin of replication (plasmid origin) / 3203-3390: rop gene (associated with regulation of copy number)
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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.
Email: info_at_ dknet.org
