URL: http://www.addgene.org/41732
Proper Citation: RRID:Addgene_41732
Insert Name: GltI based Glutamate Biosensor
Organism: Synthetic
Bacterial Resistance: Ampicillin
Defining Citation: PMID:23314171
Vector Backbone Description: Backbone Marker:modified from Invitrogen; Backbone Size:5300; Vector Backbone:pCMV(MinDis) [variant of pDisplay]; Vector Types:Mammalian Expression, Glutamate Biosensor; Bacterial Resistance:Ampicillin
Comments: ** A newer version of this sensor is available. Please see iGluSnFR3 plasmids at https://www.addgene.org/browse/article/28220233/ ** Biosensor iGluSnFRa is a single-wavelength glutamate sensor constructed from E. coli GltI and cpGFP. iGluSnFR is bright and photostable, with 4.5 (ΔF/F)max in vitro, under both 1- and 2-photon illumination. pCMV(MinDis).iGluSnFR was constructed by subcloning the GltI253-cpGFP.L1LV/L2NP gene from pRSET.GltI253-cpGFP.L1LV/L2NP (Addgene plasmid #41733) into the pDisplay vector (Invitrogen) between the BglII and PstI sites. The hemagglutinin (HA) tag was removed by PCR, generating the backbone variant dubbed pCMV(MinDis). This construct encodes an N-terminal mouse immunoglobin κ-chain leader sequence, which directs the protein to the secretory pathway; the GltI253.L1LV/L2NP protein; a myc epitope tag; and at the C-terminus, a platelet derived growth factor receptor transmembrane helix, which anchors the protein to the plasma membrane, displaying it on the extracellular side. Detailed amino acid sequences are provided in the supplementary documents available above. Alternate plasmid names: YbeJ253.L1LVL2NP.pMinDis; pMinDis.iGluSnFR These iGluSnFR constructs contain a K158R (GFP numbering) mutation that is not referenced in the publication. Depositing lab has confirmed that this mutation is expected and that the plasmids function as described.
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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