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pCMV(MinDis).iGluSnFR
(Plasmid #41732)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 41732 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pCMV(MinDis) [variant of pDisplay]
  • Backbone manufacturer
    modified from Invitrogen
  • Backbone size w/o insert (bp) 5300
  • Total vector size (bp) 6829
  • Modifications to backbone
    From pDisplay: Hemagglutinin (HA) tag removed
  • Vector type
    Mammalian Expression ; Glutamate Biosensor
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    GltI based Glutamate Biosensor
  • Alt name
    iGluSnFR
  • Alt name
    gltI/ybeJ
  • Alt name
    cpGFP146
  • Species
    Synthetic; E. coli
  • Insert Size (bp)
    1570
  • Mutation
    cpGFP146 variant from EcMBP165-cpGFP.PPYF.pRSET inserted between AA253-254 of E. coli gltI/ybeJ
  • Promoter CMV
  • Tags / Fusion Proteins
    • mouse Ig κ-chain (N terminal on backbone)
    • Myc (C terminal on backbone)
    • PDGFR transmembrane helix (aa 513-561) (C terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BglII (not destroyed)
  • 3′ cloning site PstI (not destroyed)
  • 5′ sequencing primer CMV-F
  • 3′ sequencing primer BGH-rev
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

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

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pCMV(MinDis).iGluSnFR was a gift from Loren Looger (Addgene plasmid # 41732)
  • For your References section:

    An optimized fluorescent probe for visualizing glutamate neurotransmission. Marvin JS, Borghuis BG, Tian L, Cichon J, Harnett MT, Akerboom J, Gordus A, Renninger SL, Chen TW, Bargmann CI, Orger MB, Schreiter ER, Demb JB, Gan WB, Hires SA, Looger LL. Nat Methods. 2013 Feb;10(2):162-70. doi: 10.1038/nmeth.2333. Epub 2013 Jan 13. 10.1038/nmeth.2333 PubMed 23314171