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pcDNA3.1 V5 His-TOPO-GluCl optalpha-mYFP L9'F (Opt α-GluClv2.0)
(Plasmid #47387)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 47387 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pcDNA3.1/V5-His TOPO
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 5523
  • Total vector size (bp) 7644
  • Modifications to backbone
    Mutagenesis—Codon optimized sequences of the C. elegans GluCl channel, including optGluCl WT, optGluCl WT, optGluCl -XFP, and optGluCl -XFP (4), cloned into plasmid vector pcDNA3.1/V5-His TOPO (catalog no. K4800-01, Invitrogen) were used in this study. Fluorescent protein insertions (XFP) include enhanced yellow (YFP) and cyan (CFP) variants and are located in the M3-M4 loop (3). For convenience, the “opt” nomenclature is omitted throughout most of this paper. Mutations were made using the QuikChange II XL site-directed mutagenesis kit (catalog no. 200522, Agilent Technologies) and PfuTurboDNApolymerase (catalog no. 600250, Agilent Technologies
  • Vector type
    Mammalian Expression
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    Top10
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    glutamate-gated chloride Channels
  • Alt name
    GluCl optalpha-mYFP L9'F
  • Species
    C. elegans (nematode)
  • Insert Size (bp)
    2121
  • Mutation
    Leu9 prime mutated to F (Phenylalanine in M3-M4 loop, L315F); monomerizing A206K in YFP
  • GenBank ID
    AY273891.1
  • Promoter CMV

Cloning Information

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.
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:

    pcDNA3.1 V5 His-TOPO-GluCl optalpha-mYFP L9'F (Opt α-GluClv2.0) was a gift from Henry Lester (Addgene plasmid # 47387 ; http://n2t.net/addgene:47387 ; RRID:Addgene_47387)
  • For your References section:

    An Engineered GluCl Channel for Sensitive, Consistent Neuronal Silencing by Ivermectin. Frazier SJ, Cohen BN, Lester HA. J Biol Chem. 2013 May 29. 10.1074/jbc.M112.423921 PubMed 23720773