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

• Purpose: allow pharmacologically induced silencing of electrical activity in CNS neurons upon exposure to the anthelmintic drug ivermectin
• Depositing Lab: Henry Lester
• Publication: Frazier et al J Biol Chem. 2013 May 29.

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

• Cloning method: TOPO Cloning
• 5′ sequencing primer: CMV-F; T7
• 3′ sequencing primer: BGH-Rev

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry