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
• Depositing Organization: California Institute of Technology
  Located in United States of America
• Publication: Frazier et al J Biol Chem. 2013 May 29.
• Citations: 1 Article

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

Terms and Licenses

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

DNA (Catalog # 47387-D50)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 50 μg
• Concentration: 1 μg/μL
• Pricing: $495 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

DNA (Catalog # 47387-D100)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 100 μg
• Concentration: 1 μg/μL
• Pricing: $585 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.