ρ1-EM GABAA receptor in pEZT-BM
(Plasmid #213072)

• Purpose: Express human GABA-A rho1 receptor with truncated intracellular domain and superfolder GFP insertion in ICD
• Depositing Lab: Erik Lindahl
• Publication: Cowgill et al Neuron. 2023 Nov 1;111(21):3450-3464.e5. doi: 10.1016/j.neuron.2023.08.006. Epub 2023 Sep 1.

Backbone

• Vector backbone: pEZT-BM
• Backbone manufacturer: Addgene Plasmid #74099
• Backbone size w/o insert (bp): 7436
• Total vector size (bp): 9494
• Vector type: Mammalian Expression ; Bacmam, Baculovirus, oocyte expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Human GABAa rho1 receptor
• Alt name: GABAa rho1
• Alt name: GABRR1
• Alt name: GABAC receptor
• Species: H. sapiens (human)
• Mutation: Encodes residues S58–L383 and D451–S479, separated by a sequence encoding the superfolder green fluorescent protein.81 Prior to the N-terminal truncation site, we added a membrane-translocation signal peptide from the human β3 GABAAR, a Twin-Strep tag, and a recognition site for tobacco etch virus for expression and purification purposes.
• Entrez Gene: GABRR1
• Promoter: CMV and T7
• Tags / Fusion Proteins:
  • superfolderGFP
  • Twin-Strep tag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (unknown if destroyed)
• 3′ cloning site: KpnI (unknown if destroyed)
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: WPRE Reverse

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Purchased from genscript

Terms and Licenses

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