pAAV-EF1α-DIO-GtACR2-P2A-Voltron2ST-W3SL
(Plasmid #217676)

• Purpose: AAV-mediated, Cre-dependent co-expression of GtACR2 and soma-targeted Voltron2 (ORCHID) for assessing inhibitory receptor driving force through voltage imaging with concurrent activation of GtACR2.
• Depositing Lab: Joseph Raimondo
• Publication: Selfe et al bioRxiv 2023.08.30.555464

Backbone

• Vector backbone: pAAV
• Backbone size w/o insert (bp): 4158
• Total vector size (bp): 6951
• Vector type: Mammalian Expression, AAV

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: GtACR2-P2A-Voltron2ST
• Alt name: ORCHID (all-Optical Reporting of CHloride Ion Driving force)
• Species: Synthetic
• Insert Size (bp): 2793
• Promoter: EF‐1α
• Tag / Fusion Protein:
  • Soma targeting sequence (Kv2.1) on Voltron2 gene only. (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsrGI (unknown if destroyed)
• 3′ cloning site: Nhe1 (unknown if destroyed)
• 5′ sequencing primer: CTCCTCATGTCAATCACTCC; GTCCATGTATTCTTCGACCAG; GTGCAACATGCGGGATGATG; GTATGCGCTCTTCAGCTCTG; CAGCCACTTCATGTACTCGG
• 3′ sequencing primer: GTAATCCAGAGGTTGATTATC

Resource Information

• Supplemental Documents:
  • pAAV-EF1α-DIO-GtACR2-P2A-Voltron2ST-W3SL.gb
• A portion of this plasmid was derived from a plasmid made by: The Voltron2-ST gene originates from pGP-pcDNA3.1 Puro-CAG-Voltron2-ST (Addgene #172910), and was kindly provided by Eric Schreiter and Ahmed Abdelfattah. The GtACR2 gene originates from pAAV-EF1α-FRT-FLEX-GtACR2-EYFP (Addgene #114369).

Terms and Licenses

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

Depositor Comments

Please visit https://www.biorxiv.org/content/10.1101/2023.08.30.555464v2 for bioRxiv preprint.