pAAV-EF1a-double floxed-ClopHensorN-WPRE-HGHpA
(Plasmid #193728)

• Purpose: Cre-activated AAV expression of ClopHensorN for measuring intracellular chloride and pH in genetically-defined cell types
• Depositing Lab: Colin Akerman
• Publication: Calin et al iScience. 2023 Mar 9;26(4):106363. doi: 10.1016/j.isci.2023.106363. eCollection 2023 Apr 21.

Backbone

• Vector backbone: pAAV-EF1a-double floxed-hChR2(H134R)-mCherry-WPRE-HGHpA (Addgene #20297)
• Backbone manufacturer: Karl Deisseroth
• Backbone size w/o insert (bp): 5601
• Total vector size (bp): 7842
• Modifications to backbone: Floxed ClopHensorN was generated by subcloning the inverted sequence of ClopHensorN (from Addgene plasmid # 50758) into the backbone of the double-floxed plasmid (Addgene plasmid # 20297), which had been prepared by removing the insert.
• 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: ClopHensorN
• Alt name: E2GFP fused to Tandem Tomato
• Species: Synthetic
• Insert Size (bp): 2264
• Promoter: EF1a
• Tag / Fusion Protein:
  • Strep-II (N terminal on insert) (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AscI (not destroyed)
• 3′ cloning site: NheI (not destroyed)
• 5′ sequencing primer: N/A
• 3′ sequencing primer: N/A

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry