pOTTC292 - pAAV EF1a floxed hChR2(H134R)-EYFP
(Plasmid #50834)

• Purpose: An AAV packaging vector that expresses channel rhodopsin 2 (H134R) (fused to EYFP) under the EF1a promoter, which can be deactivated (deleted) by recombination between the flanking loxP sites.
• Depositing Lab: Brandon Harvey
• Publication: McDevitt et al Cell Rep. 2014 Sep 25;8(6):1857-1869. doi: 10.1016/j.celrep.2014.08.037. Epub 2014 Sep 18.

Backbone

• Vector backbone: Addgene 20298
• Backbone manufacturer: Karl Deisseroth
• Backbone size w/o insert (bp): 5500
• Total vector size (bp): 7266
• Modifications to backbone: Removed double floxed inverted open reading frame (hChR2-EYFP) using KpnI and EcoRI digestion. Used In-Fusion cloning to recombine backbone with PCR-based insert (KpnI-loxP-hChR2-EYFP-loxP-EcoRI).
• Vector type: Mammalian Expression, AAV, Cre/Lox

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: hChR2 (H134R)
• Alt name: Channel Rhodopsin 2
• Species: C. reinhardtii
• Mutation: H134R
• GenBank ID: EF474017
• Promoter: EF1a
• Tag / Fusion Protein:
  • EYFP (C terminal on insert)

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: EF1a_F1
• 3′ sequencing primer: WPRE_R1

Resource Information

• Supplemental Documents:
  • pOTTC292 - GenBank format
• A portion of this plasmid was derived from a plasmid made by: The ORF for hChR2-EYFP, and the AAV vector backbone was obtained from Karl Deisseroth, but is also available from Addgene (Plasmid 20298).

Terms and Licenses

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