AAV phSyn1-FSF-FLEX-ChR2(H134R)-EYFP-WPRE-bGHpA
(Plasmid #65454)

• Purpose: Can be used to generate AAV virus that will express the ChR2(H134R)-EYFP channelrhodopsin fusion protein in neurons under intersectional control by Flp and Cre recombinases
• Depositing Lab: Hongkui Zeng
• Publication: Madisen et al Neuron. 2015 Mar 4;85(5):942-58. doi: 10.1016/j.neuron.2015.02.022.

Backbone

• Vector backbone: pAAV-MCS
• Backbone manufacturer: Stratagene
• Modifications to backbone: A fragment containing: the hSyn1 promoter-a (Frt-Stop-Frt) cassette-a FLEX cassette with Lox71/LoxJTZ17 sites flanking (ChR2H134R-EYFP)-WPRE-bGHpA were swapped in to replace the CMV promoter-MCS-hGHpA sequence. Total insert size including the two ITRs is 4443 bp.
• Vector type: AAV

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Stbl2
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: ChR2(H134R)-EYFP
• Alt name: channelrhodopsin-2 variant H134R-EYFP
• Species: C. reinhardtii
• Insert Size (bp): 1671
• Mutation: H134R variant
• Promoter: hSyn1
• Tag / Fusion Protein:
  • EYFP (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Not1 (destroyed during cloning)
• 3′ cloning site: Not1 (destroyed during cloning)
• 5′ sequencing primer: none
• 3′ sequencing primer: none

Terms and Licenses

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

Depositor Comments

This plasmid has been found to be prone to multimerization. Plasmid multimerization often does not impact plasmid function, but may reduce transformation efficiencies. You may need to screen multiple colonies to isolate the monomeric version of this plasmid. If you still have trouble isolating the monomeric version, you might consider linearizing, gel extracting, re-ligating, and transforming the plasmid.