pAAV-Syn-FLEX-rc [ChrimsonR-GFP]
(Plasmid #84480)

• Purpose: AAV-mediated expression of ChrimsonR-GFP under the Syn promoter, in floxed/reversed (Cre-dependent) manner. Using bGHpA signal.
• Depositing Lab: Edward Boyden
• Publication: Klapoetke et al Nat Methods. 2014 Mar;11(3):338-46. doi: 10.1038/nmeth.2836. Epub 2014 Feb 9.

Backbone

• Vector backbone: AAV-Syn-FLEX
• Backbone manufacturer: original from Stratagene
• Backbone size w/o insert (bp): 4684
• Total vector size (bp): 6469
• Vector type: Mammalian Expression, AAV

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Growth instructions: Can grow in DH5alpha at 37°C or Stbl3 at 30°C. Carbenicillin is preferred over ampicillin. In DH5alpha this plasmid may act more like a high copy plasmid, although in Stbl3 it may act more like a low copy plasmid.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: ChrimsonR-GFP
• Alt name: Chrimson K176R mutant-GFP
• Alt name: ChR88m19-GFP
• Alt name: Chlamydomonas noctigama channelrhodopsin K176R mutant-GFP
• Species: Chlamydomonas noctigama
• Insert Size (bp): 1785
• Mutation: Chrimson K176R mutant
• GenBank ID: KF992060
• Promoter: human synapsin promoter
• Tag / Fusion Protein:
  • GFP (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: KpnI (not destroyed)
• 5′ sequencing primer: gcacgggcgcgaccatctgc
• 3′ sequencing primer: TAGCGTAAAAGGAGCAACATAG

Terms and Licenses

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

Depositor Comments

The plasmid is fully sequenced in the coding sequence regions (opsin-fluorophore and important flanking regions). Multiple digestions were done to verify the vector structure. The construct and the virus were both tested in vitro.