AAV-CAG-FLPX-rc [Jaws-KGC-tdTomato-ER2]
(Plasmid #195488)

• Purpose: AAV-mediated expression of Jaws-KGC-tdTomato-ER2 under the CAG promoter, in FLPX/reversed (Frt-dependent) manner.
• Depositing Lab: Edward Boyden
• Publication: Chuong et al Nat Neurosci. 2014 Aug;17(8):1123-9. doi: 10.1038/nn.3752. Epub 2014 Jul 6.

Backbone

• Vector backbone: AAV with CAG promter
• Backbone size w/o insert (bp): 4900
• Total vector size (bp): 7264
• 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 use 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: Jaws-KGC-tdTomato-ER2
• Alt name: Halo57-KGC-tdTomato-ER2
• Alt name: Jaws
• Species: Haloarcula salinarum (stain Shark)
• Insert Size (bp): 2364
• Mutation: K200R W214F
• GenBank ID: KM000926.1
• Promoter: CAG
• Tags / Fusion Proteins:
  • KGC (C terminal on insert)
  • tdTomato (C terminal on insert)
  • ER2 (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: ctagagcctctgctaacc
• 3′ sequencing primer: gcagcgtatccacatagcg

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• 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.