pAAV-hSyn-DIO {hCAR}off-{ChETA-EYFP}on-W3SL
(Plasmid #111392)

• Purpose: AAV vector with hSynapsin promoter, Cre-OFF hCAR (for efficient CAV-2 infection), Cre-ON ChETA-EYFP (for optogenetic activation), and W3SL regulatory cassette (for maximize cloning capacity)
• Depositing Lab: Adam Kepecs
• Publication: Li et al Neuron. 2018 Jun 6;98(5):905-917.e5. doi: 10.1016/j.neuron.2018.05.028.

Backbone

• Vector backbone: pAAV
• Backbone size w/o insert (bp): 4115
• Total vector size (bp): 6868
• Vector type: Mammalian Expression, AAV

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: hCAR
• Alt name: human CAR variant 1
• Species: H. sapiens (human)
• Insert Size (bp): 1095
• GenBank ID: NM_001338
• Entrez Gene: CXADR (a.k.a. CAR, CAR4/6, HCAR)
• Promoter: hSynapsin
• Tag / Fusion Protein:
  • Myc (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: AscI (not destroyed)
• 3′ cloning site: PacI (not destroyed)
• 5′ sequencing primer: CCAAATTGCGCATCCCCTATC

Gene/Insert 2

• Gene/Insert name: ChETA-EYFP
• Alt name: hChR2(E123T/H134R)-EYFP
• Species: Synthetic
• Insert Size (bp): 1659
• Promoter: hSynapsin
• Tag / Fusion Protein:
  • EYFP (C terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: PacI (not destroyed)
• 3′ cloning site: NheI (not destroyed)
• 5′ sequencing primer: CACGAACTCCAGCAGGACCATG
• 3′ sequencing primer: GCAGCGTATCCACATAGCG

Resource Information

• Supplemental Documents:
  • S23-pAAV-hSynDIO-hCARmyc-ChetaEYFP-W3SL.gb
• A portion of this plasmid was derived from a plasmid made by: Backbone comes from plasmid pAAV-Ef1a-DIO-{ChETA-EYFP} (Addgene, Plasmid #26968); W3SL cassette comes from plasmid pAAV-CW3SL-EGFP (Addgene, Plasmid #61463).

Terms and Licenses

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