pAAV CAG-DO-TC66T-mCherry
(Plasmid #175178)

• Purpose: Cre-off TVA-mCherry fusion protein with 66T mutation
• Depositing Lab: Bernardo Sabatini
• Publication: Melzer et al Cell. 2021 Oct 28;184(22):5622-5634.e25. doi: 10.1016/j.cell.2021.09.013. Epub 2021 Oct 4.

Backbone

• Vector backbone: pAAV
• Total vector size (bp): 7238
• Modifications to backbone: double-floxed non-inverted open reading frame encoding TC66T-mCherry fusion protein driven by CAG promoter
• Vector type: AAV

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: TVA
• Alt name: CAG-DO-TC66T-mCherry
• Insert Size (bp): 3928
• Mutation: E66T
• Promoter: CAG
• Tag / Fusion Protein:
  • mCherry (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SalI (unknown if destroyed)
• 3′ cloning site: AscI (unknown if destroyed)
• 5′ sequencing primer: N/A
• 3′ sequencing primer: N/A

Resource Information

• A portion of this plasmid was derived from a plasmid made by: pAAV CAG-DO-TC66T-mCherry plasmid was generated by restriction enzyme digest of pAAV-CAG-Flex-TC66T with SalI and AscI to cut out the inverted reading frame encoding TC66T-mCherry. The endings of TC66T-mCherry were then mutated to reverse the AscI and SalI restriction enzyme sites and the new insert SalI-TC66T-mCherry-AscI ligated into the original vector as a non-inverted reading frame. The CAG-Flex-TC66T plasmid was a gift from Liqun Luo (Addgene plasmid # 48331) (Miyamichi et al., 2013).

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry