AAVS1 CAG rtTA3 TauWT 2N4R-EGFP
(Plasmid #132389)

• Purpose: Exchange cassette containing inducible TauWT 2N4R-EGFP fusion, CAG-driven rtTA3, Puromycin resistance marker, targets AAVS1 site
• Depositing Lab: Gerold Schmitt-Ulms
• Publication: Wang et al Bio Protoc. 2020 May 5;10(9):e3615. doi: 10.21769/BioProtoc.3615. eCollection 2020 May 5.

Backbone

• Vector backbone: AAVS1-T2A-Puro-Cas9-TREtight
• Backbone manufacturer: Addgene
• Total vector size (bp): 8048
• Vector type: Mammalian Expression, Cre/Lox, CRISPR
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Bacteria may truncate DNA, please select multiple clones and perform restriction digest after purification to confirm.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: human TauWT 2N4R-EGFP
• Species: H. sapiens (human), Synthetic
• Entrez Gene: MAPT (a.k.a. DDPAC, FTD1, FTDP-17, MAPTL, MSTD, MTBT1, MTBT2, PPND, PPP1R103, TAU, Tau-PHF6, tau-40)
• Promoter: TREtight
• Tag / Fusion Protein:
  • EGFP (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BbvCI (not destroyed)
• 3′ cloning site: KpnI (not destroyed)
• 5′ sequencing primer: TTCAGGTTGGACCGGTCCTCA

Resource Information

• Supplemental Documents:
  • AAVS1 CAG rtTA3 TauWT 2N4R-EGFP

Terms and Licenses

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

Depositor Comments

Please note: Plasmid contains a 55 basepair deletion in the poly-GC track of the CAG promoter. It is unknown if this deletion affects plasmid function, but depositing lab was able to properly express proteins from the plasmid.