FUW-tetO-wtYAP
(Plasmid #84009)

• Purpose: expresses wtYAP in mammalian cells under the control of a doxycycline-inducible promoter
• Depositing Lab: Stefano Piccolo
• Publication: Panciera et al Cell Stem Cell. 2016 Sep 9. pii: S1934-5909(16)30256-9. doi: 10.1016/j.stem.2016.08.009.

Backbone

• Vector backbone: FUW-tetO-MCS
• Backbone manufacturer: Piccolo lab
• Backbone size w/o insert (bp): 8500
• Total vector size (bp): 9970
• Vector type: Mammalian Expression, Lentiviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Growth instructions: Stbl3, 37ºC
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: YAP1 (siRNA insensitive)
• Species: H. sapiens (human)
• Insert Size (bp): 1540
• GenBank ID: NM_001130145.2
• Entrez Gene: YAP1 (a.k.a. COB1, YAP, YAP-1, YAP2, YAP65, YKI)
• Promoter: TRE-CMV
• Tag / Fusion Protein:
  • FLAG (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: PshAI (destroyed during cloning)
• 5′ sequencing primer: LNCX
• 3′ sequencing primer: pBluescriptKS

Resource Information

• Articles Citing this Plasmid:
  • 5 References

Terms and Licenses

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

Depositor Comments

The insert (FLAG-hYAP1 siRNA insensitive) was generated by PCR from original cDNA kindly provided by Dr. K.L. Guan, and made insensitive to YAP siRNA#1 and siRNA#2 by point mutagenesis (as in Aragona et al., 2013).

YAP siRNA#1 sequence: GACAUCUUCUGGUCAGAGA

YAP siRNA#2 sequence: CUGGUCAGAGAUACUUCUU