pLL3.7 FLAG-YAP1-TEAD-P-H2B-mCherry
(Plasmid #128327)

• Purpose: Reporter to evaluate YAP1/TEAD-mediated gene transcription
• Depositing Lab: Yutaka Hata
• Publication: Maruyama et al Mol Cancer Res. 2018 Feb;16(2):197-211. doi: 10.1158/1541-7786.MCR-17-0382. Epub 2017 Oct 23.

Backbone

• Vector backbone: pLL3.7
• Modifications to backbone: Please refer to the paper.
• Vector type: Mammalian Expression, Lentiviral

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: H2B mCherry
• Species: H. sapiens (human)

Gene/Insert 2

• Gene/Insert name: YAP1
• Species: H. sapiens (human)
• Entrez Gene: YAP1 (a.k.a. COB1, YAP, YAP-1, YAP2, YAP65, YKI)
• Tag / Fusion Protein:
  • FLAG (N terminal on backbone)

Resource Information

• A portion of this plasmid was derived from a plasmid made by: H2B-mCherry from Robert Benezra (Addgene plasmid #20972)
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

pCIneoFH-YAP1 was digested with NotI, filled in, and cut with NheI. The isolated fragment was ligated into NheI/SmaI sites of pLL3.7 K122 to generate pLL3.7 K122 FH-YAP1. NcoI/BglII from pEYFP-C1 (Clontech Laboratories) was ligated into NcoI/BamHI of pGL3 to replace luciferase with YFP and to generate YFP reporter. BamHI/Sall and Sall/HindIII fragments from 8xGT-IIC-d51LucII luciferase reporter (a gift from Hiroshi Sasaki; RIKEN RDB08067) were ligated into BglII/HindIII sites of the YFP reporter to generate pTEAD-responsive-promoter YFP, which was digested with NotI and partially digested with HindIII. The resulting fragment and HindIII/XbaI fragment from H2B-mCherry (a gift from Robert Benezra; Addgene plasmid #20972) were ligated into NotI/XbaI sites of pLL3.7 K122 FH-YAP1 and pLL3.7 K122 to obtain pLL3.7 K122 FH-YAP1-ires-GFP-TEAD-responsive-promoter-H2B-mCherry reporter