Gateway-Based Expression Constructs

Expression constructs were generated with a N‐terminal Strep‐HA‐tagged bait proteins and entry clones of a Gateway compatible human clone collection (ORFeome v5.1 and v8.1). The integration of the entry clones into the Gateway destination vectors (pcDNA5/FRT/TO/SH/GW; Glatter et al, 2009 (link)) was performed with an enzymatic LR clonase reaction (Invitrogen). YAP1 mutant sequences were generated by gene synthesis in pDONR223 (Biocat GMBH) and subsequently cloned into pcDNA5/FRT/TO/SH/GW as described above. The sequences of YAP1 mutants oligonucleotides are reported in the Reagents and Tools table.

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Uliana F., Ciuffa R., Mishra R., Fossati A., Frommelt F., Keller S., Mehnert M., Birkeland E.S., van Drogen F., Srejic N., Peter M., Tapon N., Aebersold R, & Gstaiger M. (2023). Phosphorylation‐linked complex profiling identifies assemblies required for Hippo signal integration. Molecular Systems Biology, 19(4), e11024.

Publication 2023

LR clonase reaction

Enzymatic Oligonucleotides Proteins Proteins a Proteins n Strep Synthesis gene Vectors Yap1

Corresponding Organization :

Other organizations : ETH Zurich, Quantitative BioSciences, University of California, San Francisco, The Francis Crick Institute

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Variable analysis

independent variables

N-terminal Strep-HA-tagged bait proteins
YAP1 mutant sequences

dependent variables

Not explicitly mentioned

control variables

Gateway destination vectors (pcDNA5/FRT/TO/SH/GW)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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