Cloning of dTAG-tagged KRAS and LACZ

Cloning of pLEX_305-N-dTAG-KRAS^G12V was previously described.^{26 (link)} pLEX_305-C-dTAG-LACZ was generated by cloning LACZ (Addgene, #25893) into pLEX_305-C-dTAG using gateway recombination cloning technology (Invitrogen) as previously described.^{26 (link)} In addition to an N- or C-terminal FKBP12^F36V tag, respectively, these plasmids contain tandem HA tags for monitoring of protein expression and a puromycin selectable marker.

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Ferguson F.M., Nabet B., Raghavan S., Liu Y., Leggett A.L., Kuljanin M., Kalekar R.L., Yang A., He S., Wang J., Ng R.W., Sulahian R., Li L., Poulin E.J., Huang L., Koren J., Dieguez-Martinez N., Espinosa S., Zeng Z., Corona C.R., Vasta J.D., Ohi R., Sim T., Kim N.D., Harshbarger W., Lizcano J.M., Robers M.B., Muthaswamy S., Lin C.Y., Look A.T., Haigis K.M., Mancias J.D., Wolpin B.M., Aguirre A.J., Hahn W.C., Westover K.D, & Gray N.S. (2020). Discovery of a selective inhibitor of Doublecortin Like Kinase 1. Nature chemical biology, 16(6), 635-643.

Publication 2020

gateway recombination cloning technology

Lacz Plasmids Protein Puromycin Recombination

Corresponding Organization :

Other organizations : Dana-Farber Cancer Institute, The University of Texas Southwestern Medical Center, Harvard University, Broad Institute, Beth Israel Deaconess Medical Center, Baylor College of Medicine, Universitat Autònoma de Barcelona, Promega (United States), University of Michigan–Ann Arbor, Korea University, Korea Institute of Science and Technology

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Variable analysis

independent variables

Cloning of pLEX_305-N-dTAG-KRAS^G12V
Cloning of pLEX_305-C-dTAG-LACZ

dependent variables

Monitoring of protein expression

control variables

Gateway recombination cloning technology (Invitrogen)
FKBP12^F36V tag
Tandem HA tags
Puromycin selectable marker

Annotations

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