Generation of Inducible Ets-1 Lentiviral Vector

The inducible Ets-1 lentiviral vector was created using gateway cloning as described earlier [48 (link)]. Briefly, human Ets-1 cDNA was amplified from pDONR223_ETS1_WT (Addgene # 82118) using primers attB1-hEts-1: GGGGACAAGTTTGTACAAAAAAGCAGGCTACCATGAAGGCGGCCGTCGATCTCAAGCCGACTCTCAC and attB2-Myc-hEts-1 GGGGACCACTTTGTACAAGAAAGCTGGGTCTATTACAGATCCTCTTCTGAGATGAGTTTTTGTTCCTCGTCGGCATCTGGCTTGACGTCCAGCATGGCGTGCAGCTCC and cloned into pDONR221 vector (Life Technologies, USA, # 12536017) using BP Clonase (Life technologies, USA # 11789100), to make the entry clone. The LR recombination reaction was performed with the entry clone using LR Clonase (Life technologies, USA, # 11791020) to generate the doxycycline-inducible Ets-1 lentiviral vector in pLIX_403 backbone (Addgene # 41395).