The targeting vector expressing the FLAG tag-HA tag-T6B-YFP transcript under control of TRE and CMV minimal promoter, was generated from a modified version of the pgk-ATG-FRT plasmid, in which the region of pgk-ATG-FRT comprised between the EcoRI site and the PciI site was substituted with the rabbit beta-globin polyadenylation signal (RBG pA). The FLAG-HA-T6B-YFP DNA insert was generated by PCR using the plasmid pIRES-Neo-FLAG-HA-T6B-YFP as a template. Next, the modified pgk-ATG-FRT, was cut with NcoI, filled-in to produce blunt ends, dephosphorylated and ligated to the PCR-generated CMV-TRE-FLAG-HA-T6B-YFP DNA fragment. T6B is a short Tnrc6b-derived peptide (amino acid position 599-683) that competes with TNRC6 protein binding. (J:314127)