Figure 1

Scheme of the two independent integration systems used. A: FRT docking site is used as in the Flp-In T-Rex™ system (Invitrogen). The FRT sequence is placed downstream and in frame of the translational initiation codon ATG from the lacZ-zeocin fusion protein. After FLP mediated recombination with the FRT sequence of the integration vector, lacZ-zeocin expression is turned off and hygromycin resistance is activated. The GOI (gene-of-interest) is placed downstream of the hygromycin resistance cassette and controlled by the tetracyclin inducible CMV promoter containing the two tetracycline operators (2xtetO). B: attP docking site for ΦC31 integrase mediated integration. The attP sequence is placed downstream and in frame of the translational initiation codon ATG from the ECFP-Neo fusion protein. After ΦC31 mediated recombination with the attB sequence of the integration vector, ECFP-Neo expression is turned off and puromycin resistance is activated. The GOI is placed downstream of the puromycin resistance cassette and controlled by the CMV promoter. The N-terminal destabilizing domain (DD) is linked to the GOI to allow regulation by Shld1.