Figure 2. Donor plasmid design for the generation of GR^dim KI rat.

Upper DNA indicates the wild type sequence of the exon 3 of the rat Nr3c1 GR. In blue the residue A476. TAL 3 and TAL 6 binding sites are indicated. DP, donor plasmid sequence. Donor plasmid was synthesized with 500 pb homology arms on both ends. The pA476T point mutation is highlighted in blue. Silent mutations of the DP are shown in red bold letters and are located in the overlap of TAL 3 and Tal 6 binding sites. HaeIII site is highlighted in yellow. AluI site is not shown in this figure.