Diagram of the original TAR cloning protocol (Larionov et al., 1997; Kouprina, & Larrionov, 2008). In Step 1, genomic DNA and TAR vector are prepared. In Step 2, genomic DNA containing a region of interest (in red) is co-transformed with the linearized TAR vector into yeast spheroplasts. TAR vector contains a YAC (yeast artificial chromosome) cassette for proper selection, segregation, and propagation in S. cerevisiae and two targeting sequences (hook 1 and hook 2 in green) homologous to the 3’ and 5’ ends of the target region. In Steps 3 and 4, and after co-transformation, recombination between the hooks and the target sequences leads to the rescue of a region of interest as a circular YAC molecule.