Sequential rounds of targeting inserted the selection cassettes and loxP sites proximal to Herc2 and distal to Mkrn3. A double-targeted clone in which the targeting had occurred on the homologous chromosomes (trans) was transiently transfected with a Cre recombinase in ES cells to induce recombination between the loxP sites on opposite chromosomes, which generated clones with a balanced duplication and deletion of 6.3 Mb region between Herc2 and Mkrn3. Only duplicated allele can be recovered after meiosis and mating. (J:151648)