A loxJTZ17 site was inserted at the 5' end of a cassette containing the NLS-cre-polyA sequence (nuclear localization signal-cre recombinase-polyadenylation signal). A PGK-puromycin selection cassett and a lox2272 site was inserted at the 3' end of the targeting construct. Targeted recombination in ES cells already containing Spink3tm1Kymm inserted the cre cassette upstream of exon 1 of Spink3, replacing the neo marker. Chimeras were produced from correctly targeted ES cell clones and germline transmission was established by crosses to C57BL6/J animals. (J:205182)