A targeting construct was generated with an improved cre coding region and rabbit beta-globin poly A signal sequence inserted into the 5' UTR of Cyp11a1, 14 bp upstream of the ATG codon. The construct contained a 2.8 kb Cyp11a1 fragment with the cre/poly A insertion (with a 100 bp deletion in Cyp11a1, including the ATG site). This fragment was introduced into bacterial cells for homologous recombination with a BAC containing the Cyp11a1 gene sequence. Recombined BAC DNA was purified for injection into fertilized pronuclei. One founder line was characterized by maing with two different mouse strains carrying cre-responsive reporters. (J:189958)