Pou5f1^tm1Grc

A BAC containing the mouse locus was manipulated through recombineering. The construct was flanked with homology arms of approximately 3.7 kbp upstream and 10.9 kbp downstream. Two distinct DNA binding sequence arrays were added at -277 bp upstream of the transcriptional start of of the gene, 12 x ZFHD1 (TAATGATGGGCG) and 5 x Gal4 (CGGAGTACTGTCCTCCGAG). A nuclear EGFP was inserted at the ATG of exon 1, exons 2-5 were deleted after insertion to avoid expression of endogenous Pou5f1 from the reporter allele. A floxed neomycin resistance cassette was inserted behind the EGFP reporter for positive selection and thymidine kinase was inserted behind the homology arms for negative selection. (J:199709, J:199710)