Yeast ino4 mutants are auxotrophic for the phospholipid precursor inositol and have pleiotropic defects in phospholipid synthesis. The mutants are unable to derepress the cytoplasmic enzyme, inositol-1-phosphate synthase and they exhibit reduced synthesis of methylated phospholipids, particularly phosphatidyl-choline. The INO4 gene is believed to encode a positive regulator involved in coordinate control of phospholipid synthesis. In the present study, we report the isolation of two clones containing the INO4 gene. The clones share a region of homology and were mapped to the INO4 locus. Southern blot analysis revealed that the cloned DNA contained both unique and repetitive DNA.