AbstractWe have determined the sequence of a 10275 bp DNA segment of Yarrowia lipolytica located on chromosome VI. The sequence contains six complete open reading frames (ORFs) longer than 100 amino acids and two more partial ORFs at both ends. Two of the ORFs encode for the well‐characterized genes YlURA5 (orotate phosphoribosyltransferase) and YlSEC65 (encoding a subunit of the signal recognition particle). These two genes show an identical organization—located on opposite strands and in opposite orientations—in four yeast species: Saccharomyces cerevisiae, Kluyveromyces lactis, Candida albicans and Y. lipolytica. One ORF and the two partial ORFs code for putative proteins showing significant homology with proteins from other organisms. YlVI‐108w (partial) and YlVI‐103w show 39% and 54% identity, respectively, with YDR430c and YHR088w from S. cerevisiae. YlVI‐102c (partial) shows significant homology with a matrix protein, lustrin A from Haliotis rufescens, and with the PGRS subfamily (Gly‐rich proteins) of Mycobacterium tuberculosis. The three remaining ORFs show weak or non‐significant homology with previously sequenced genes. The nucleotide sequence has been submitted to the EMBL database under Accession No. AI006754. Copyright © 2001 John Wiley & Sons, Ltd.