Three single-copy sequences from the malaria vectorAnopheles gambiaeGiles are described. For each clone, a restriction enzyme site map, a genomic Southern analysis, and localization to polytene chromosomes are presented. Two clones revealed restriction site fragment polymorphisms, whereas another showed no variation within the strains tested. A portion of 1 clone was sequenced, but showed no significant homology to other sequences in GenBank. The sequence is available in GenBank as accession number L11314. The clones mark chromosome arms 2L, 2R, and 3L.