The genomic and cDNA sequences of the MSCP14.6 gene section, which encodes a Manduca sexta cuticle protein, have been determined. The genomic sequence presented here includes 1428 N of 5' flanking DNA, 1587 N of transcribed DNA, and 465 N of 3' flanking DNA. The intron/exon boundaries mapped by comparison of these sequences were confirmed by primer extension and SI nuclease mapping. Prior work showed that the RNA produced by this gene is regulated both temporally and spatially during larval, pupal, and adult development, and also showed that both 20-hydroxyecdysone (20E) and juvenile hormone (JH) are important in controlling the amount of RNA present in the epidermis (Riddiford et al., 1986). In situ hybridizations show that MSCP14.6 is expressed throughout the epidermis in larvae and is spatially restricted in pharate pupal and pharate adult stages. The results presented here delimiting the transcription unit and 5' flanking DNA of the MSCP14.6 gene will facilitate further research to determine the DNA elements necessary for regulation of gene expression.