Heat shock proteins (Hsps) are an evolutionarily conserved group of molecules present in all eukaryotic and prokaryotic organisms. Hsp10 and Hsp60 were originally described as the essential mitochondrial proteins involved in protein folding. Recent studies demonstrate that Hsp10 has additional roles including immune modulation. In our study, an homologous Hsp10 (Sp-Hsp10) was identified in the mud crab Scylla paramemosain, and its genomic DNA organization was determined. The cDNA sequence of Sp-Hsp10 contains an open reading frame of 309 bp, encoding a putative protein of 102 amino acid residues with approximately 10 kDa. The Sp-Hsp10 gene is located next to the Sp-Hsp60 gene and shares a 1916-bp intergenic region. The promoter activity of the Sp-Hsp10 flanking gene was analyzed using luciferase reporter assays in transfected endothelial progenitor cells. The upregulation of Sp-Hsp10 expression was detected after exposure of hemocytes to a heat shock of 1 h at 37 °C compared with unstressed hemocytes raised at 20 °C. To our knowledge, this is the first report characterizing the genomic organization of a new Hsp10 in a crustacean.