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Sonic hedgehog (SHH), Indian hedgehog (IHH) and Desert hedgehog (DHH) are Hedgehog family ligands, which play key roles in embryogenesis and carcinogenesis, especially in pancreatic cancer and gastric cancer. Drosophila Costal-2 (Cos2) is a Hedgehog signaling regulator, interacting with Smoothened (Smo), Cubitus interruptus (Ci), Fused (Fu) and microtubule. KIF27 is one of the mammalian ortholoas for Drosophila Cos2. KIF7 gene is the paralog of KIF27 gene; however, complete coding sequence (CDS) of KIF7 remains elusive. Here we characterized human KIF7 gene by using bioinformatics. Human LOC374654 cDNA (NM_198525.1) was a 5’-truncated partial KIF7 cDNA, corresponding to the nucleotide position 1418-4538 of KIF7 complete CDS. KIF7 gene, consisting of 19 exons, was found to encode a 1343-amino-acid KIF7 protein. Human KIF7 showed 43.6% total-amino-acid identity with human KIF27. Kinesin motor catalytic (KISc) domain (codon 15-347 of KIF7) and novel KIF7-KIF27 homologous (KIF727H) domain (codon 699-1196 of KIF7) were well conserved between KIF7 and KIF27. Three coiled-coil regions were located within the KIF727H domain. KIF7 and KIF27 were N-type Kinesins with the KIF727H domain. KIF7 mRNA was expressed in embryonic stem (ES) cells, melanotic melanoma and Jurkat T cells, while KIF27 mRNA was expressed in testis, pancreatic islet, germ cell tumors and Jurkat T cells. This is the first report on the characterization of the KIF7 gene as well as on the identification of KIF727H domain.