[Background] Natamycin is a natural, broad-spectrum and efficient polyene macrolide antifungal antibiotic. Streptomyces gilvosporeus is an important natamycin-producing bacterium. However, the genome sequence analysis of S. gilvosporeus has not been reported until now, limiting studies on the biosynthesis and regulation of natamycin and other secondary metabolites in S. gilvosporeus. [Objective] The genomic sequence information of the S. gilvosporeus F607 (a natamycin high-producing strain) was analyzed to explore the genetic resources of secondary metabolite genes and lay a foundation for further study on the mechanisms of high producing and regulation of natamycin biosynthesis. [Methods] The genome sequence of F607 was analyzed with softwares to predict genes, to annotate function of genes, to analyze phylogenetic tree and colinear analysis, and to predict the secondary metabolite synthetic gene cluster; The differences of natamycin biosynthetic gene clusters in different strains were analyzed and compared through annotating analysis of natamycin biosynthetic gene clusters; the biosynthetic pathway of natamycin was analyzed and predicted according to gene function. [Results] The complete genome sequence of F607 is 8 482 298 bp ((G+C)mol%, 70.95%). 5 062, 4 428, 5 063 genes were respectively predicted in COG, GO and KEGG databases. The antiSMASH software predicted that there are 29 secondary metabolite biosynthetic gene clusters in F607 genome. The homology of natamycin biosynthetic gene cluster in F607 shared 81% and 77% with that in S. natalensis and S. chattanoogensis respectively. Although there were differences among 2 regulatory genes (sngT, sgnH) and 9 unknown function genes (orf1?9), the analysis of natamycin biosynthetic gene clusters in different strains indicated that the other genes and their arrangement in natamycin biosynthetic gene cluster are highly conserved. [Conclusion] In this study, the complete genome sequence of strain F607 was first analyzed and the biosynthetic pathway of natamycin of F607 was predicted. This study provides basic data for analysis of the molecular mechanism of high-yield natamycin in S. gilvosporeus F607, and aids in the development of useful strategies for revealing the mechanism of high yield of natamycin, improving industrial strains and innovating drug discovery.