[Objective] This study aimed to construct a genetically engineered Saccharomyces cerevisiae strain for inositol production by overexpressing its INO1 gene that encodes inositol-1-phosphate synthase. [Methods] The integrated multi-copy expression vector pURIH was constructed via an rDNA-mediated method and transformed into S. cerevisiae Y01 strain. The expression levels of INO1 in genetically engineered strains were analyzed through quantitative RT-PCR. was detected through HPLC. [Results] Two genetically engineered strains, namely, YI2-1 and YI2-2, were constructed. These strains could highly express INO1. YI2-1 produced 16.235 times more inositol than Y01 did. The KanMX resistance gene was knocked out in the YI2-1ΔKP strain, which efficiently produced up to 627 mg/L inositol. [Conclusion] pURIH could be applied to overexpress homologous INO1 in S. cerevisiae. The amount of inositol produced by the engineered strain was sufficient and thus could be employed in engineering applications.