The ZNF313 gene has the highest transcription level in fertile male testes and may be related to human spermatogenesis. The deletion-mutated plasmids of ZNF313 promoter were constructed and transfected into HEK293 cells. The result showed that the fragment from nt −157 to +8 has a basal transcriptional activity. A functional Sp1 binding site was identified by site-directed mutation test and mithramycin A treatment. A 447-bp based at +233 to −213 exhibits a characteristic CpG island, which overlaps with the promoter region. Our work suggests that ZNF313 is controlled at the transcriptional level, and a common mechanism controlling the basal transcription of ZNF313 gene exists.