Objective:To clone the promoter sequences of stimulator of interferon gene (STING) and evaluate its activity, and to preliminarily investigate the transcriptional regulatory mechanisms. Methods: Promoter region was predicted by bioinformatics methods, and the 1 005 bp (-927~+77) fragment of 5′ upstream sequences of STING gene was amplified by PCR, and then cloned to pGL3-basic vector to construct the luciferase report gene recombinant plasmid. Three promoter fragments with different length were obtained by walking deletion and cloned into pGL3-basic vector. The vector expression activities were determined by transfection of the mouse NIH3T3 cells with the recombinant plasmids of STING gene promoter. Bioinformatics methods were performed to predict the potential transcriptional factor binding sequences. Resluts: The luciferase reporter gene recombinant vectors of mouse STING promoter were successfully constructed. Compared with the pGL3-basic plasmid, the relative luciferase activities of recombinant vectors of STING promoter were much higher (P < 0.05). In addition, the binding sequences of GATA, IK2, MZF1, SP1/SP3, and STAT may be included in the promoter region (-177~-48) of STING gene,which were predicted by the bioinformatics method. Conclusion: The luciferase report gene recombinant plasmids of STING gene promotor were constructed successfully and had strong transcriptional activity in NIH3T3 cells. By the activity comparison, it is speculated that the core promoter region of mouse STING is located in the -177~+77 region, which may contain a number of potential transcription factor binding sequences.