The SiMYB44 gene of MYB family in foxtail millet, which responds to various abiotic stresses, was selected through transcriptome sequencing screening in the early stage. Bioinformatics prediction, subcellular localization, yeast transcriptional activation activity analysis, and downstream element binding experiments were employed to investigate the basic characteristics of this gene. The bioinformatics prediction results indicated that SiMYB44 encodes a protein consisting of 388 amino acids, with a molecular weight of 41.43 kDa and an isoelectric point pI of4.96. Phylogenetic analysis showed that SiMYB44 was relatively close to OsMYB44, but SiMYB44 was less identical to homologs other than proteins, which suggested that SiMYB44 markedly differs from other members. Subcellular localization experiments confirmed that the SiMYB44 protein is expressed in the nucleus. The full-length gene has transcriptional activation activity and can specifically bind to cis-acting elements containing MBS and AC core sequences in the promoters of genes, suggesting that SiMYB44 regulates the transcription of certain genes by recognizing these sequences. These results can provide important genetic resources and lay a theoretical foundation for transgenic breeding of foxtail millet.