MYB transcription factors play a crucial role in regulating plant development and adaptation. Brassica napus, a major oil crop, often faces challenges such as lodging and diseases. In this study, using the genome of Brassica napus as a template, we cloned a 2000 bp promoter sequence upstream of the start codon of the pBnMYB69-1 gene. Prediction and analysis of cis-regulatory elements revealed that the promoter contains essential core regulatory elements of eukaryotic promoters, such as the TATA box and CAAT box. Additionally, it includes various functional elements, such as light-responsive elements (ATCT-motif, Box 4, TCT-motif), anaerobic induction elements (ARE), methyl jasmonate (MeJA)-responsive elements (CGTCA-motif, TGACG-motif), and gibberellin-responsive elements (P-box). These findings provide a solid foundation for the application of this promoter in transgenic improvement of crop quality and the artificial creation of germplasm resources in Brassica napus.