[Objectives]  The whitefly, Bemisia tabaci, has developed resistance to a wide range of insecticides, including organophosphates, pyrethroids, insect growth regulators, and especially neonicotinoids. To understand the molecular mechanisms underlying neonicotinoid resistance, the genomic DNA fragment of the cytochrome P450 gene cyp6cm1 and its 5′ flanking sequence were cloned in Q-biotype B. tabaci fromYangzhou. [Methods]  PCR and genome walking were used to isolate the genomic DNA fragment of the cyp6cm1 gene and its flanking sequence. [Results]  The genomic fragment of the cyp6cm1 gene contained the last 63 bp of the first exon of the cyp6cm1 gene together with the first 826-829 bp of the adjacent intron. Three single-nucleotide polymorphisms (SNPs) associated with resistance to neonicotinoid insecticides were identified by multiple sequences alignment, these were located at positions 195, 230 and 242 of the intron, respectively. In addition, the 962 bp 5′ flanking sequence of cyp6cm1 gene of Q-biotype B. tabaci was cloned by genome walking. NNPP online analysis software predicted the transcription initiation site as the nucleotide A, which is located 57 bp upstream of the initiation codon. ConSite software predicted several transcription factor binding sites in this region, including XRE-AhR, CREB, Oct-1 and Broad-complex-4.