The complete nucleotide sequence of the mitochondrial genome of Macrotermes barneyi Light (Isoptera: Termitidae) was determined. This mitochondrial genome is 15,940 bp with 37 typical animal mitochondrial genes and an A+T-rich region. Gene positions and directions are identical to that of the pupative ancestral arrangement of insects. All protein-coding genes start with ATN codon. Seven protein-coding genes stop with termination codon TAA. Four protein-coding genes use incomplete stop codons TA and two use T. The base composition of M. barneyi mitochondrial genome is normal to most insects. All of the 22 tRNA genes, ranging from 63 to 76 bp, have a typical cloverleaf structure except for trnS2. In trnS2, D-stem pairings in the DHU (dihydrouridine) arm is absent as in many sequenced insect species. Five elements characterized by A+T-rich region of the insect mitochondrial genome are found on their conserved arrangement. However, no repeat sequence is present on the downstream of the A+T-rich region.