“…Furthermore, the precise boundaries of protein coding genes (PCGs), tRNA, and rRNA were determined by aligning the individual genes with MUSCLE v3.8.1551 ( Edgar, 2004 ) as implemented in Seaview v4.7 ( Gouy, Guindon & Gascuel, 2010 ). We used the annotated gene sequences of other beetle mitogenomes available in Genbank as references to assess the 5′ and 3′ ends of PCGs and rRNAs: Tribolium castaneum , NC_003081 ( Friedrich & Muqim, 2003 ), Eucryptorhynchus brandti , NC_025945 ( Nan, Wei & He, 2016 ), Eucryptorrhynchus chinensis NC_026719 ( Liu et al, 2016 ), Anoplophora glabripennis , DQ768215 (unpublished), Sphenophorus sp., GU176342 ( Song et al, 2010 ), Naupactus xanthographus GU176345 ( Song et al, 2010 ), Rhynchophorus ferrugineus , KT428893 ( Zhang et al, 2017 ), Sitophilus oryzae , KX373615 ( Ojo et al, 2016 ). The annotation of each tRNA gene was validated as accurate if its secondary structure matched the canonical cloverleaf structure of tRNAs in metazoans.…”