To collect target gene sequences, we downloaded confirmed the full cDNA sequences of VGSC, AChE, RyR, and nAChR from InsectBase (Yin et al., 2016). Next, these confirmed target gene sequences were used as queries to BLASTP against the NCBI GenBank for each target each species. The first search step obtained target sequences for species of most orders. Then, we selected the obtained target sequences from species with annotated genome as the secondary queries to search against other species within the same order. These two step searches yielded most sequences of four targets in the tested species. For species still without target sequences, we used the target sequences from the closely related species as the tertiary queries to search against the genome of this species.
To collect detoxification gene sequences of different species as comprehensively as possible, genome official gene set (OGS) files for the species were downloaded. Then, we selected all the sequences annotated as “cytochrome P450” or “glutathione S-transferase” or “carboxyl/cholinesterase”. For some important insect species without published genome OGSs, the detoxification gene sequences were obtained by directly searching against NCBI nucleotide database with terms: (((cytochrome P450) OR glutathione S-transferase) OR carboxyl/cholinesterase) AND “species name” [Organism].