A full-length cDNA encoding house dust mite allergen Der f 7 from Dermatophagoides farina (Acari: Pyroglyphidae) from China was cloned, sequenced, and successfully expressed. A reference sequence (GenBank accession AY283292) was used to design polymerase chain reaction primers. Analysis revealed eight mismatched nucleotides in five Der f 7 cDNA clones, and the projected amino acid sequence contained six incompatible residues. These results suggest that the sequence of Der f 7 may be polymorphic. Further bioinformatic analysis revealed that the mature Der f 7 allergen had a molecular mass of approximately 21.88 kDa and a theoretical isoelectric point of 4.90. Der f 7 protein secondary structure was composed of a helix (56.63%), extended strand (5.10%), and random coil (38.27%). Group 7 allergens are present in Pyroglyphidae, Acaridae, and Glycyphagidae families, and homology analysis revealed a 86% similarity between Der f 7 and Der p 7. Furthermore, a phylogenetic tree constructed of group 7 allergens from different mite species revealed that Der f 7 and Der p 7 clustered with 100% bootstrap support. Bioinformatics-driven characterization of Der f 7 allergen as conducted in this study may contribute to diagnostic and therapeutic applications for dust mite allergies.