Rhodopsinlike opsins constitute a distinct phylogenetic group (Yokoyama 1994, Mel. Biol. Evol. 11:32-39). This RH2 group includes the green-sensitive opsins in chicken and goldfish and the blue-sensitive opsin in a nocturnal lizard gecko. In the present study, we isolated and sequenced the genomic DNA clones for the RH2 opsin gene, rh2(Ac), of the diurnal lizard Anolis carolinensis. This single-copy gene spans 18.3 kb from start to stop codons, making it the longest opsin gene known in vertebrates. Phylogenetic analysis strongly suggests that rh2(Ac) is more closely related to the chicken green opsin gene than to the gecko blue opsin gene. This gene tree differs from the organismal tree, where the two lizard species should be most closely related, implying that rh2(Ac) and the gecko blue-sensitive opsin genes have been derived from duplicate ancestral genes.