Functional DNA molecules are useful components in nanotechnology and synthetic biology. To expand the toolkit of functional DNA parts, in this study we used artificial evolution to identify a glowing deoxyribozyme called Supernova . This deoxyribozyme transfers a phosphate from the 1,2-dioxetane substrate CDP-Star to its 5’ hydroxyl group, which triggers a chemiluminescent reaction and a flash of blue light. Comparative sequence analysis of 135,000 variants of Supernova obtained using in vitro selection and high-throughput sequencing indicated that the catalytic core of the deoxyribozyme is made up of 38 conserved nucleotides and 46 total positions. It also revealed the unusual secondary structure of Supernova, which consists of a purine motif triple helix and two purine-rich elements. An engineered version of Supernova can be programmed to only produce light in the presence of an oligonucleotide complementary to its 3’ end, demonstrating that catalytic activity can be coupled to ligand binding. We anticipate that Supernova will be useful in a wide variety of applications, including as a signaling component in allosterically regulated sensors and in logic gates of molecular computers.