Two species of Synactinernus sea anemones were found in Japanese waters. Synactinernus flavusCarlgren, 1918, the only described species of this genus, is rediscovered from off the Goto Islands a century after the original description. Synactinernus flavus was once synonymized with Isactinernus quadrilobatusCarlgren, 1918; however, we show that, based on morphological (including examination of type specimens) and molecular (using nuclear 18S rDNA) evidence, these species are completely different. The other species, Synactinernus churaumi sp. nov., was found off Ishigaki Island and Okinawa Island by a remotely operated vehicle (ROV), and had been kept for 15 years in a tank at the Okinawa Churaumi Aquarium. There are clear differences between these two species; therefore, we describe the second species and revise the diagnosis of Synactinernus.