Pseudoalteromonas piscicida strain DE2-B is a naturally occurring, Gram-negative, marine Gammaproteobacterium that was isolated from seawater from the Delaware coast (USA). It produces antimicrobial compounds that have a broad spectrum of inhibitory activities against human and fish pathogens including vibrios. Pseudoalteromonas spp. have been used as probiotics to reduce pathogens in aquaculture and has been shown to reduce biofilm formation. In this study, we fully sequenced and annotated P. piscicida strain DE2-B to further elucidate the genome and to facilitate research on compounds that are inhibitory toward other Gram-negative as well as Gram-positive bacteria.