Pseudomonas aeruginosa strain Boston 41501 was isolated from blood culture. This whole-genome sequenced bacterial strain has applications in media and susceptibility testing, evaluation of Mueller-Hinton Agar, and as a quality control strain for Abbott, API, Autobac, BBL, bioMerieux Vitek, Micro-Media, MicroScan™, Roche Diagnostics, and Sensititre products.