Accessibility is one of the most important factors in a location choice decision of a firm. However, new and relocated establishments can value accessibility differently. Research papers differences of accessibility preferences between creations and relocations are scarce. In this paper, we examine these differences in an intraurban setting, the Lyon urban area. We rely on discrete choice models and data for more than 43,000 creations and 11,000 relocations during 2005–2011 from eight economic sectors. Results demonstrate that the effect of accessibility differs between creations and relocations of the same economic sector. This difference depends on the type of the economic activity of the sector.