While urban accessibility of museums plays a crucial role in the growth of a cultural city, in reality, an uneven distribution of museums exists in cities. In particular, museums are concentrated in certain regions or located in a place that is different from the cognitive experience of local residents. To solve this issue, this study quantified the urban space of Seoul, which has entered into the status of a cultural city since the 1990s, as its target city by using space syntax. Further, a suitability analysis was conducted by extracting the museums’ topological accessibility in the city structure as well as the accessibility by the travel angle and limit distance setting. The results showed that the physical locations of museums considering the minimum walking distance set in this study were somewhat separated or isolated from primary spaces where people travel. This indicates that determining museum locations by referring to the major travel routes throughout the city is a significant basis for securing physical accessibility. This study is meaningful as it establishes reference materials for determining museum locations in Seoul and will help form physical clusters of museums adjacent to each other.