Anthropogenic structures can form novel ecosystem niches. Invasive species are often particularly successful in occupying these habitats and utilize them as beachheads for further spread. The invasive round goby (Neogobius melanostomus, Pallas 1814), an inherently bottom‐dwelling fish, uses vertical harbor walls as habitat, enabling them to reach boats (i.e., potential translocation vectors). To evaluate the relevance of vertical habitat use for population dynamics and translocation, we exemplary investigated a population of round gobies in a harbor ecosystem. Specifically, we investigated differences in trophic niche characteristics, individual trophic specialization, phenotypic traits, and breeding frequency in wall versus bottom‐dwelling round gobies. Habitat‐characteristic dietary signatures indicated habitat partitioning during the breeding season. Trophic niches overlapped but were clearly distinguishable between the habitats: Walls were inhabited by 1.4 times more trophic generalists than specialists, while the bottom was inhabited by 2.1 times more trophic specialists. Breeding frequency was 24 times higher on the walls than on the bottom. After the reproductive season, we found a higher similarity in trophic ecology of gobies inhabiting the two habitats, and differences in abundance, size, and condition. These results are in line with winter migrations to deeper habitats, which are common in round gobies in lentic and marine ecosystems. Our results suggest a high potential for microgeographic adaptation to either horizontal or vertical habitat use in invasive round gobies. We demonstrated that male gobies using the walls during the breeding season are larger and heavier, suggesting that wall climbing may select for more competitive individuals. Additionally, the overall abundance of round gobies likely increases with the additional use of vertical habitat space, which may lead to higher propagule pressure. The ability to exploit anthropogenic habitats, and a higher translocation probability of competitive individuals, can contribute to the invasion success of round gobies in anthropogenically influenced aquatic systems.