Decreased marine survival is identified as a component driver of continued declines of Atlantic Salmon Salmo salar. However, estimates of marine mortality often incorporate loss incurred during estuary migration that may be mechanistically distinct from factors affecting marine mortality. We examined movements and survival of 941 smolts (141 wild and 800 hatchery-reared fish) released in freshwater during passage through the Penobscot River estuary, Maine, from 2005 to 2013. We related trends in estuary arrival date, movement rate, and survival to fish characteristics, migratory history, and environmental conditions in the estuary. Fish that experienced the warmest thermal history arrived in the estuary 8 d earlier than those experiencing the coolest thermal history during development. Estuary arrival date was 10 d later for fish experiencing high flow than for fish experiencing low flow. Fish released furthest upstream arrived in the estuary 3 d later than those stocked further downstream but moved 0.5 km/h faster through the estuary. Temporally, movement rate and survival in the estuary both peaked in mid-May. Spatially, movement rate and survival both decreased from freshwater to the ocean. Wild smolts arrived in the estuary later than hatchery fish, but we observed no change in movement rate or survival attributable to rearing history. Fish with the highest gill Na⁺, K⁺-ATPase activity incurred 25% lower mortality through the estuary than fish with the lowest gill Na⁺, K⁺-ATPase activity. Smolt survival decreased (by up to 40%) with the increasing number of dams passed (ranging from two to nine) during freshwater migration. These results underscore the importance of physiological preparedness on performance and the delayed, indirect effects of dams on survival of Atlantic Salmon smolts during estuary migration, ultimately affecting marine survival estimates.