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Stromatolites have often served as a diagnostic carbonate facies for deep‐time palaeoclimatic and geobiological studies because they may form under favourable environmental conditions for microbially mediated carbonate production. 'Giant' (<5 m) stromatolites occur in the Laney Member of the Eocene Green River Formation in the Vermillion Creek section of the Sand Wash Basin (north‐west Colorado, USA). Giant stromatolite growth was hypothesized to have been promoted by both availability of large substrates as nucleation sites and physicochemical factors, including increased calcium carbonate mineral saturation states due to the generally warmer Eocene climate and a dynamic period in Lake Gosiute's hydrological balance. Depositional horizons of giant stromatolites were observed at two distinct stratigraphic levels, which demonstrated that the formation of the giant morphotype was not a unique occurrence, and provided an opportunity to examine both onset and cessation of stromatolite development. Coincident increases in carbonate clumped isotope‐derived temperatures, the carbon and oxygen isotopic compositions of lake water (δ18Ow), carbonate mineral saturation states (Ω) estimated by ooid size, and salinity estimated by ostracod assemblages demonstrated that formation of giant stromatolites was facilitated by lake level drawdown. Decreased lake levels: (i) promoted carbonate precipitation; and (ii) positioned benthic microbial communities within the photic zone. Field and petrographic analyses revealed that the giants preserved micritic laminations, 'trapped and bound' grains, and aragonite fans, interpreted as reflecting contributions from both microbially mediated and abiotic carbonate mineral precipitation. Field and microscopic sedimentological and stable isotope data indicated that giant stromatolite growth ceased as a result of subaerial exposure of mounds during lake level lows. Although microbial mediation of carbonate chemistry was seemingly important for initiation of stromatolite growth, this work demonstrated that stromatolite macromorphology was dominantly controlled by availability of large substrates, lake level and resultant solute chemistry, i.e. increased Ω. [ABSTRACT FROM AUTHOR]