Although microscopic self-organization of simple molecular components can generate structures that exhibit the fundamental characteristics of autonomous behavior, the physicochemical origins of such autonomy remain largely unexplored. It was previously reported that gasses, including dimethyl-sulfide, were produced in a co-culture system of antibiotic crown cells and living cells in the presence of gas-exposed sphingosine-DNA-Adenosine. In this study, using gas-exposed sphingosine, the order-dependent assembly of gas-exposed sphingosine, E. coli DNA, and adenosine were used to clarify how autonomy may develop in non-living chemical systems. Among several approaches examined, adding gas-exposed sphingosine, DNA, and then adenosine produced the most distinct and reproducible behavior. These findings revealed an order-dependent, self organizing pathway of interactions without genetic replication or metabolism. The method provides a promising model for the emergence of autonomous behavior. Gas-exposed sphingosine was referred to as G-Sph, and crown cell seeds prepared with G-Sph were named G-crown cell seeds. The resulting cells were named G-Sph- coli crown cells, reflecting the DNA used in the preparation of crown cell seeds.