A carbocation cyclization/rearrangement mechanism for the biosynthesis of isothapsadiene and β-isothapsenol is shown to be energetically viable on the basis of density functional theory (DFT) calculations. In addition, for both isothapsadiene and β-isothapsenol, variable-temperature NMR experiments reveal two equilibrium conformers that undergo hindered exchange. The identities of these conformers, which are related by a chair-flip, are confirmed by DFT calculations on their structures, energies, 1H and 13C chemical shifts, and interconversion pathways.