The microwave spectrum of 3-butyne-1-thiol has been studied by means of Stark-modulation microwave spectroscopy and quantum-chemical calculations employing the B3LYP/6-311++G(3df,2pd), MP2/aug-cc-pVTZ, MP2/6-311++G(3df,2pd), and G3 methods. Rotational transitions attributable to two conformers of this molecule were assigned. One of these conformers possesses an antiperiplanar arrangement of the atoms S-C1-C2-C3, while the other is synclinal and stabilized by the formation of an intramolecular hydrogen bond between the H-atom of the thiol group and the pi-electrons of the C[triple bond]C triple bond. The energy difference between these conformers was estimated to be 1.7(4) kJ mol(-1) by relative intensity measurements, with the hydrogen-bonded conformer being lower in energy. The spectra of five vibrationally excited states of the synclinal conformer were observed, and an assignment of these states to particular vibrational modes was made with the aid of a density functional theory (DFT) calculation of the vibrational frequencies at the B3LYP/6-311++G(3df,2pd) level of theory.