Biofilm formation caused by the fouling of microorganisms is one of the major problems in biomedical devices, food industry, and marine transportation. Since the removal of adherent biofilm is not a trivial task, it is of paramount importance to contain the formation of the anti-biofilm film. Herein, a polysulfide-based anti-biofilm coating (PAC) equipped with full transparency, non-toxicity, and environmental stability was developed via a simple vaporphase synthesis. The polymer coating consists of polysulfide chain grafted onto poly (1,3,5,7-tetramethyl-1,3,5,7-tetravinyl cyclotetrasiloxane) (pV4D4) layer via thiol-ene click reaction, which was accomplished via a sequential deposition of each polymer followed by UV irradiation. The pV4D4 served as an adhesion promoter layer that substantially enhanced the interfacial adhesion between polysulfide layer and various substrate materials. The polysulfide layer exhibits a long-lasting anti-biofilm performance against pathogenic bacteria, such as Escherichia coli: O157 and Staphylococcus aureus. The excellent anti-biofilm property is attributed to slippery surface derived from the non-adherent, dynamic characteristics of the polysulfide (-S-S-) chain. The anti-biofilm coating indeed shows outstanding durability and robustness when exposed to extreme pH, organic solvents, and mechanical stresses. The fully transparent, robust coating developed in this study is a promising candidate material for a broad range of anti-biofilm applications.