Efficient generation and rapid isolation via stoplight recombination of Herpes simplex viruses expressing model antigenic and immunological epitopes

► A system for producing a recombinant HSV-1 expressing exogenous epitopes was created. ► Traffic-light inspired fluorescent markers gauged the purity of recombinant viruses. ► Adaptability was demonstrated by generating recombinants for 3 immunologic epitopes. ► Recombinants were rapidly identified and isolated to purity via stoplight indicators. ► Presentation of a CD8 epitope was quantified using a T cell hybridoma reporter cell.