The invention discloses a gene sequence with a molecular switch, a plasmid and an immune cell, and belongs to the technical field of molecular biology. The gene sequence comprises an excitation component, a connection region and a response component, wherein the excitation component comprises a target binding region and a transcription regulation region; the response component comprises an expression region and a promoter region matched with the transcription regulation region; the target binding region of the excitation component is used for expressing a target binding receptor; the transcription regulation region is used for expressing a transcription regulation factor; after the target binding receptor is used for being bound with a target, the transcription regulation factor is separated from the target binding receptor, is bound to the promoter region, and activates a promoter to start expression of the gene sequence in the expression region; and the connection region connects the excitation component and the response component in the same gene sequence. The preparation flow is simplified, the transfection efficiency is improved, and industrial preparation of virus particles and CAR cells is facilitated.