In this paper, a series of polyamide derivatives (PAMs) containing morpholine groups were prepared by Ugi polymerization from dialdehyde, diacid, N-(2-aminoethyl)-morpholine and isonitrile compounds as novel multi-responsive fluorescent sensors. As non-conjugated light-emitting polymers, PAMs were endowed with unique polymerization-induced emission (PIE) performance at 450 nm by through-space conjugation (TSC) between heteroatoms and heterocycles. It was also found that PAMs exhibited reversible responses to the external temperature and pH values and became responsive fluorescent switches. In addition, PAMs can specifically recognize Fe3+ with a limit of detection (LOD) of 54 nM and the introduction of EDTA reversibly restores the fluorescence of the quenched PAMs-Fe3+ system. By virtue of thermosensitivity, PAMs are easily separated from the above system by changing the temperature above or below the lower critical solution temperature (LCST). It is worth noting that PIE-active PAMs with good biocompatibility can selectively accumulate in lysosomes due to the presence of morpholine groups, and its Pearson colocalization coefficient is as higher as 0.91. Furthermore, a PIE-active PAM was successfully used to track exogenous Fe3+ in lysosomes. In conclusion, these multi-functional PIE-active PAMs have higher potential applications in biomedical or environmental fields.