Herein, we have successfully prepared sequence-controlled polymers with versatile succimide motifs by one-shot reversible addition-fragmentation chain-transfer (RAFT) polymerization of styrene and furan-protected maleimide. Succimide motifs at desired positions along the polymer chain were highly reactive for post-modifications in both nucleophilic substitution and Mitsunobu condensation reactions.H-1 NMR, FT-IR, MALDI-TOF mass spectrum and SEC measurements were used to demonstrate the efficiencies of the nucleophilic substitution reaction and Mitsunobu condensation reaction. Then, we used methoxy terminated polyethylene glycols (mPEG, M-n = 400 Da) as side chains to prepare sequence-controlled amphiphilic brush polymers by performing Mitsunobu condensation reaction. There was no doubt that the reactions were extremely efficient. Therefore, we could design varied side chains to prepare sequence-controlled brush polymers, which would advance the structure-property relationship and potential applications of sequence-controlled polymers in research.