De novo DNA synthesis plays crucial roles in life science. Enzymatic oligonucleotide synthesis (EOS) has attracted interest due to longer synthesized chains, simple procedure, cost-effectiveness, and environmental friendliness. However, unlike chemical synthesis dominated by small molecule, the EOS relies on enzyme reacting with primers. It remains challenging due to restricted accessibility caused by the anisotropy of initiator primers and the spatial hindrance of enzymes. Herein, this study developes a nanoscopic interface functioned with 3D DNA framework to achieve efficient EOS. The highly...