The development of homogeneous proteins with novel reactivity represents a cutting-edge frontier at the interface of chemistry and biology. The development of advanced protein analogs requires versatile strategies to edit specific residues in a precise and controlled manner; 35 however, incorporating novel transformations into the native sequence of proteins is often challenging through conventional biological methods. Here, we integrated structure-based design, peptide synthesis, and chemoselective ligation to develop novel abiotic TFs with refined activity. We chemically synthesized a library of abiotic TF analogs derived from Max TF. Through comprehensive biochemical and biophysical analyses, we found that the mutation site plays a critical role in the DNA binding activity of TFs. Importantly, systematic DNA binding and cellular studies led to the discovery of μMax20, an analog bearing two mutations (Lys31/57 to hArg), which exhibited potent DNA binding to the core E-box site. Moreover, we found that the stapled advanced analog, 2s-μMax20, is amenable to cellular delivery at submicromolar concentrations in HeLa cells. The newly developed μMax20 analogs exhibited enhanced cell permeability and retained potent DNA binding activity to the canonical E-box sequence, outperforming native Max analogs. Although TFs have previously been demonstrated to be cell permeable, 37 likely due to their basic DNA-binding domains, the designed μMax20 analogs exhibited exceptional cell penetration at low concentrations, outcompeting natural TFs. While our preliminary results indicated that the penetration of 2s-μMax20 analog might have a dual mechanism of internalization involving both endocytosis and direct translocation, further mechanistic analysis is required. We believe that incorporating non-canonical transformations can be used to identify novel reactivities where a protein can tolerate non-proteinogenic amino acids without significantly affecting its folding and function, enabling the engineering of abiotic proteins with novel properties. Finally, recent reports have demonstrated the potential of synthetic TFs to manipulate oncogene expression. We anticipate that the potent DNA binding activity and novel cell permeability of the developed abiotic TFs will pave the way for developing advanced analogs for therapeutic applications.