The production of lasso peptides has been considerably expanded through the development of heterologous expression and cell-free biosynthesis approaches. However, an adaptable chemical synthesis platform would provide further access to this scaffold and unnatural variants. A central limitation in the development of a chemical synthesis platform is the requirement that the lasso precursor be pre-folded prior to isopeptide bond formation; otherwise, the non-threaded conformation (herein referred to as a tadpole) would be obtained. Indeed previous direct cyclization attempts to access lasso peptides have exclusively given the non-threaded tadpole isomers. While biological systems stabilize the pre-lasso fold through an enzyme-substrate complex, the outstanding question explored in this work is whether the lasso precursor can also be sufficiently stabilized through the intramolecular forces that dictate peptide and protein folding.