This study seeks to find the influence of replacing a portion of the asphalt–rubber binder with the bio-based material “guayule resin.” This replacement could be beneficial in terms of sustainability, economics, and environmental concerns related to the asphalt industry. Nine asphalt-rubber-guayule binders were investigated to find their rheological properties. Consecutively, the study proceeded with five selected binders being compared to the original asphalt (PG64-22). Investigations underwent whole matrices (crumb rubber modifier (CRM) residue included) and liquid phases (CRM residue extracted). Additionally, these properties were partially sought for their corresponding asphalt–rubber binders to compare and judge the contribution of the guayule resin. Likewise, a thermo-gravimetric analysis was done for the guayule resin to recognize its moisture and composition complexity. Such an analysis was also done for the as-received CRM and some extracted CRMs to determine the release and residue of rubber components. Outcomes showed that the guayule resin has the potential to compensate the performance required against the original asphalt at elevated temperatures while greatly decreasing the asphalt cement proportion. For instance, a blend of 62.5% asphalt, 12.5% CRM, and 25% guayule resin provided better performance than that of the original asphalt.