Antibiotic resistance is problematic for the modern world, and new antibiotic targets are exceedingly necessary. trans-Translation provides a novel target for antibiotic development as it is a ribosome-rescue pathway present in bacteria and absent in mammals. KKL-1005, and inhibitor of Mycobacterium tuberculosis, was previously shown to inhibit trans-translation. This project aims to identify the molecular target of this compound in E. coli using chemical biology techniques and confirms that the compound is not inhibiting translation.