Escherichia coli is a cause of foodborne illness, and the emergence of antibiotic-resistant strains poses significant risks to food safety, public health, and hospital infection control. With traditional antibiotics losing efficacy, new antimicrobial strategies are needed. This study explores colicin-based biocontrol as a potential solution. We engineered a plasmid that enables E. coli K-12 to produce Colicin V, a toxin targeting other antibiotic-resistant E. coli strains that carry a colicin resistance gene, thereby preventing it from targeting itself. The plasmid also codes for Kanamycin resistance. To assess its efficacy, we will conduct LB agar plate assays under three conditions: (1) Transformed E. coli with just LB agar to assess E. coli growth; (2) transformed E. coli with LB Agar and Kanamycin to assess plasmid uptake and resistance to Kanamycin; (3) non-transformed E. coli with LB Agar and Kanamycin to assess the effectiveness of the antibiotic;(4) transformed E. coli with Kanamycin and IPTG to assess bacterial growth in the presence of colicins. By analyzing E. coli plasmid uptake, growth, and survival, we can add to existing research about colicin production in E. coli, and determine this specific E. coli strain’s capabilities of colicin production.