The antibacterial peptide toxin colicin V is exported from Escherichia coli cells by a signal sequence-independent, ABC export system. Export requires at least three proteins-membrane fusion protein CvaA, ABC export protein CvaB, and outer membrane protein TolC. The cvaA gene also encodes a second protein, CvaA, initiated from an in-frame translational re-start within the cvaA coding sequence. To determine whether the internally encoded CvaA protein also functions in the export pathway, the putative start codons for CvaA were mutagenized, while maintaining CvaA function. Elimination of CvaA translation caused no change in colicin V export levels, indicating that the CvaA protein is not required in the secretion pathway.