Table 2. Primers used in this study with their sequences, annealing temperature and functions.

Primer	Sequence 5′ — 3′	TA °C	Function
SacBF2	ATGAACATCAAAAAGTTTGCAAAA	58	Amplification of sacB gene in merodiploids and to confirm the elimination of suicidal plasmid backbone in V. cholerae
SacBR	TTATTTGTTAACTGTTAATTGTCC	58	Amplification of sacB gene
KanFse-2F	AGCGGCCGGCCGCTTACATGGCGATAGCTA	56	Amplification of aphA cassette in V. cholerae
KanFse-R	ATAGGCCGGCCTCAGAAGAACTCGTCAAGA	60	Amplification of aphA
RtxC F	TTATCAGAGATGGCAGCACC	66	Amplification of rtx, ΔrtxC/A and rtx::aphA genes in V. cholerae
RtxC R	CTTGTCCACCGCTGTAGCCT	66	Amplification of rtx, ΔrtxC/A and rtx::aphA genes in V. cholerae
VHAF	ATGTCTTTGCTTGCCATTGG	56	Amplification of hemA and ΔhemA genes in V. cholerae
VHAR2	GTTCAGATCGTCAAGACCTA	56	Amplification of hemA and ΔhemA

T_A annealing temperature.