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Recombinants were selected on LB agar containing Kan incubated at 30 °C overnight, and genes deletion in the BAC was confirmed by PCR amplification using “Conf-KO BAC cobMK Mcan-Fw” and “P1 inv” primers for the 5’ end and “P2 inv long” and “Conf-KO BAC cobMK Mcan-Rv” for the 3’ end