Figure 1.

Construction of the integration plasmid, pJL1147, for allelic replacement of bca as described. This construct was prepared to serve as a shuttle/suicide vector for replacing the bca structural gene with a selectable kanamycin marker. pJL1147 contains pVE6007, a pUC19 origin of replication, a kanamycin resistance cassette, and the N- and C-terminal flanking regions from bca. oriTs, Ts origin of replication; oripUC, pUC19 origin of replication; Cm, chloramphenicol-resistance marker; Km, kanamycin-resistance marker; N-flanking, PCR fragment upstream from the bca structural gene; C-flanking, PCR fragment downstream from the bca structural gene. The figure is not drawn to scale.