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. 2005 Nov;71(11):7196–7202. doi: 10.1128/AEM.71.11.7196-7202.2005

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FIG. 1. — Plasmid used in this study. Plasmid pD70 is an endogenous plasmid of M. hemolytica. Plasmid pD70Kan^R was constructed by inserting the kanamycin resistance cassette into the modified HindIII site of pD70. Plasmid pD70Kan^R was mutagenized with hydroxylamine, and the TS (Ts) plasmid pGA301 was selected after screening in M. hemolytica. Plasmid pGA301oriKan^r was constructed by digesting pGA301 with Sau3A. The resulting 1,160-bp Sau3A fragment containing the TS origin of replication was joined to the kanamycin resistance cassette of Geneblock. Plasmid pGA301oriKanC was constructed by inserting the ColE1 origin of replication into pGA301oriKan^r.