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A deletion cassette with sequence upstream and downstream of gspD was generated by overlap PCR using primers Δ2514 EcoRI A and Δ2514 B for the upstream fragment, and Δ2514C and Δ2514D for the downstream fragment (Table S4) using Pfu Ultra polymerase (Agilent, Santa Clara, CA) and the DK1622 strain of M
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