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A deletion cassette with sequence upstream and downstream of gspD was generated by overlap PCR using primers 2514 EcoRI A and 2514 B for the upstream fragment, and 2514C and 2514D for the downstream fragment (Table S4) using Pfu Ultra polymerase (Agilent, Santa Clara, CA) and the DK1622 strain of M

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Half of the trials were congruent (24/48), where the numbers shown matched the number of ciphers (e.g., 333) and half of them were incongruent (e.g., 222)