Plasmid pSG1154-apoaequorin was constructed using GeneArt Seamless Cloning and Assembly Kit (Life Technologies). Briefly, the encoding gene for the apoaequorin fragment (40) was chemically synthesized by GenScript, followed by assembling to the pSG1154 vector. The construction was confirmed by sequencing and then integrated into the wild-type B. subtilis amyE locus of chromosome using the standard transformation method (41).

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