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The DNA fragments containing genes th , pcd, dhpr and vector pRSF 19 were amplified using primer pairs th -Gibson-1/ th -Gibson-2, pcd -Gibson-1/ pcd -Gibson-2, dhpr -Gibson-1/ dhpr -Gibson-2 and pRSF-Gibson-1/pRSF-Gibson-2, respectively, and then were assembled using Hieff Clone Plus Multi One Step Cloning Kit (YEASEN, Shanghai, China), resulting in plasmid pRSF- th-pcd - dhpr