Molecular Cloning of oatA and cysB Genes

All plasmids used in this work are listed in Table 2. Primers were synthesized by Integrated DNA Technologies, Inc. (IDT [Coralville, IA, United States]) and are listed in Table 3. Genes were amplified from S. enterica genomic DNA using Pfu Ultra II fusion DNA polymerase (Stratagene) per manufactures instructions. A high efficiency cloning method described elsewhere (Galloway et al., 2013 (link); VanDrisse and Escalante-Semerena, 2016 (link)) was used to clone oatA and cysB genes into pCV3 (complementation vector, L-(+)-arabinose inducible) and pTEV16 or pTEV18 (overexpression vectors, IPTG inducible). Restriction enzyme BspQI was purchased from New England Biolabs. The resulting complementation vector was referred to as pOatA6. The resulting overexpression vectors were referred to as pOatA4 and pCysB2 (full length CysB).