E coli 5 alpha chemically competent cells

mRNA from Arabidopsis thaliana 7-day old seedlings was isolated using the Spectrum Plant Total RNA Kit (Sigma-Aldrich) according to the manufacturer’s instructions. cDNA was generated using Super Script IV First Strand Synthesis System (Invitrogen). Gene sequences for HMGR, DXS, and RuBisCo were amplified by PCR using Q5® High-Fidelity DNA Polymerase and gene specific primers form Integrated DNA Technologies (Supplementary Table 4)^{51 ,52} . All other gene sequences were synthesized by Gen9, Inc and PCR amplified as described above (Supplementary Table 5). Gibson assembly was used to insert PCR amplicons into AgeI and XhoI (New England Biolabs) linearized pEAQ-HT vector⁵³ . All synthesized genes were designed with 5’ pEAQ-HT vector overlaps necessary for Gibson assembly. Constructs were transformed into either E. coli Top10 chemically competent cells (Invitrogen) or E. coli 5-alpha chemically competent cells (New England Biolabs). Plasmid DNA was isolated using the QIAprep Spin Miniprep Kit (Qiagen). Sequence confirmation was carried using Sanger DNA sequencing (Elim Biopharm).