Figure A1.

Detail schematic of the two-component assembly using pGoldenGate-SE7 as the destination vector. (A) Synthetic promoter (SP), reporter (GFP), and the pGoldenGate-SE7 destination plasmid containing the lacZα gene are put in the same reaction tube along with BsaI and the T4 DNA ligase. The recognition site for BsaI (5′-GGTCTC-3′) is shown in bold. The underlined segments are XhoI and HindIII sites flanking the lacZα gene in the destination vector. (B) Shows the partially single-stranded DNA fragment generated after the digestion with BsaI (C) Ligase is used to assemble the desired product. Note that 5′-ATGA-3′ is the first four bps of the GFP coding DNA sequence and therefore this assembly is scar-less.