Figure 1

Assembly of bi-cistronic operons. (A) Basic parts used in the assembly. ORFs and tags were synthesized as either N- or C-terminal sets. N-terminal sets contain RBSs as a component of the part, while C-terminal sets contain stop codons at the end of the part. (B) Sample combinatorial architectures for a set of constructs containing two ORFs (Med7 in the first position and Med21 in the second position) tagged at the N- or C-terminus with different tags (VSV-tag for Med7 and E-tag for Med21). Every operon is composed of 8 basic parts. For this arrangement, 4 different construct architectures are possible: in the NN configuration both ORFs are tagged at the N-terminus; in the NC configuration the first ORFs is tagged at the N-terminus and the second ORF is tagged at the C-terminus; in the CN configuration the first ORFs is tagged at the C-terminus and the second ORF is tagged at the N-terminus; in the CC configuration both ORFs are tagged at the C-terminus.