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Figure S1. cDNA of CrPV IGR IRES (GenBank accession no. AF218039, nucleotides 6,025–6,216) harboring either ATG (A) or GCT (B) as initiation codon (underlined).

Figure S2. Design of the expression vector EasyXpress pIX3.0 CrPV IGR IRES (A) and pcDNA3.1(+) CrPV IGR IRES (B). The pcDNA3.1 CrPV IGR IRES vector contains a CMV and a T7 promoter whereas the EasyXpress pIX3.0 CrPV IGR IRES construct is limited to the T7 promoter. As a result of the limitation, the use of the pIX3.0-based vector is restricted to protein expression in vitro but enables increased protein expression levels in the CHO-based cell-free protein production system. Different expression levels may be attributed to the different transcriptional products encoding the target protein.

TABLE SI. Oligonucleotide sequences used for DNA template design.

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