Design and cloning of targets in the pUC19 and pMRS backbone

The fragments cloned into the pMRS vector were selected from previously published work (Supplementary Sequence S3) (42). For targets and larger fragments incorporated into the pUC19 backbone, oligos or g-blocks were custom-ordered and subsequently digested with BamHI and EcoRI enzymes before being cloned. The validity of these clones was confirmed through restriction analysis and Sanger sequencing (Supplementary Sequence S4–S6). These verified clones were employed to evaluate PG-T7EI activity.