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I fear for the Raptors season if those three injured Raptors miss significant time, which it appears they are going to. For every developed SSR, we chose four left and right half-site sequences, L1-L4 and R1-R4, that contained two or three mutations at varied half-site positions. Additionally, Brec1 maintained enhanced specificity for proper half-site position 10, which differs between loxP and loxBTR. We performed Rec-seq on Cre kinfolk Dre38 and VCre39 using half-site libraries based on their target substrates rox and loxV, which differ from loxP at 25% and 46% of non-core positions, respectively (Fig. 4a). Dre and VCre most popular the canonical base at practically every place in their goal sites, just like wild-kind Cre (Figs. To identify potential pseudosites, we searched the human genome for sequences that contained the Tre or Brec1 minimal substrate motif, inferred from positions within each half-site with Rec-seq enrichment values higher than 2. Using the RSAT motif scanner45 and search parameters A14C13NT11NNT8A7T6T5NNNNNNNNT5′A6′T7′NNC10′A11′A12′ for Tre and C11NG9NT7T6A5NNNNNNNNT5′A6′A7′NNC10′NT12′G13′ for Brec1, we recognized eight human genomic off-target substrates per SSR, every containing 6-eleven non-core mismatches (Supplementary Tables four and 5). These candidate pseudosites had been cloned into the EGFP reporter, and Tre and Brec1 activity was assessed in HEK293T cells as described above