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Rec-seq also confirmed previous findings8 that Cre has a pronounced desire in two areas of loxP: half-site positions 5-7 and 10. We noticed 5.0-fold enrichment of the canonical base at place 10, in keeping with studies that Arg259 participates in hydrogen bonding with the C•G base pair at position 10 (refs. Structural and mutagenesis studies23,26,31 advised that mutation of Arg259 would have an effect on specificity at half-site position 10. Indeed, the Arg259→Ala variant confirmed a drop in enrichment at place 10 (from 5.0-fold for wild-kind Cre to 1.1-fold for the mutant), with a modest choice for C or T in the left half-site and G or A in the proper half-site (Fig. 2b, c). For example, in helix B, Rec-seq of the Lys43→Ala mutant resulted in a modest drop in specificity relative to wild-sort Cre, while Met44→Ala resulted in greater preference at positions 5 and 10 (Fig. 2d). In helix D, the Lys86→Ala variant confirmed minimal differences from wild-type Cre (Supplementary Fig. 7a), while the Gln90→Ala variant confirmed total lower enrichment (Fig. 2e). Within the loop between helices J and K, the Arg282→Ala mutant showed higher, quite than decrease, DNA specificity across loxP (Supplementary Fig. 7b). These outcomes show that Cre’s obvious choice at positions 5-7 results from a number of weak or oblique interactions, rather than being strongly decided by residues proximal to those positions

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