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Consistent with previous observations [30] these motifs are present on both strands in direct and inverted repeat orientations.
The database facilitates mining of SSR motifs, repeat orientations and sequence similarities.
To investigate how CRISPR-directed SSBs can be applied to delete large sections of genome in E. coli, we designed sgRNAs to target insertion sequences (IS) in direct repeat orientations and tested for genome deletion.
As a result, consensus sequences were found to be composed of bipartite TGGNA motifs separated by 4 or 5 bp AT-rich sequences arranged in direct, reverse direct, inverted, and everted repeat orientations (Fig. 4B).
Specifically, we clearly demonstrate here that the consensus sequence for the high-affinity AbrB binding is comprised of bipartite TGGNA motifs gapped by a 4- or 5-bp AT-rich sequence arranged in direct, reverse direct, inverted, and everted repeat orientations.
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As such, they had to be mirrored; that is, the binding site is repeated immediately adjacent, but in the opposite strand orientation, to maintain the inverted repeat orientation (Fig. 2B).
Analysis of repeat orientation allowed the detection of 52 direct repeats and 22 inverted repeats.
The CTD repeat motif and tandem repeat orientation are observed in most eukaryotic model organisms, and this configuration evolved early in the history of eukaryotes [ 16].
These two genes are located side by side in an inverted repeat orientation on the same chromosome and share a promoter.
Hairpin RNA (hpRNA), a class of short interfering RNA, is composed of two target sequences in an inverted repeat orientation with a spacer between.
Thus, we consider that the repeat orientation tool presented here will be of critical importance for future CRISPR-based experimental studies.
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