https://video.igem.org/videos/watch/59cfb51f-1d34-4f50-93f2-519f9f365d82 LGG already carries a complete native type II-A CRISPR locus. Our construct does not clone a nuclease. It supplies only the guide. The sgRNA cassette is transcribed, the guide folds and loads into the cell's own Cas9, and the complex scans for its PAM. The target is spaC, the adhesin at the pilus tip and the first gene of the operon, cut at base pair 278. This is the cut Pilinator-7 carries, rank 1 of three target sites. 00:00 The promoter fires and the cassette is transcribed 00:07 A single guide RNA, folded and ready 00:15 The target locus, spaCBA srtC1 00:23 PAM interrogation at 5'-NGAAA-3' 00:31 The blunt double strand break Chapter 5 of 7 from the FSU iGEM 2026 project Lilieum. Base pair positions are given in spaC coding sequence coordinates. https://2026.igem.wiki/fsu Tue, 20 Oct 2026 02:55:05 GMT https://validator.w3.org/feed/docs/rss2.html PeerTube - https://video.igem.org https://video.igem.org/client/assets/images/icons/icon-1500x1500.png https://video.igem.org/videos/watch/59cfb51f-1d34-4f50-93f2-519f9f365d82 All rights reserved, unless otherwise specified in the terms specified at https://video.igem.org/about and potential licenses granted by each content's rightholder.