Portion of the Neutral Site 3 of the Synechococcus elongatus UTEX 2973 genome.
- 10Illegal PstI site found at 635
- 12Illegal PstI site found at 635
- 21COMPATIBLE WITH RFC
- 23Illegal PstI site found at 635
- 25Illegal PstI site found at 635
- 1000COMPATIBLE WITH RFC
The length was chosen such that it was at least 750bp and that there were no off-target effects of the gRNA within this sequence. The location of the gRNA target in Neutral Site 3 was also taken into account when choosing this sequence since there could be no overlap between the two. Since we used Kpn1 for cloning the cassette containing the composite part into the CRISPR plasmid (pSL2680 plasmid), this sequence had to be chosen such that were no Kpn1 cut sites within it.
This sequence can be amplified from the S. elongatus UTEX 2973 genome.