DNA

Part:BBa_K3971007:Design

Designed by: Misaal Bedi   Group: iGEM21_IISER-Pune-India   (2021-08-10)


Portion of the Neutral Site 1 of the Synechococcus elongatus UTEX 2973 genome.


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 353
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 353
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 353
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 353
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 143


Design Notes

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 1 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, this sequence had to be chosen such that were no Kpn1 cut sites within it.


Source

This sequence can be amplified from the S. elongatus UTEX 2973 genome.

References