DNA

Part:BBa_K2483006:Design

Designed by: Bryan Nowack   Group: iGEM17_Potsdam   (2017-09-06)


sgRNA target site couples facing each other with 18 bp spacer


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 226
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 606
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

To guarantee that both sgRNAs face each other, we had to make one sgRNA bind to the opposite strand. We also have big spacer of 127 bp inbetween the target ste couples to reduce homologous recombination. The recoginition sequences were analyzed with an online tool for efficacy.


Source

The sequence is completely synthetic.

References