Difference between revisions of "Part:BBa K2483005:Design"

 
 
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===Design Notes===
 
===Design Notes===
−
We put random big spacers (118 bp) inbetween the sgRNA target sites to reduce the amount of homologous recombination caused by the repeats.
+
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 118 bp inbetween the target ste couples to reduce homologous recombination. The recoginition sequences were analyzed with an online tool for efficacy.
  
  

Latest revision as of 08:28, 6 September 2017


sgRNA target site couples facing each other with 6 bp spacer


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 562
  • 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 118 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 purely synthetic.

References