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Part:BBa_K4244015:Design

Designed by: Arthur Vancolen   Group: iGEM22_Wageningen_UR   (2022-10-08)


dCas9 double repression system


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 33
    Illegal EcoRI site found at 1038
    Illegal EcoRI site found at 2410
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 33
    Illegal EcoRI site found at 1038
    Illegal EcoRI site found at 2410
    Illegal NheI site found at 2169
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 33
    Illegal EcoRI site found at 1038
    Illegal EcoRI site found at 2410
    Illegal BamHI site found at 4448
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 33
    Illegal EcoRI site found at 1038
    Illegal EcoRI site found at 2410
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 33
    Illegal EcoRI site found at 1038
    Illegal EcoRI site found at 2410
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Each protein sequence contains a terminator, so that there is no overlap in expression. For all RBS sequences a medium RBS was chosen, so that the expression of dCas9 would not be too high. Further in the plasmid rhaR should be present when you work with a rhamnose titratable strain. Additionally, the copynumber of this plasmid should be comparable or lower than the three gene plasmid (BBa_K4244024).


Source

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References