Composite

Part:BBa_K4630402

Designed by: Lei Bai   Group: iGEM23_WHU-China   (2023-10-08)


N20 screening-LacZ

We employed LacZ as the indicator of successful editing and constructed this composite part: N20s screening-LacZ. The part contains an sgRNA targeting sequence inserted in the LacZ sequence (LazC-N20s: BBa_K4630400), an sgRNA scaffold with a Golden Gate site(14bp) upstream, and a homologous arm to facilitate recombination. The scaffold can load a specific N20 sequence (20nts specificity-determining sequence) and conduct targeting fragment knockout, thereby driving LacZ expression. Users can randomly load a batch of different N20 sequences onto the composite part and identify efficient ones by sequencing blue colonies.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 318
    Illegal EcoRI site found at 2178
    Illegal XbaI site found at 347
    Illegal XbaI site found at 1259
    Illegal XbaI site found at 2207
    Illegal SpeI site found at 835
    Illegal PstI site found at 375
    Illegal PstI site found at 2235
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 318
    Illegal EcoRI site found at 2178
    Illegal NheI site found at 1358
    Illegal NheI site found at 1869
    Illegal NheI site found at 1879
    Illegal NheI site found at 1938
    Illegal SpeI site found at 835
    Illegal PstI site found at 375
    Illegal PstI site found at 2235
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 318
    Illegal EcoRI site found at 2178
    Illegal BglII site found at 456
    Illegal BamHI site found at 357
    Illegal BamHI site found at 2217
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 318
    Illegal EcoRI site found at 2178
    Illegal XbaI site found at 347
    Illegal XbaI site found at 1259
    Illegal XbaI site found at 2207
    Illegal SpeI site found at 835
    Illegal PstI site found at 375
    Illegal PstI site found at 2235
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 318
    Illegal EcoRI site found at 2178
    Illegal XbaI site found at 347
    Illegal XbaI site found at 1259
    Illegal XbaI site found at 2207
    Illegal SpeI site found at 835
    Illegal PstI site found at 375
    Illegal PstI site found at 2235
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 858
    Illegal BsaI.rc site found at 1620


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