Composite

Part:BBa_K4630403

Designed by: Zixuan Ding   Group: iGEM23_WHU-China   (2023-10-08)


N20 screening-EGFP

We employed EGFP as the indicator of successful editing and constructed this composite part: N20s screening-LacZ. The part contains an sgRNA targeting sequence inserted in the EGFP sequence (EGFP-N20s: BBa_K4630401), 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 EGFP expression. Users can randomly load a batch of different N20 sequences onto the composite part and identify efficient ones by sequencing green colonies.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1394
    Illegal SpeI site found at 970
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1493
    Illegal NheI site found at 2004
    Illegal NheI site found at 2014
    Illegal NheI site found at 2073
    Illegal SpeI site found at 970
    Illegal NotI site found at 541
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 173
    Illegal XhoI site found at 534
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1394
    Illegal SpeI site found at 970
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1394
    Illegal SpeI site found at 970
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 993
    Illegal BsaI.rc site found at 1755


[edit]
Categories
Parameters
None