Composite

Part:BBa_K5399008:Design

Designed by: josiazhang josiazhang   Group: iGEM24_NAIS   (2024-09-29)


pAtU6-sgRNA-SBEI-3xFLAG-SV40 NLS-cas9-NLS


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1640
    Illegal PstI site found at 3062
    Illegal PstI site found at 3266
    Illegal PstI site found at 3296
    Illegal PstI site found at 4508
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1640
    Illegal PstI site found at 3062
    Illegal PstI site found at 3266
    Illegal PstI site found at 3296
    Illegal PstI site found at 4508
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1101
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1640
    Illegal PstI site found at 3062
    Illegal PstI site found at 3266
    Illegal PstI site found at 3296
    Illegal PstI site found at 4508
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1640
    Illegal PstI site found at 3062
    Illegal PstI site found at 3266
    Illegal PstI site found at 3296
    Illegal PstI site found at 4508
    Illegal NgoMIV site found at 1928
    Illegal NgoMIV site found at 3032
    Illegal NgoMIV site found at 3105
    Illegal NgoMIV site found at 3590
    Illegal NgoMIV site found at 4499
    Illegal NgoMIV site found at 4968
    Illegal NgoMIV site found at 4987
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The pAtU6 is used to initiate the transcription of sgRNA-SBEI; 3xFLAG is used for the detection and purification of the Cas9 protein; SV40 NLS and NLS are used to target the Cas9 protein to the nucleus for the purpose of cleaving the SBEI gene. The design is aimed at producing sgRNAs that target SBEI. The sgRNAs can bind to and guide the Cas9 protein to the target gene locations, leading to gene silencing.


Source

The part is Composite parts.

References