Composite

Part:BBa_K4887021:Design

Designed by: Duoqing LIN   Group: iGEM23_Shanghai-BioX   (2023-09-19)


Backbone of IbGBSSI knockout system


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 2106
    Illegal PstI site found at 3528
    Illegal PstI site found at 3732
    Illegal PstI site found at 3762
    Illegal PstI site found at 4974
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2106
    Illegal PstI site found at 3528
    Illegal PstI site found at 3732
    Illegal PstI site found at 3762
    Illegal PstI site found at 4974
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1567
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 2106
    Illegal PstI site found at 3528
    Illegal PstI site found at 3732
    Illegal PstI site found at 3762
    Illegal PstI site found at 4974
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 2106
    Illegal PstI site found at 3528
    Illegal PstI site found at 3732
    Illegal PstI site found at 3762
    Illegal PstI site found at 4974
    Illegal NgoMIV site found at 1157
    Illegal NgoMIV site found at 1176
    Illegal NgoMIV site found at 2394
    Illegal NgoMIV site found at 3498
    Illegal NgoMIV site found at 3571
    Illegal NgoMIV site found at 4056
    Illegal NgoMIV site found at 4965
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part contains: a transcription of a Cas9 protein gene with a nuclear localization signal (NLS, BBa_K4887004) controlled by the promoter pAtUBQ (BBa_K4887008), and a transcription of the sgRNA (BBa_K4887002) controlled by the promoter of the Arabidopsis pAtU6 (BBa_K4887007).


Source

This part is constructed after the sgRNA oligos are ligated into the digested plasmid psgR-Cas9-At using T4 ligase, based on parts: BBa_K4887007, BBa_K4887002,] BBa_K4887008, BBa_K4887003, BBa_K4887004, BBa_K4887005, BBa_K4887004 and BBa_K4887010.


References