Composite

Part:BBa_K4201018

Designed by: David Birkhaeuser   Group: iGEM22_CU-Boulder   (2022-10-12)


CrtE_cytoTDS2-MBP_RUBY

This construct is split up into three main gene segments. At the center of each segment lies a gene of interest, in this case CrtE, cytoTDS2-MBP, and RUBY, respectively. Each segment is flanked upstream by the promoter Gmubi and downstream by the terminator AtHSP.

CrtE encodes for a GGPP synthase that converts DMAPP into GGPP1, a precursor in the paclitaxel pathway. Taxadiene synthase II, an enzyme encoded by cytoTDS2, then converts the GGPP into taxadiene2, our target molecule. The code for maltose Binding Protein (MBP) is a solubility tag that stabilizes protein during transport3, that is bordering cytoTDS2.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 4087
    Illegal PstI site found at 2087
    Illegal PstI site found at 3774
    Illegal PstI site found at 5335
    Illegal PstI site found at 6893
    Illegal PstI site found at 12101
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2087
    Illegal PstI site found at 3774
    Illegal PstI site found at 5335
    Illegal PstI site found at 6893
    Illegal PstI site found at 12101
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 790
    Illegal BglII site found at 1153
    Illegal BglII site found at 2021
    Illegal BglII site found at 2956
    Illegal BglII site found at 6827
    Illegal BglII site found at 7762
    Illegal BglII site found at 11664
    Illegal BamHI site found at 10313
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 4087
    Illegal PstI site found at 2087
    Illegal PstI site found at 3774
    Illegal PstI site found at 5335
    Illegal PstI site found at 6893
    Illegal PstI site found at 12101
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 4087
    Illegal PstI site found at 2087
    Illegal PstI site found at 3774
    Illegal PstI site found at 5335
    Illegal PstI site found at 6893
    Illegal PstI site found at 12101
    Illegal NgoMIV site found at 1697
    Illegal NgoMIV site found at 8249
    Illegal NgoMIV site found at 8828
    Illegal NgoMIV site found at 10541
    Illegal AgeI site found at 3923
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1
    Illegal BsaI site found at 948
    Illegal BsaI site found at 1887
    Illegal BsaI site found at 2167
    Illegal BsaI site found at 3114
    Illegal BsaI site found at 6693
    Illegal BsaI site found at 6973
    Illegal BsaI.rc site found at 934
    Illegal BsaI.rc site found at 1873
    Illegal BsaI.rc site found at 2153
    Illegal BsaI.rc site found at 3100
    Illegal BsaI.rc site found at 6679
    Illegal BsaI.rc site found at 6959
    Illegal BsaI.rc site found at 7906


[edit]
Categories
Parameters
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