Composite

Part:BBa_K1650007:Design

Designed by: Daniel Stukenberg   Group: iGEM15_Marburg   (2015-09-18)


Composite carrying the Lac-inducible CDI-operon, Lac-inducible RFP and LacI


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 9139
    Illegal PstI site found at 3585
    Illegal PstI site found at 3770
    Illegal PstI site found at 4265
    Illegal PstI site found at 4340
    Illegal PstI site found at 5372
    Illegal PstI site found at 5519
    Illegal PstI site found at 5658
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 9139
    Illegal NheI site found at 947
    Illegal NheI site found at 970
    Illegal NheI site found at 2218
    Illegal NheI site found at 2241
    Illegal PstI site found at 3585
    Illegal PstI site found at 3770
    Illegal PstI site found at 4265
    Illegal PstI site found at 4340
    Illegal PstI site found at 5372
    Illegal PstI site found at 5519
    Illegal PstI site found at 5658
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 9139
    Illegal BglII site found at 2148
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 9139
    Illegal PstI site found at 3585
    Illegal PstI site found at 3770
    Illegal PstI site found at 4265
    Illegal PstI site found at 4340
    Illegal PstI site found at 5372
    Illegal PstI site found at 5519
    Illegal PstI site found at 5658
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 9139
    Illegal PstI site found at 3585
    Illegal PstI site found at 3770
    Illegal PstI site found at 4265
    Illegal PstI site found at 4340
    Illegal PstI site found at 5372
    Illegal PstI site found at 5519
    Illegal PstI site found at 5658
    Illegal NgoMIV site found at 5393
    Illegal NgoMIV site found at 5888
    Illegal NgoMIV site found at 7463
    Illegal NgoMIV site found at 9017
    Illegal AgeI site found at 781
    Illegal AgeI site found at 893
    Illegal AgeI site found at 2827
    Illegal AgeI site found at 2939
    Illegal AgeI site found at 4028
    Illegal AgeI site found at 5202
    Illegal AgeI site found at 5255
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4752
    Illegal BsaI site found at 9583
    Illegal BsaI site found at 10004
    Illegal BsaI.rc site found at 4260
    Illegal BsaI.rc site found at 4527
    Illegal BsaI.rc site found at 13265


Design Notes

BBa_K1650007 was toxic for cells in pSB1C3 backbone, presumably because of the high metabolic burden. In accordance with iGEM Headquarter, we submitted BBa_K1650007 in the low copy pSB4C5 backbone


Source

The CDI-operon was PCR amplified from genomic DNA of E.coli strain EC93. Other parts were taken from the registry.

References