Composite

Part:BBa_K311002

Designed by: Ethan Johnson, Claudia Schmidt-Dannert, Poonam Srivastava, Ian Windsor   Group: iGEM10_Minnesota   (2010-10-15)


Constitutive lac promoter with downstream EGFP

The mutations were generated to get a constitutive lac promoter. The promoter was cloned in pSB1C3 upstream to the EGFP. The promoter activity was checked by transforming the recombinant plasmid in Lac repressor negative (TOP10) and Lac repressor positive (DH5 alpha pro) E. coli cells. The recombinant cells were grown in the presence of varying concentration of inducer (IPTG) and also in the absence of the inducer. The culture was collected at different time points post induction and promoter activity was measured by measuring the fluorescence intensity under FACs. Both the cell types show constitutive behavior of the promoter.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 86
    Illegal XbaI site found at 104
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 86
    Illegal NotI site found at 988
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 86
    Illegal BglII site found at 243
    Illegal BamHI site found at 249
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 86
    Illegal XbaI site found at 104
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 86
    Illegal XbaI site found at 104
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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