Composite

Part:BBa_K2109106

Designed by: Andy Hudson   Group: iGEM16_Lethbridge   (2016-10-14)


AraC-pBAD-RNAPa

This part encodes RNA polymerase under the control of the arabinose-inducible pBAD promoter. A multiple cloning site was introduced 3' of the RNA polymerase coding sequence in order to generate a C-terminal fusion to a target protein.

Overexpression of the RNA polymerase fusion protein upon arabionse induction was attempted. However, the RNA polymerase fusion protein has a size similar (if not indistinguishable from) to the endogenous RNA polymerase within E. coli. As there is no antibody specific for this specific fusion protein, a Western Blot could not be performed to confirm the levels of and therefore the overexpression of the RNA polymerase fusion protein.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2055
    Illegal BamHI site found at 1144
    Illegal XhoI site found at 2091
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1763
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


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Categories
Parameters
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