Composite

Part:BBa_M50056

Designed by: Lucy Maynard, Doris Mai and Amy Weissenbach   Group: Stanford BIOE44 - S11   (2016-12-11)
Revision as of 05:23, 12 December 2016 by DorisMai (Talk | contribs)


PETase double mutant I208V and R90A full plasmid information

We use Escherichia coli as the chassis organism to express the PETase gene and optimize our codons accordingly. This composite part is made of a rhamnose-inducible promoter, an ampicillin resistance marker, an origin of replication, a secretion tag, a strong RBS, and a PETase gene that contains two mutations. All basic parts except mutated PETase come from DNA 2.0. Wild type PETase shows the ability to hydrolyze Polyethylene terephthalate (PET), commonly used for plastics. The two mutations of PETase are identified by the 2016 iGEM Tianjin team to improve the catalytic activity of PET independently. The PETase gene here also incorporates 6 his tag at the end.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 633
    Illegal PstI site found at 864
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 633
    Illegal PstI site found at 864
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 633
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 633
    Illegal PstI site found at 864
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 633
    Illegal PstI site found at 864
    Illegal NgoMIV site found at 220
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 388


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Categories
Parameters
None