Translational_Unit

Part:BBa_K3288065

Designed by: Sunmin Lee   Group: iGEM19_SIS_Korea   (2019-10-21)


light inducible expression of Ideonella sakaiensis PETase(1-290;WT)

BBa_K3288065 is a composite part in which BBa_K1075044, BBa_K3288007 and BBa_K3288009 are combined.

This part is compatible with RFC10.

Usage and Biology

800px-SIS_Korea_figure6.jpeg

SecretionofPETase.jpeg

In order to build a PET degradation device, we had to increase the secretion of PET degradation enzymes. So we searched for a novel signal peptide candidate through literature search and determined the cleavage site using computational analysis.

In order to release PETase (BBa_K2110800) from E. coli, the endogenous signal peptide was removed and a new signal peptide (NSP4) was attached. As a result, it can be seen that NSP4-PETase was secreted more into the media than PETase (BBa_K2110800).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2171
    Illegal XhoI site found at 4325
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 63
    Illegal NgoMIV site found at 195
    Illegal NgoMIV site found at 289
    Illegal NgoMIV site found at 582
    Illegal NgoMIV site found at 1076
    Illegal NgoMIV site found at 1094
    Illegal NgoMIV site found at 1184
    Illegal AgeI site found at 414
    Illegal AgeI site found at 1542
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1643
    Illegal BsaI.rc site found at 525


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Categories
Parameters
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