Composite

Part:BBa_K4591013

Designed by: XinTong Shen   Group: iGEM23_ZJUT-China   (2023-10-08)


T7-DepoPETase-T500

Usage and Biology

This circuit contains the coding sequences of P43, DepoPETase, and MHETase. The expression of enzyme genes related to this route is controlled by the frequently expressed promoter P43, which degrades PET into TPA and achieves the preliminary degradation of PET.

Basic components of the circuit

P43：P43is a constitutive promoter that constitutively expresses the P43 protein in B.subtilis.

DepoPETase：DepoPETase is the best mutant obtained by researchers using a novel high-throughput screening method based on fluorescence detection for directed evolution of IsPETase.

MHETase：MHETase is discovered from Ideonella sakaiensis 201-F6, it is a enzyme which can hydrolyze MHET, which is the main product of PETase when hydrolyzing PET.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 469
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 239
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 25


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Categories
Parameters
None