Composite

Part:BBa_K5165004

Designed by: David Doyle, Ziyang Yin, Ashley Yu, Amy Wang   Group: iGEM24_Concordia-Shanghai   (2024-10-02)


GFP-linked PHB Depolymerase (IPTG inducible promoter)

A well-characterized inducible promoter is used to express GFP-linked PHB depolymerase (BBa_K5165000). This construct serves as a baseline comparison for BBa_K5165002 in terms of PHB depolymerase production. Baseline PHB depolymerase levels can determine the degree of which the larger construct is compromised due to the implementation of the quorum sensing system and other additional elements to achieve the two-step mechanism.

Fig. 1. Average fluorescence detected for WT cells and engineered cells when induced with 0.0mM, 0.4mM, and 1.0mM IPTG.

E. coli BL21 cells were transformed with the plasmid and induced with different concentrations of IPTG. Greatest fluorescence was detected when cells were induced with 0.4mM of IPTG. This experiment verified PHB depolymerase expression with the fluorescent tags.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 697
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 634
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 662


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