Plasmid
CueO-PET28

Part:BBa_K4232008

Designed by: Songyuan Kong   Group: iGEM22_BIT-China   (2022-10-03)


A plasmid used to catalyze oxygen and reducing substances

This is a plasmid made of laccase with pet28a for expression in the introduction of E. coli，it is used to deplete oxygen within cells. Laccase, known as multicopper oxidase CueO,The specific working principle is show in the BBa_K4232000.The specific construction of the laccase plasmid is shown in Figure 1. Since the expression of laccase is affected by copper ions, we added copper ions during the laccase culture process, and the specific difference in fluorescence intensity is shown in Figure 2.

figure 1.Laccase plasmid diagram
figure 2.Difference in fluorescence intensity after addition of copper ions
















Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1751
    Illegal XbaI site found at 1894
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1751
    Illegal NheI site found at 1790
    Illegal NotI site found at 1724
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1751
    Illegal BglII site found at 1960
    Illegal BamHI site found at 1757
    Illegal XhoI site found at 1717
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1751
    Illegal XbaI site found at 1894
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1751
    Illegal XbaI site found at 1894
    Illegal NgoMIV site found at 225
    Illegal NgoMIV site found at 1992
    Illegal NgoMIV site found at 3580
    Illegal NgoMIV site found at 3740
    Illegal NgoMIV site found at 6783
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 4657


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Categories
Parameters
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