Part:BBa_K4232008

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.The protein structure of the laccase is shown in the figure 1.

figure 1.Protein structure of laccase

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 2.

figure 2.Laccase plasmid diagram

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 3.

figure 3.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