Plasmid

Part:BBa_K3978015

Designed by: Haozhen He   Group: iGEM21_HiZJU-China   (2021-10-21)


BL21 vector combined with amoA and hao gene sequence

Nitrosomonas Europaea ATCC 19718 ammonia oxidizing bacteria convert ammonia to hydroxylamine under the action of AMO and subsequently convert hydroxylamine to nitrite under the action of HAO.

According to existing studies, the EE2 biotransformation obviously depends on the oxidation of NH3, that is, the degradation of EE2 needs to be carried out in the environment of NH3. The possible mechanism is that THE HAO oxidizes NH3 to provide electrons for the oxidation of EE2-OH. Ammonia is present in urine and sewage, so degradation can be achieved without the addition of other substances to the reaction system.

We embed the amoA and hao gene sequence into the petDUET vector, which also contains T7 promoter lac operator to conduct the transcription and translation process. We use the B0030 RBS and B0015 terminator. The AmpR pro1moter is concluded for selection. The gene sequence have submitted to website numbered in BBa_K3978011 and BBa_K3978040

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 811
    Illegal EcoRI site found at 2673
    Illegal XbaI site found at 2591
    Illegal PstI site found at 1552
    Illegal PstI site found at 2692
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 811
    Illegal EcoRI site found at 2673
    Illegal PstI site found at 1552
    Illegal PstI site found at 2692
    Illegal NotI site found at 2710
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 811
    Illegal EcoRI site found at 2673
    Illegal BglII site found at 2866
    Illegal BamHI site found at 246
    Illegal BamHI site found at 2667
    Illegal XhoI site found at 2915
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 811
    Illegal EcoRI site found at 2673
    Illegal XbaI site found at 2591
    Illegal PstI site found at 1552
    Illegal PstI site found at 2692
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 811
    Illegal EcoRI site found at 2673
    Illegal XbaI site found at 2591
    Illegal PstI site found at 1552
    Illegal PstI site found at 2692
    Illegal NgoMIV site found at 2885
    Illegal NgoMIV site found at 3232
    Illegal NgoMIV site found at 7909
    Illegal AgeI site found at 270
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 328
    Illegal BsaI.rc site found at 709
    Illegal BsaI.rc site found at 3817
    Illegal SapI.rc site found at 5477


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