Composite

Part:BBa_K4354008:Design

Designed by: Java Yao   Group: iGEM22_GXU-China   (2022-07-28)


BioBrick used to express enzymes about total nitrogen removal and report gene GFP


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 501
    Illegal EcoRI site found at 986
    Illegal EcoRI site found at 4660
    Illegal PstI site found at 1851
    Illegal PstI site found at 3007
    Illegal PstI site found at 3013
    Illegal PstI site found at 3771
    Illegal PstI site found at 5066
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 501
    Illegal EcoRI site found at 986
    Illegal EcoRI site found at 4660
    Illegal NheI site found at 2987
    Illegal PstI site found at 1851
    Illegal PstI site found at 3007
    Illegal PstI site found at 3013
    Illegal PstI site found at 3771
    Illegal PstI site found at 5066
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 501
    Illegal EcoRI site found at 986
    Illegal EcoRI site found at 4660
    Illegal BglII site found at 2572
    Illegal BamHI site found at 1687
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 501
    Illegal EcoRI site found at 986
    Illegal EcoRI site found at 4660
    Illegal PstI site found at 1851
    Illegal PstI site found at 3007
    Illegal PstI site found at 3013
    Illegal PstI site found at 3771
    Illegal PstI site found at 5066
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 501
    Illegal EcoRI site found at 986
    Illegal EcoRI site found at 4660
    Illegal PstI site found at 1851
    Illegal PstI site found at 3007
    Illegal PstI site found at 3013
    Illegal PstI site found at 3771
    Illegal PstI site found at 5066
    Illegal NgoMIV site found at 4235
    Illegal NgoMIV site found at 4427
    Illegal AgeI site found at 4814
    Illegal AgeI site found at 5431
    Illegal AgeI site found at 5873
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 884
    Illegal BsaI.rc site found at 1696
    Illegal BsaI.rc site found at 5183
    Illegal BsaI.rc site found at 5462
    Illegal BsaI.rc site found at 6567
    Illegal SapI.rc site found at 1839


Design Notes

We will cancel the ampicillin resistance of this vector because it is incompatible with another plasmid we used. We will clone four genes named AMO,HAO,NAR,NIR and a promoter named PBlind on it. As we wished, it can have the function of nitrogen removal in waste water treatent. By testing the expression of GFP, we could know if our bio-brick was engineered successfully.

Source

We synthesize it from HonorGene company according to its sequence and we test the sequence to confirm the accuracy of this part.

References

1.Song Qin, Xu Lei.Advancement of Enzymology of Heterotrophic Nitrification.BIOTECHNOLOGY BULLETIN.DOI:10.13560/j.cnki.biotech.bull.1985.2008.05.009 2.Xu Tao.METABOLIC CHARACTERISTICS OF HETEROTROPHIC NITRIFYING-AEROBIC DENITRIFYING STRAIN DIAPHOROBACTER SP.PD-7.Taiyuan University of Technology in China.2015 May10. 3.Chen Jia-wei，Zhao Pei-jing，Deng Jin-bo，et al.Construction and expression of aiiA with P43 promoter in Bacillus subtilis. Guangdong Agricultural Sciences. 2018, 45(11): 21-27.