Composite
pqq

Part:BBa_K4383048:Design

Designed by: Junxiao Zhang   Group: iGEM22_SCU-China   (2022-10-11)


J23114-B0034-pqqA-pqqB-pqqC-pqqD-pqqE-pqqF


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 872
    Illegal PstI site found at 1537
    Illegal PstI site found at 1745
    Illegal PstI site found at 1805
    Illegal PstI site found at 1915
    Illegal PstI site found at 2221
    Illegal PstI site found at 3213
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal PstI site found at 872
    Illegal PstI site found at 1537
    Illegal PstI site found at 1745
    Illegal PstI site found at 1805
    Illegal PstI site found at 1915
    Illegal PstI site found at 2221
    Illegal PstI site found at 3213
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2370
    Illegal BamHI site found at 1353
    Illegal XhoI site found at 3190
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 872
    Illegal PstI site found at 1537
    Illegal PstI site found at 1745
    Illegal PstI site found at 1805
    Illegal PstI site found at 1915
    Illegal PstI site found at 2221
    Illegal PstI site found at 3213
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 872
    Illegal PstI site found at 1537
    Illegal PstI site found at 1745
    Illegal PstI site found at 1805
    Illegal PstI site found at 1915
    Illegal PstI site found at 2221
    Illegal PstI site found at 3213
    Illegal NgoMIV site found at 215
    Illegal NgoMIV site found at 404
    Illegal NgoMIV site found at 881
    Illegal NgoMIV site found at 967
    Illegal NgoMIV site found at 2015
    Illegal NgoMIV site found at 2068
    Illegal NgoMIV site found at 2539
    Illegal AgeI site found at 1260
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4008
    Illegal BsaI.rc site found at 1981
    Illegal SapI.rc site found at 2866
    Illegal SapI.rc site found at 4624


Design Notes

We cloned the pqq synthesis gene cluster from Klebsiella pneumoniae and combined it with constitutive promoters of different intensities to find the most suitable expression intensity for pqq synthesis.For more information, please view https://2022.igem.wiki/scu-china/design.html.

Source

Klebsiella pneumoniae

References

Zhu W, Klinman JP. Biogenesis of the peptide-derived redox cofactor pyrroloquinoline quinone. Curr Opin Chem Biol. 2020;59:93-103. doi:10.1016/j.cbpa.2020.05.001 Zhu W, Klinman JP. Biogenesis of the peptide-derived redox cofactor pyrroloquinoline quinone. Curr Opin Chem Biol. 2020;59:93-103. doi:10.1016/j.cbpa.2020.05.001 Bonnot F, Iavarone AT, Klinman JP. Multistep, eight-electron oxidation catalyzed by the cofactorless oxidase, PqqC: identification of chemical intermediates and their dependence on molecular oxygen [published correction appears in Biochemistry. 2013 Aug 20;52(33)5704]. Biochemistry. 2013;52(27):4667-4675. doi:10.1021/bi4003315 Evans RL 3rd, Latham JA, Xia Y, Klinman JP, Wilmot CM. Nuclear Magnetic Resonance Structure and Binding Studies of PqqD, a Chaperone Required in the Biosynthesis of the Bacterial Dehydrogenase Cofactor Pyrroloquinoline Quinone. Biochemistry. 2017;56(21):2735-2746. doi:10.1021/acs.biochem.7b00247