Coding

Part:BBa_K4117888:Design

Designed by: He Du,Yue Zhang   Group: iGEM22_BNU-China   (2022-09-30)


UGT1A3


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 4599
    Illegal XbaI site found at 693
    Illegal SpeI site found at 713
    Illegal PstI site found at 1214
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4599
    Illegal SpeI site found at 713
    Illegal PstI site found at 1214
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4599
    Illegal BglII site found at 4257
    Illegal BamHI site found at 2887
    Illegal BamHI site found at 3444
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 4599
    Illegal XbaI site found at 693
    Illegal SpeI site found at 713
    Illegal PstI site found at 1214
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 4599
    Illegal XbaI site found at 693
    Illegal SpeI site found at 713
    Illegal PstI site found at 1214
    Illegal AgeI site found at 3031
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 732


Design Notes

To degrade Δ9-THC, we added a pathway to sense Δ9-THC in front of the UGT1A3 gene.

Source

We design the DNA sequence based on the amino acid sequence according to E·coli codon preference.The amino acid sequence is from UniProt:P35503

anti-THC pmrB gene come from Part:BBa_K4117779.

The pmrC promoter comes from Part:BBa_K1459006.

The pmrA gene comes from Part:BBa_K3611003.

The ompT signal peptide gene comes from Part:BBa_K3805532.

The ompA signal peptide gene comes from Part:BBa_K103006.

T7 terminator gene comes from Part:BBa_B0012.

The rrnB T1 Terminator gene comes from BBa_K3257020.

References

[1] Sinsinbar G, Gudlur S, Metcalf KJ, Mrksich M, Nallani M, Liedberg B. Role of Lipopolysaccharide in Protecting OmpT from Autoproteolysis during In Vitro Refolding. Biomolecules. 2020;10(6):922. Published 2020 Jun 18.Ct

[2] Sugimura K, Nishihara T. Purification, characterization, and primary structure of Escherichia coli protease VII with specificity for paired basic residues: identity of protease VII and OmpT. J Bacteriol. 1988 Dec;170(12):5625-32. doi: 10.1128/jb.170.12.5625-5632.1988. PMID: 3056908; PMCID: PMC211661.

[3] Mazur A, Lichti CF, Prather PL, Zielinska AK, Bratton SM, Gallus-Zawada A, Finel M, Miller GP, Radomińska-Pandya A, Moran JH. Characterization of human hepatic and extrahepatic UDP-glucuronosyltransferase enzymes involved in the metabolism of classic cannabinoids. Drug Metab Dispos. 2009 Jul;37(7):1496-504. doi: 10.1124/dmd.109.026898. Epub 2009 Apr 1. PMID: 19339377; PMCID: PMC2698943.