Coding

Part:BBa_K4117088:Design

Designed by: Yueyao Qi,Bojun Xiong   Group: iGEM22_BNU-China   (2022-09-30)


CYP2C19


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 4467
    Illegal XbaI site found at 693
    Illegal SpeI site found at 713
    Illegal PstI site found at 1214
    Illegal PstI site found at 3118
    Illegal PstI site found at 4348
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4467
    Illegal SpeI site found at 713
    Illegal PstI site found at 1214
    Illegal PstI site found at 3118
    Illegal PstI site found at 4348
    Illegal NotI site found at 3275
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4467
    Illegal BglII site found at 4386
    Illegal BamHI site found at 2887
    Illegal BamHI site found at 2994
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 4467
    Illegal XbaI site found at 693
    Illegal SpeI site found at 713
    Illegal PstI site found at 1214
    Illegal PstI site found at 3118
    Illegal PstI site found at 4348
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 4467
    Illegal XbaI site found at 693
    Illegal SpeI site found at 713
    Illegal PstI site found at 1214
    Illegal PstI site found at 3118
    Illegal PstI site found at 4348
  • 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 CYP2C19 gene.

Source

We design the CYP2C19 DNA sequence based on the amino acid sequence according to E.coli codon preference.The amino acid sequence is from UniProt:P11712

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]Gasse A, Vennemann M, Köhler H, Schürenkamp J. Toxicogenetic analysis of Δ9-THC-metabolizing enzymes. Int J Legal Med. 2020 Nov;134(6):2095-2103. doi: 10.1007/s00414-020-02380-3. Epub 2020 Jul 25. PMID: 32712703; PMCID: PMC7578149.