Part:BBa_K4117008:Design

CYP2C9

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 BamHI site found at 2887
  Illegal BamHI site found at 4377
  
• 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 CYP2C9 gene.

Source

We design the CYP2C9 DNA sequence based on the amino acid sequence according to E. coli codon preference.The amino acid sequence is from NCBI:P33261.4.

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]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.

[2]Watanabe K, Yamaori S, Funahashi T, Kimura T, Yamamoto I. Cytochrome P450 enzymes involved in the metabolism of tetrahydrocannabinols and cannabinol by human hepatic microsomes. Life Sci. 2007 Mar 20;80(15):1415-9. doi: 10.1016/j.lfs.2006.12.032. Epub 2007 Jan 17. PMID: 17303175.

[3]Doohan, P.T., Oldfield, L.D., Arnold, J.C. et al. Cannabinoid Interactions with Cytochrome P450 Drug Metabolism: a Full-Spectrum Characterization. AAPS J 23, 91 (2021). https://doi.org/10.1208/s12248-021-00616-7

[4]Patilea-Vrana GI, Anoshchenko O, Unadkat JD. Hepatic Enzymes Relevant to the Disposition of (-)-∆9-Tetrahydrocannabinol (THC) and Its Psychoactive Metabolite, 11-OH-THC. Drug Metab Dispos. 2019 Mar;47(3):249-256. doi: 10.1124/dmd.118.085548. Epub 2018 Dec 19. PMID: 30567877; PMCID: PMC6374540.

[5]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.

[6]Chen X, Zaro JL, Shen WC. Fusion protein linkers: property, design and functionality. Adv Drug Deliv Rev. 2013 Oct;65(10):1357-69