Part:BBa_K5175036:Design

T7 promoter - tphB-tpaK-T7 terminator

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 1906
  Illegal PstI site found at 2195
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 2462
  Illegal PstI site found at 1906
  Illegal PstI site found at 2195
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 242
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 1906
  Illegal PstI site found at 2195
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1906
  Illegal PstI site found at 2195
  Illegal NgoMIV site found at 289
  Illegal NgoMIV site found at 964
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

It is a composite component consisting of the T7 promoter, T7 terminator, target genes tphB, tpaK. It is responsible for oxidising the diol moiety (two hydroxyl groups) of DCD to a keto group to result in the production of PCA and transporting TPA. TphB is a dehydrogenase that oxidises the diol moiety (two hydroxyl groups) of DCD to a keto group, resulting in the production of PCA. TpaK is the TPA transporter protein.

Source

tphB is from Comamonas sp.E6. tpaK is from Rhodococcus erythropolis.

References

[1] SASOH M, MASAI E, ISHIBASHI S, et al. Characterization of the Terephthalate Degradation Genes of Comamonas sp. Strain E6 [J]. Applied and Environmental Microbiology, 2006, 72(3): 1825-32.

[2] KINCANNON W M, ZAHN M, CLARE R, et al. Biochemical and structural characterization of an aromatic ring-hydroxylating dioxygenase for terephthalic acid catabolism [J]. Proc Natl Acad Sci U S A, 2022, 119(13): e2121426119.

[3] HOSAKA M, KAMIMURA N, TORIBAMI S, et al. Novel tripartite aromatic acid transporter essential for terephthalate uptake in Comamonas sp. strain E6 [J]. Appl Environ Microbiol, 2013, 79(19): 6148-55.