Part:BBa_K5252005

XplA

XplA is a cytochrome P450-like gene [1] that along with XplB catalyses the degradation of RDX (Research Department eXplosive or 1,3,5-Trinitroperhydro-1,3,5-triazine) in Rhodococcus rhodochrous. The degradation mechanism is believed to involve denitration, followed by ring cleavage and mineralization, producing byproducts such as nitrite, formaldehyde, and formate. [2]

Adapted from BBa_K3670004 :

- Codon optimized for E. coli while removing SapI, BsaI, BbsI, PaqCI, Esp3I, FaqI restriction sites using Genscript.

- Addition of 3xFLAG-tag and GS-linker sequence

References

[1] Rylott EL, Jackson RG, Sabbadin F, Seth-Smith HM, Edwards J, Chong CS, Strand SE, Grogan G, Bruce NC. The explosive-degrading cytochrome P450 XplA: biochemistry, structural features and prospects for bioremediation. Biochim Biophys Acta. 2011 Jan;1814(1):230-6. doi: 10.1016/j.bbapap.2010.07.004. Epub 2010 Jul 17. PMID: 20624490.

https://pubmed.ncbi.nlm.nih.gov/20624490/#:~:text=XplA%20is%20a%20cytochrome%20P450%20that%20mediates%20the,that%20is%20fused%20to%20its%20flavodoxin%20redox%20partner.

[2] Seth-Smith HM, Rosser SJ, Basran A, Travis ER, Dabbs ER, Nicklin S, Bruce NC. Cloning, sequencing, and characterization of the hexahydro-1,3,5-Trinitro-1,3,5-triazine degradation gene cluster from Rhodococcus rhodochrous. Appl Environ Microbiol. 2002 Oct;68(10):4764-71. doi: 10.1128/AEM.68.10.4764-4771.2002. PMID: 12324318; PMCID: PMC126434.

https://pubmed.ncbi.nlm.nih.gov/12324318/

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 178
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 178
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 178
  Illegal BglII site found at 963
  Illegal BamHI site found at 85
  Illegal BamHI site found at 1398
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 178
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 178
  Illegal NgoMIV site found at 288
  Illegal NgoMIV site found at 937
  Illegal NgoMIV site found at 1557
  Illegal AgeI site found at 343
  Illegal AgeI site found at 460
  
• 1000
  COMPATIBLE WITH RFC[1000]