Translational_Unit

Part:BBa_K5078006

Designed by: Elise Bennett   Group: iGEM24_UVU-Utah-2   (2024-09-28)
Revision as of 19:14, 28 September 2024 by Registry (Talk | contribs)

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PL1 Transcriptional Unit for NosZ

pL1-NosZ-D.denit (pL1-D.den) is the combination of the Psad Promoter+5' UTR (BBa_K3002001), NosZ D. denitrificans the nitrous oxide reductase gene from Dechloromonas denitrificans codon optimized for expression in Chlamydomonas reinhardtii (BBa_K5078000), the N-terminator sequence of the fluorescent reporter gene mCherry (BBa_K4770013), and the Psad Terminator (BBa_k3002002). pL1-D.den is the nitrogen half of our completed nutrient uptake plasmid (BBa_K5078009). pL1-D.den is similar to our other nitrogen uptake plasmid pL1- NosZ-P.stuzeri (BBa_K5078005) in that they are both responsible for altering the nitrogen pathway of a host bacterium by forcing the host bacterium to undergo the final step of denitrification, the reduction of nitrous oxide (N₂O) into dinitrogen (N₂). By encoding for nitrous oxide reductase, which will act as the catalysis of a copper-dependent two electron reduction of N₂O into water and N₂ [1]. Additionally this has the benefit of preventing the greenhouse gas N₂O from entering the atmosphere and theoretically should cause the host bacterium to uptake more nitrogen from their environment. The difference between them is that pL1-D.den has a higher affinity for N₂O than pL1-NosZ-P-stuzeri [2]. We chose to use both,in different final level 2 plasmid builds, as we were unsure how they would function in Chlamydomonas reinhardtii.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 2941
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2941
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 2941
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 2941
  • 1000
    COMPATIBLE WITH RFC[1000]


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