Difference between revisions of "Part:BBa K4260009"

 
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<partinfo>BBa_K4260009 short</partinfo>
 
<partinfo>BBa_K4260009 short</partinfo>
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[[File:Biobrick9.png|700px]]
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Figure 1. Construct design scheme
  
 
This part contains the necessary elements for the transcription and expression of Isoeugenol Monooxygenase, as well as its transcriptional activator. Isoeugenol Monooxygenase (IsoMo) is an enzyme of Pseudomonas putida IE27, reported by Yamada, Okada, Yoshida & Nagasawa [1] (gene Iso), and has an approximate size of 54 kDa. It carries out the direct conversion of isoeugenol to vanillin by a redox reaction. The transcriptional activator is regulated by the same promoter as IsoMo because of the bidirectional arrangement of the promoter and the location of the coding gene in the antisense chain.  It was decided to direct it to the periplasm in regard to the application it was needed for (please check out BBa_K4260006), using the already existing part BBa_J32015 [2], because of its codon optimization, specific for E. coli.
 
This part contains the necessary elements for the transcription and expression of Isoeugenol Monooxygenase, as well as its transcriptional activator. Isoeugenol Monooxygenase (IsoMo) is an enzyme of Pseudomonas putida IE27, reported by Yamada, Okada, Yoshida & Nagasawa [1] (gene Iso), and has an approximate size of 54 kDa. It carries out the direct conversion of isoeugenol to vanillin by a redox reaction. The transcriptional activator is regulated by the same promoter as IsoMo because of the bidirectional arrangement of the promoter and the location of the coding gene in the antisense chain.  It was decided to direct it to the periplasm in regard to the application it was needed for (please check out BBa_K4260006), using the already existing part BBa_J32015 [2], because of its codon optimization, specific for E. coli.
  
 
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===Usage and Biology===
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===Sources, Usage and Biology===
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==IemR: Transcriptional activator==
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The transcriptional activator is coded by gene IemR, from Pseudomonas nitroreducens Jin1,  (951 bp long) and has a size of about 34.6 kDa. It is specific for the transcription of the Isoeugenol Monooxygenase gene Iem [3]; however, it was decided to employ gene Iso and not IemR because unlike Iem the first one does not lead to the formation of undesired co-products of the reaction like acetaldehyde or vanillic acid [1].
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[[File:IemR.png|700px]]
  
 
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Revision as of 00:22, 10 October 2022


Coding sequence for isoeugenol monooxygenase and transcriptional activator

Biobrick9.png Figure 1. Construct design scheme

This part contains the necessary elements for the transcription and expression of Isoeugenol Monooxygenase, as well as its transcriptional activator. Isoeugenol Monooxygenase (IsoMo) is an enzyme of Pseudomonas putida IE27, reported by Yamada, Okada, Yoshida & Nagasawa [1] (gene Iso), and has an approximate size of 54 kDa. It carries out the direct conversion of isoeugenol to vanillin by a redox reaction. The transcriptional activator is regulated by the same promoter as IsoMo because of the bidirectional arrangement of the promoter and the location of the coding gene in the antisense chain. It was decided to direct it to the periplasm in regard to the application it was needed for (please check out BBa_K4260006), using the already existing part BBa_J32015 [2], because of its codon optimization, specific for E. coli.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 431
    Illegal XhoI site found at 259
    Illegal XhoI site found at 286
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1179
    Illegal NgoMIV site found at 1887
    Illegal NgoMIV site found at 2495
    Illegal AgeI site found at 1711
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1115
    Illegal BsaI.rc site found at 1807