Difference between revisions of "Part:BBa K2739001:Design"
 
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The promoter used in this part is native to phaR and R.eutropha. Sequence of this native promoter could be found in iGEM registry, as a basic parts (BBa_K108014) and being not available. The source of these sequences were recorded as 1. Regulation of phasin expression and polyhydroxyalkanoate (PHA) granule formation in Ralstonia eutropha H16. Microbiology. 2002, 148:2413–2426. and 2. NCBI (with no further information), having no mentioned if the sequence was codon optimised.
 
The promoter used in this part is native to phaR and R.eutropha. Sequence of this native promoter could be found in iGEM registry, as a basic parts (BBa_K108014) and being not available. The source of these sequences were recorded as 1. Regulation of phasin expression and polyhydroxyalkanoate (PHA) granule formation in Ralstonia eutropha H16. Microbiology. 2002, 148:2413–2426. and 2. NCBI (with no further information), having no mentioned if the sequence was codon optimised.
  
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We access the online NCBI data and found out the current available the functional promoter is being 134bp shorter. Therefore we have used the new phaR and promoter sequence as our new template. Due to the excessive GC in the sequence, we have it codon optimised to E.coli and generated the part through the IDT company service.
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We access the online NCBI data and found out the current available the functional promoter is being 134bp shorter . Therefore we have used the new promoter sequence as our new template and generated the part through the IDT company service.
  
  
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ProR Sequence selection based on Reference 1 information.  
 
ProR Sequence selection based on Reference 1 information.  
  
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PhaR sequence codon optimizatied through IDT
 
  
 
===References===
 
===References===
  
 
1. Yamada M, Takahashi S, Okahata Y, Doi Y, Numata K. (2013 ) Monitoring and kinetic analysis of the molecular interactions by which a repressor protein, PhaR, binds to target DNAs and poly[(R)-3-hydroxybutyrate]. AMB Express. 3(1):6.
 
1. Yamada M, Takahashi S, Okahata Y, Doi Y, Numata K. (2013 ) Monitoring and kinetic analysis of the molecular interactions by which a repressor protein, PhaR, binds to target DNAs and poly[(R)-3-hydroxybutyrate]. AMB Express. 3(1):6.

Latest revision as of 01:22, 18 October 2018


The promoter (ProR) + RBS for the phasin autoregulation system (PhaR)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part is consisted PhaR's native promoter. This part was designed to allow the investigation of phaR relation in PHA production through PHA operon +/- phasin.

The promoter used in this part is native to phaR and R.eutropha. Sequence of this native promoter could be found in iGEM registry, as a basic parts (BBa_K108014) and being not available. The source of these sequences were recorded as 1. Regulation of phasin expression and polyhydroxyalkanoate (PHA) granule formation in Ralstonia eutropha H16. Microbiology. 2002, 148:2413–2426. and 2. NCBI (with no further information), having no mentioned if the sequence was codon optimised.

We access the online NCBI data and found out the current available the functional promoter is being 134bp shorter . Therefore we have used the new promoter sequence as our new template and generated the part through the IDT company service.


Source

The ProR promoter is native to R.eutropha (NCBI: AM260479.1).

ProR Sequence selection based on Reference 1 information.


References

1. Yamada M, Takahashi S, Okahata Y, Doi Y, Numata K. (2013 ) Monitoring and kinetic analysis of the molecular interactions by which a repressor protein, PhaR, binds to target DNAs and poly[(R)-3-hydroxybutyrate]. AMB Express. 3(1):6.