Difference between revisions of "Part:BBa K2332021:Design"

 
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===Design Notes===
 
===Design Notes===
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Vibrio fischeri LuxR and Lux operon GenBank: AF170104.1 from which LuxC, D, A, B, E genes were identified and codon optimized for E. coli. The 5kb construct was split into 8 gblock fragments (700-900bp each) and ordered from IDT
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Vibrio fischeri regulatory protein LuxR (luxR) gene, complete cds; LuxICDABEG operon sequences were obtained from Genbank: [https://www.ncbi.nlm.nih.gov/nuccore/5726577 AF170104.1], from which LuxC,D,A,B,E genes were identified and codon optimized for E. coli. Pblrep was designed by Jayaraman P. et al. (2016).
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===Source===
 
===Source===
  
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The 5kb construct is too large to obtain as a single construct so it was split into gBlock fragments and each were designed to have 40bp overlaps to the adjacent fragment for gibson assembly.
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Vibrio fischeri  Genbank: [https://www.ncbi.nlm.nih.gov/nuccore/5726577 AF170104.1]codon optimized for E. coli. The DNA sequence was synthesised by Integrated DNA Technologies (IDT)
  
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===References===
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1. Chong G, Kimyon Ö, Manefield M. Quorum Sensing Signal Synthesis May Represent a Selective Advantage Independent of Its Role in Regulation of Bioluminescence in Vibrio fischeri. PLoS ONE. 2013;8(6):e67443.
  
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===References===
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2. Jayaraman P, Devarajan K, Chua T, Zhang H, Gunawan E, Poh C. Blue light-mediated transcriptional activation and repression of gene expression in bacteria. Nucleic Acids Research. 2016;44(14):6994-7005.

Latest revision as of 00:18, 23 October 2017


Blue light repressible bioluminescence (Pblrep LuxCDABE )


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 2621
    Illegal AgeI site found at 2888
    Illegal AgeI site found at 3416
    Illegal AgeI site found at 3997
    Illegal AgeI site found at 4444
    Illegal AgeI site found at 5322
    Illegal AgeI site found at 5537
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Vibrio fischeri regulatory protein LuxR (luxR) gene, complete cds; LuxICDABEG operon sequences were obtained from Genbank: AF170104.1, from which LuxC,D,A,B,E genes were identified and codon optimized for E. coli. Pblrep was designed by Jayaraman P. et al. (2016).

Source

Vibrio fischeri Genbank: AF170104.1codon optimized for E. coli. The DNA sequence was synthesised by Integrated DNA Technologies (IDT)

References

1. Chong G, Kimyon Ö, Manefield M. Quorum Sensing Signal Synthesis May Represent a Selective Advantage Independent of Its Role in Regulation of Bioluminescence in Vibrio fischeri. PLoS ONE. 2013;8(6):e67443.

2. Jayaraman P, Devarajan K, Chua T, Zhang H, Gunawan E, Poh C. Blue light-mediated transcriptional activation and repression of gene expression in bacteria. Nucleic Acids Research. 2016;44(14):6994-7005.