Difference between revisions of "Part:BBa K4239001:Design"

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===Design Notes===
 
===Design Notes===
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Every Igem restriction sites (EcoR1, Xba1, Spe1 and Pst1) were removed from the part.
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Codon optimization is done for E.coli and D.solani.
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<p>The Igem restriction site PstI was removed into the <i>iluxC</i> part, to create <i>fiatluxC</i>. This mutation does not change the protein sequence.</p>
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<p><i>iluxC</i> had mutations from <i>luxC</i> (N10T, N59D, E74D, S256P, M355T, N360D). They change the protein sequence, in order to create a system that produced more light. The first and the last letters correspond to the Amino Acid before and after mutation, and the number indicates its position in the protein. These mutations were error-prone PCR induced according to Gregor et al.’s study in 2018.</p>
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===Source===
 
===Source===
  
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fiatluxCD is made up of fiatluxC and fiatluxD.
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<p>The source of fiatluxC is synthesis. This part was synthetized from the nucleotidique sequence according to Gregor et al.’s study in 2018.</p>
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<p>This part was synthetized directly with fiatluxD and the promoter BBa_J23117.</p>
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===References===
 
===References===
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<p>Gregor C, Gwosch KC, Sahl SJ, Hell SW. Strongly enhanced bacterial bioluminescence with the ilux operon for single-cell imaging. Proc Natl Acad Sci U S A. 2018 Jan 30;115(5):962-967. doi: 10.1073/pnas.1715946115. Epub 2018 Jan 16. PMID: 29339494; PMCID: PMC5798359.</p>
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Latest revision as of 23:53, 11 October 2022


Enhanced luciferase subunits fiatluxC


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1311


Design Notes

The Igem restriction site PstI was removed into the iluxC part, to create fiatluxC. This mutation does not change the protein sequence.

iluxC had mutations from luxC (N10T, N59D, E74D, S256P, M355T, N360D). They change the protein sequence, in order to create a system that produced more light. The first and the last letters correspond to the Amino Acid before and after mutation, and the number indicates its position in the protein. These mutations were error-prone PCR induced according to Gregor et al.’s study in 2018.

Source

The source of fiatluxC is synthesis. This part was synthetized from the nucleotidique sequence according to Gregor et al.’s study in 2018.

This part was synthetized directly with fiatluxD and the promoter BBa_J23117.

References

Gregor C, Gwosch KC, Sahl SJ, Hell SW. Strongly enhanced bacterial bioluminescence with the ilux operon for single-cell imaging. Proc Natl Acad Sci U S A. 2018 Jan 30;115(5):962-967. doi: 10.1073/pnas.1715946115. Epub 2018 Jan 16. PMID: 29339494; PMCID: PMC5798359.