Difference between revisions of "Part:BBa K4121067"

 
 
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<partinfo>BBa_K4121067 short</partinfo>
  
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The part is called "pCCW12-PaCrtB-tPRM9". We use pCCW12 as the promoter, tPRM9 as the terminator and the CDS of this part is PaCrtB (<i>Pantoea agglomerans</i> phytoene desaturase). The three basic parts above make up a new transcription unit,It has the function to catalyze the conversion of heterologous geranylgeranyl diphosphate(GGPP) to phytoene.
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===Design Notes===
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We used the "Goldengate" method for the ligation of the basic parts, We introduced recognition sites and cleavage sites for the type IIs restriction endonuclease BsmBI at both ends of the Basic Parts to ensure that the Basic Parts could be ligated successfully, and changed the location of the recognition sites and cleavage sites so that the BsmBI would not cut repeatedly.
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To avoid homologous recombination in Saccharomyces cerevisiae, we substitute different promoters and terminators.
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Please refer to" Design" part of the Wiki for detailed experimental design.
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===Source===
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We get sequences of the promoter from Eukaryotic promoter database(EPD), the sequences of CDS and the terminator from National Center for Biotechnology Information(NCBI).
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===References===
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None
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K4121067 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K4121067 parameters</partinfo>
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Latest revision as of 10:25, 12 October 2022

Expression cassette of PaCrtB with constitutive promoter

The part is called "pCCW12-PaCrtB-tPRM9". We use pCCW12 as the promoter, tPRM9 as the terminator and the CDS of this part is PaCrtB (Pantoea agglomerans phytoene desaturase). The three basic parts above make up a new transcription unit,It has the function to catalyze the conversion of heterologous geranylgeranyl diphosphate(GGPP) to phytoene.

Design Notes

We used the "Goldengate" method for the ligation of the basic parts, We introduced recognition sites and cleavage sites for the type IIs restriction endonuclease BsmBI at both ends of the Basic Parts to ensure that the Basic Parts could be ligated successfully, and changed the location of the recognition sites and cleavage sites so that the BsmBI would not cut repeatedly. To avoid homologous recombination in Saccharomyces cerevisiae, we substitute different promoters and terminators. Please refer to" Design" part of the Wiki for detailed experimental design.

Source

We get sequences of the promoter from Eukaryotic promoter database(EPD), the sequences of CDS and the terminator from National Center for Biotechnology Information(NCBI).

References

None


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 240
    Illegal XbaI site found at 1105
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 240
    Illegal NheI site found at 1455
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 240
    Illegal BglII site found at 1150
    Illegal BglII site found at 1366
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 240
    Illegal XbaI site found at 1105
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 240
    Illegal XbaI site found at 1105
  • 1000
    COMPATIBLE WITH RFC[1000]