Difference between revisions of "Part:BBa K530001"
 
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This part has been improved by the DTU-Denmark 2016 team by removing the illegal restriction site AgeI at bp 202, making the part compatible with the RFC25 standard. Please see <a href="https://parts.igem.org/cgi/partsdb/jumper.cgi?search=BBa_K2117012">BBa_K2117012</a> </html> for details of the new and improved part.
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Enzyme in the pathway required for B-Carotene Synthesis. This enzyme is named Geranylgeranyl Diphosphate Synthase. This sequence was taken from a WT strain of xanthophyllomyces dendrorhous. It catalyzes both the conversion of Geranylgeranyl diphosphate to Phytoene and Lycopene to B-Carotene.  
 
Enzyme in the pathway required for B-Carotene Synthesis. This enzyme is named Geranylgeranyl Diphosphate Synthase. This sequence was taken from a WT strain of xanthophyllomyces dendrorhous. It catalyzes both the conversion of Geranylgeranyl diphosphate to Phytoene and Lycopene to B-Carotene.  

Latest revision as of 21:30, 21 October 2016

CRTE


This part has been improved by the DTU-Denmark 2016 team by removing the illegal restriction site AgeI at bp 202, making the part compatible with the RFC25 standard. Please see BBa_K2117012 for details of the new and improved part.



Enzyme in the pathway required for B-Carotene Synthesis. This enzyme is named Geranylgeranyl Diphosphate Synthase. This sequence was taken from a WT strain of xanthophyllomyces dendrorhous. It catalyzes both the conversion of Geranylgeranyl diphosphate to Phytoene and Lycopene to B-Carotene.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 335
    Illegal XhoI site found at 34
    Illegal XhoI site found at 166
    Illegal XhoI site found at 985
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 202
  • 1000
    COMPATIBLE WITH RFC[1000]


Characterization of performance

Below we can see the beta-carotene production levels on a per cell basis for different combinations of parts involved in beta-carotene synthesis. These are compared with a wild type control. tHMG1 is used to funnel more initial substrate into the pathway and although useful, was not submitted as a part. 2I indicates that two copies of crtI were used.

Vitachar.PNG

Plasmid Map

CrtE Plasmid Map.png

Sequencing

CrtE Sequencing 2.png

This is the sequencing for colony 4.