Coding

Part:BBa_K530002

Designed by: Daniel Wolozny   Group: iGEM11_Johns_Hopkins   (2011-08-01)

CRTI

This part has been improved by the DTU-Denmark 2016 team by removing the illegal restriction site BgIII at bp 204, making the part compatible with the RFC21 standard. Please see BBa_K2117013 for details of the new and improved part.



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

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 204
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1108
    Illegal SapI site found at 508

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

CrtI Plasmid Map.png


Sequencing

CrtI Sequencing 2.png


This is the sequencing for colony 7.

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