Composite

Part:BBa_K3726011

Designed by: Santiago Barragán Ariza   Group: iGEM21_MADRID_UCM   (2021-10-18)
Revision as of 17:58, 19 October 2021 by Sbarraga (Talk | contribs)


CDS_Lv0_BOH_3

This composite part corresponds to a phytobrick compatible MoClo Lv.0 CDS element. This part is an artificial operon designed to express the enzymes CDS_NphT7 (BBa_K3726000), CDS_PhaB^173S (BBa_K3726001), CDS_Ccr (BBa_K3726002), CDS_PhaJ (BBa_K3726003), CDS_PduP (BBa_K3726004), CDS_Slr1192 (BBa_K3726005).

Within this artificial operon each CDS element is preceded by an in-silico designed RBS sequence using DeNovo DNA software. Only the first coding sequence of the operon is not preceded by an RBS sequence which must be assembled during the level 1 MoClo reaction for the expression of the operon in a level 1 transcriptional unit.

This part has been used within the iGEM MADRID_UCM 2021 iGEM team for the expression of a butanol biosynthesis pathway.

The relative transcription initiation strengths of each RBS in the operon context has been calculated with DeNovo DNA Software. Results are depicted in the graphic below.

For BOH3

<img class="img-fluid" src="https://static.igem.org/mediawiki/parts/thumb/6/6e/T--MADRID_UCM--RBSBOH3.png/800px-T--MADRID_UCM--RBSBOH3.png&sa=D&source=docs&ust=1634668121947000&usg=AOvVaw22VXNiWjll0c5hDBbQGkVd" alt="DESCRIPCIÓN" data-aos="fade-up">

As can be observed, the PduP and Slr1192 internal RBS sequences have been designed to pose a moderate strength. Since this part corresponds with a fully functional n-butanol biosynthesis, relative expression of each enzyme required for optimal pathway functioning is controlled by their internal RBS.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 461
    Illegal PstI site found at 366
    Illegal PstI site found at 614
    Illegal PstI site found at 1915
    Illegal PstI site found at 4930
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 461
    Illegal NheI site found at 3936
    Illegal NheI site found at 4164
    Illegal PstI site found at 366
    Illegal PstI site found at 614
    Illegal PstI site found at 1915
    Illegal PstI site found at 4930
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 461
    Illegal BglII site found at 5275
    Illegal BglII site found at 5392
    Illegal BamHI site found at 1998
    Illegal BamHI site found at 2337
    Illegal XhoI site found at 3270
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 461
    Illegal PstI site found at 366
    Illegal PstI site found at 614
    Illegal PstI site found at 1915
    Illegal PstI site found at 4930
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 461
    Illegal PstI site found at 366
    Illegal PstI site found at 614
    Illegal PstI site found at 1915
    Illegal PstI site found at 4930
    Illegal NgoMIV site found at 105
    Illegal NgoMIV site found at 205
    Illegal NgoMIV site found at 511
    Illegal NgoMIV site found at 2593
    Illegal AgeI site found at 883
    Illegal AgeI site found at 2121
  • 1000
    COMPATIBLE WITH RFC[1000]


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