DNA

Part:BBa_K3570006

Designed by: Anton Mykhailiuk   Group: iGEM20_Toulouse_INSA-UPS   (2020-09-30)
Revision as of 12:21, 22 October 2020 by Cecilia34 (Talk | contribs)


DPP1 upstream homologous sequence

Usage

DPP1 upstream homology arm part shall be used together with DPP1 downstream homology arm part (BBa_K3570007) to target a functional yeast integration locus. When DPP1 up put to 5' of the biobrick together with DPP1 downstream to the 3', the biobrick can be integrated into the S. cerevisiae's genome. It will do homologous recombination within the Diacylglycerol pyrophosphate phosphatase 1 (DPP1) gene.

This sequence was identified from a personal communication with Dr. Gilles Truan.

Experiments

We used BBa_K3570006 and BBa_K3570007 for the integration in the yeast genome of the part BBa_K3570000 to improve the mevalonate pathway. We digested the genome of the transformed yeast with SbfI and EcoRI. As you can see on the figure below, the colonie 2 has the desired digestion profil, which means that BBa_K3570000 part is well integrated in the yeast genome. The integration in the yeast genome is a success. The parts BBa_K3570006 and BBa_K3570007 (for DPP1 homologous sequence) works.


Integration of GGPP insert using DPP1 homologous sequence.png

Figure: Verification of the integration of BBa_K3570000 part into the yeast genome using SbfI and EcoRI.The integration locus is DPP1.

References

  • S. cerevisiae genome, chromosome IV, DPP1 gene. GenBank: CP046084.1

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 50
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 50
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 50
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 50
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None