Regulatory

Part:BBa_K1484345:Design

Designed by: Angelina Munabi   Group: iGEM14_Cambridge-JIC   (2014-10-10)

P_PiI1, marchantia promoter


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1661
    Illegal PstI site found at 906
    Illegal PstI site found at 1381
    Illegal PstI site found at 1486
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 906
    Illegal PstI site found at 1381
    Illegal PstI site found at 1486
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 151
    Illegal BglII site found at 643
    Illegal XhoI site found at 1192
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1661
    Illegal PstI site found at 906
    Illegal PstI site found at 1381
    Illegal PstI site found at 1486
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1661
    Illegal PstI site found at 906
    Illegal PstI site found at 1381
    Illegal PstI site found at 1486
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1346
    Illegal BsaI.rc site found at 1881


Design Notes

The removal of illegal restriction sites was considered but not completed due to direct interaction of the DNA with regulatory proteins. Also, it was ensured that the region selected for this part was directly upstream of the first ATG of the gene used to identify the promoter sequence. This was so that the 5' UTR, that is vital in plants, was maintained.


Source

This part was found by screening the genome of the Marchantia polymorpha Cam strain maintained by the Haseloff lab for homologues to a Phosphate Transporter protein in the PHT1 family in A. Thaliana: Pht1;6. Transcription is thought to be induced by re-supply of inorganic phosphate after starvation. Matches to the protein CDS sequence were found using Geneious to perform a tblastn search on the genome scaffolds. Predicted genes that contained hits graded above 30% and with at least 40% congruence to mRNA transcript sequences were shortlisted. The best gene candidates (judged according to number and distribution of hits along its length, and supporting mRNA sequence) formed the basis for our predicted promoters. This part was isolated from a 2kb region upstream of the first ATG of such a gene.

References

Wang L et al. 2011. The Arabidopsis purple acid phosphatase AtPAP10 is predominantly associated with the root surface and plays an important role in plant tolerance to phosphate limitation. Plant Physiol. 157(3):1283-99 Hurley B. 2010. The Dual-Targeted Purple Acid Phosphatase Isozyme AtPAP26 Is Essential for Efficient Acclimation of Arabidopsis to Nutritional Phosphate Deprivation. Plant Physiol. 153(3): 1112–1122. ThaleMine ATG id: AT5G34850 ThaleMine ATG id: AT2G16430