Regulatory

Part:BBa_K1484342:Design

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


P_PiI3, marchantia promoter


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1753
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1753
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1753
    Illegal BglII site found at 1718
    Illegal BamHI site found at 1745
    Illegal XhoI site found at 1043
    Illegal XhoI site found at 1942
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1753
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1753
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1691


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 Phosphate Transporter and Purple Acid Phosphatase proteins in A. Thaliana, known respectively as AtPT2 and AtPAP10. 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

Nussaume L et al. 2011. Phosphate Import in Plants: Focus on the PHT1 Transporters. Front Plant Sci. 2: 83. Schachtman D et al. 1998. Phosphorus Uptake by Plants: From Soil to Cell. Plant Physiology 116(2): 447-453. TAIR Locus: AT2G38940 ThaleMine ATG id: AT2G16430