Regulatory

Part:BBa_K1484343:Design

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


P_PiI5, marchantia promoter


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 1792
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1069
    Illegal SpeI site found at 1792
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 128
    Illegal BamHI site found at 765
    Illegal XhoI site found at 816
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 1792
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 1792
  • 1000
    COMPATIBLE WITH RFC[1000]


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 proteins for transporters in the PHT1 family in A. Thaliana. 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. Remy E et al. 2012. The Pht1;9 and Pht1;8 transporters mediate inorganic phosphate acquisition by the Arabidopsis thaliana root during phosphorus starvation. New Phytol. 195(2):356-71 TAIR Loci: At5g43350, At5g43370, At5g43360, At2g38940, At2g32830, At5g43340, At3g54700, At1g20860, At1g76430