Part:BBa_K1484346:Design
P_SuI1, marchantia promoter
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1322
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1322
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1116
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1322
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1322
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 358
Illegal BsaI site found at 1113
Illegal BsaI.rc site found at 113
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 Sulphate Transporter proteins in A. Thaliana known as SULTR1;1 and SULTR1;2. Transcription is thought to be induced when sulphur is limiting. 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
Hawkesford M. 2003. Transporter gene families in plants: the sulphate transporter gene family — redundancy or specialization? Physiologia Plantarum 117 (2): 155–163. Takahashi H. 2000. The roles of three functional sulphate transporters involved in uptake and translocation of sulphate in Arabidopsis thaliana. Plant J. 3(2):171-82. ThaleMine ATG id: AT4G08620 TAIR Locus: AT1G78000