Part:BBa_K4073003:Design
BBa_K4073003 regulates salt tolerance in plants.
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 755
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1911
Illegal BsaI.rc site found at 225
Design Notes
Some design considerations that we had to deal with were the regions of the promoter that we wanted to focus on. For example, both RIA1 and RIA3 were regions of interest, and so we had to decide which one of them would yield the most enhanced salinity mechanisms in the form of NaCl response elements. We ended up choosing the restriction enzymes SpeI to KpnI, encompassing both the RIA1 and the RIA3 regions, for the maximum effect of the mechanisms under theoretical NaCl treatments. Additionally, a weak strength RBS was used so that a minimal expression of the HKT1 gene could be achieved (our other composite parts touch upon the other RBS strengths). Another important aspect to consider is that our design could have had the capability to be altered based on how experimental lab procedures followed through. However, due to the lack of access to a lab, we were unable to test the effects of this composite part on the salinity mechanisms of Arabidopsis Thaliana, and therefore most of our design considerations were centered around research gathered from original papers (https://link.springer.com/content/pdf/10.1007/s00253-019-09733-y.pdf).
Source
The source of the weak strength RBS, HKT1 gene, and terminator all came from the iGEM registry. The RBS is the part labeled “BBa_B0031”, the HKT1 gene is the protein coding sequence of the part labeled "BBa_K2665007”, while the terminator is the part "BBa_B0015." The promoter was derived from the NCBI sequence (https://www.ncbi.nlm.nih.gov/nuccore/1601834299) for the CrGPDH3 salt-inducible promoter.