Part:BBa_K4201020:Design
CrtE_cytoTDS2__T5αOH_RUBY
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 6848
Illegal PstI site found at 2087
Illegal PstI site found at 5638
Illegal PstI site found at 8395
Illegal PstI site found at 13603 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 6848
Illegal PstI site found at 2087
Illegal PstI site found at 5638
Illegal PstI site found at 8395
Illegal PstI site found at 13603 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 6848
Illegal BglII site found at 790
Illegal BglII site found at 1153
Illegal BglII site found at 2021
Illegal BglII site found at 2956
Illegal BglII site found at 4011
Illegal BglII site found at 5324
Illegal BglII site found at 5572
Illegal BamHI site found at 11815
Illegal XhoI site found at 3601
Illegal XhoI site found at 3868 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 6848
Illegal PstI site found at 2087
Illegal PstI site found at 5638
Illegal PstI site found at 8395
Illegal PstI site found at 13603 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 6848
Illegal PstI site found at 2087
Illegal PstI site found at 5638
Illegal PstI site found at 8395
Illegal PstI site found at 13603
Illegal NgoMIV site found at 1697
Illegal NgoMIV site found at 6997
Illegal NgoMIV site found at 9751
Illegal NgoMIV site found at 10330
Illegal NgoMIV site found at 12043 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1
Illegal BsaI site found at 948
Illegal BsaI site found at 1887
Illegal BsaI site found at 2167
Illegal BsaI site found at 3113
Illegal BsaI site found at 5438
Illegal BsaI site found at 5718
Illegal BsaI.rc site found at 934
Illegal BsaI.rc site found at 1873
Illegal BsaI.rc site found at 2153
Illegal BsaI.rc site found at 3100
Illegal BsaI.rc site found at 5424
Illegal BsaI.rc site found at 5704
Illegal BsaI.rc site found at 6651
Design Notes
Part information and design consideration can be found on respective basic parts pages.
In contrast to other composite parts designed by our group, each individual gene in this assembly has its own promoter and terminator, which allows us to study and compare which combinations of adjoining or separated genes results in the best expression of our target genes.
Gmubi is constitutive promoter native to Glyine max5, while AtHSP is the terminator of a heat shock protein that has shown to promote expression in plants6.
Source
CrtE is a GGPP synthase from Pantoea ananatis LMG 201031.
Gmubi promoters originate from Glycine max5.
AtHSP terminators originate from the Arabidopsis thaliana genome6.
cytoTDS2 is a taxadiene synthase native to Taxus chinensis var. mairei optimized for use in Glycine max2.
T5αOH is a hydroxylase from Taxus baccata that converts taxadiene into taxadiene-5α-ol, a step in the paclitaxel pathway3.
RUBY is a reporter gene from the order Caryophyllales4.
de novo Synthesis was completed by iGem sponsors IDT and Twist Biosciences.
References
1. Majer, E., Llorente, B., Rodríguez-Concepción, M. & Daròs, J.-A. Rewiring carotenoid biosynthesis in plants using a viral vector. Sci. Rep. 7, 41645 (2017).
2. Xiong, X. et al. The Taxus genome provides insights into paclitaxel biosynthesis. Nat. Plants 7, 1026–1036 (2021).
3. Edgar, S. et al. Mechanistic Insights into Taxadiene Epoxidation by Taxadiene-5α-Hydroxylase. ACS Chem. Biol. 11, 460–469 (2016).
4. He, Y., Zhang, T., Sun, H., Zhan, H. & Zhao, Y. A reporter for noninvasively monitoring gene expression and plant transformation. Hortic. Res. 7, 1–6 (2020).
5. De La Torre, C. M. & Finer, J. J. The intron and 5’ distal region of the soybean Gmubi promoter contribute to very high levels of gene expression in transiently and stably transformed tissues. Plant Cell Rep. 34, 111–120 (2015).
6. Nagaya, S., Kawamura, K., Shinmyo, A. & Kato, K. The HSP Terminator of Arabidopsis thaliana Increases Gene Expression in Plant Cells. Plant Cell Physiol. 51, 328–32 (2010).