Part:BBa_K2333428:Design
UNS pTet mScarlet-I pdt A
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1059
Illegal NheI site found at 1082
Illegal NotI site found at 632 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
This part was designed with the aTc inducible promoter pTet for the pdt-A tagged mScarlet-I reporter combined with the pTet repressor tetR under the control of the medium-weak strength constitutive promoter J23105.
Source
Pdt A was originally generated by mutagenesis from the endogenous Lon degraded tags from the bacteria Mycoplasma florum by Collins et al. 2014 "Tunable Protein Degradation in Bacteria". Pdt A corresponds to Collins et al.'s tag pdt#3. To create pdt A, the amino acid sequence was taken from Collins et al. and was codon optimized for E. coli, then synthesized by IDT.
UNS sequences are from Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2013). Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly.
The sequence for mScarlet-I was obtained and modified from the sequence in the paper by Bindels et al. 2016 "MScarlet: a bright monomeric red fluorescent protein for cellular imaging." The sequence was modified to remove the start codon.
References
[1] Bindels, D. S., Haarbosch, L., Weeren, L. V., Postma, M., Wiese, K. E., Mastop, M., . . . Gadella, T. W. (2016). MScarlet: a bright monomeric red fluorescent protein for cellular imaging. Nature Methods, 14(1), 53-56. doi:10.1038/nmeth.4074
[2] Cameron DE, Collins JJ. Tunable protein degradation in bacteria. Nature Biotechnology. 2014;32(12):1276–1281.
[3] Torella JP, Boehm CR, Lienert F, Chen J-H, Way JC, Silver PA. Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly. Nucleic Acids Research. 2013;42(1):681–689.