Part:BBa_K2150101:Design
Tetracycline resistance protein from Bacteroides fragilis
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 562
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
protein sequence--gene sequence(codon preferance) 3 synonynous mutaions
Source
TetX gene is initially found in the transposons Tn4351 and Tn4400 in the anaerobe Bacteroides fragilis We found the sequences of this tetracycline resistance protein in UniProt Protein Database. According to the codon bias in Escherichia coli, we transform the protein sequence into the gene sequence and had it commercially synthesized. In order to avoid illegal restriction sites, we conduct three synonymous mutations in our sequence.
References
[1] Salis, H. M., Mirsky, E. A. & Voigt, C. A. Automated design of synthetic ribosome binding sites to control protein expression. Nat. Biotechnol. 27, 946–50 (2009).
[2] Espah Borujeni, A., Channarasappa, A. S. & Salis, H. M. Translation rate is controlled by coupled trade-offs between site accessibility, selective RNA unfolding and sliding at upstream standby sites. Nucleic Acids Res. 42, 2646–2659 (2014).
[3] Engler, C., Kandzia, R. & Marillonnet, S. A one pot, one step, precision cloning method with high throughput capability. PLoS One 3, (2008).
[4] Gibson, D. G. et al. Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat. Methods 6, 343–5 (2009).