RBS

Part:BBa_K4294114

Designed by: Aristotelis Anastopoulos   Group: iGEM22_Athens   (2022-09-29)


BCD14

A synthetic ribosome binding site for prokaryotic protein expression, built with the guidance of the RBS calculator and characterized by our team.

Measurment

We characterized this RBS in the genetic context of LuxI. We built a fusion of the first 36 nucleotides of the LuxI synthase with sfGFP to characterize the relative strength of our deployed RBS in the context of the LuxI coding sequence. To quantify this output we measured the fluorescence using microplate reader FlexStation3 (Molecular Devices) with excitation wavelength 485nm and emission wavelength 510nm. We conducted measurements in different time points after the induction with aTc, using different concentrations of the inducer.

BCD14luxI sfGFP iGEM Ath 22.png

'Figure 1: Characterization of BCD14 RBS in the genetic context of luxI'

Athens2022-RBS-COMPARISON.png

'Figure 2: Comparison of all the RBS our team characterized in the genetic context of luxI'

References

[1] Mutalik, V. K., Guimaraes, J. C., Cambray, G., Lam, C., Christoffersen, M. J., Mai, Q. A., Tran, A. B., Paull, M., Keasling, J. D., Arkin, A. P., & Endy, D. (2013). Precise and reliable gene expression via standard transcription and translation initiation elements. Nature methods, 10(4), 354–360. https://doi.org/10.1038/nmeth.2404


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None