RBS

Part:BBa_K4432011

Designed by: Paul Weimer   Group: iGEM22_Evry_Paris-Saclay   (2022-09-26)


Synthetic RBS designed for CrtO

This part is a synthetic RBS specifically designed for the Dietzia psychralcaliphila CrtO (BBa_K4432021) using the Salis Lab RBS Library Calculator v2.0 [1-3].

Usage and Biology

This RBS was used to drive the expression of CrtO (BBa_K4432021) under the control of the T5 (BBa_K4432000) or pLac (BBa_R0010) promoters in the composite parts BBa_K4432121, BBa_K4432221 and BBa_K4432321.

Using the Salis Lab RBS Library Calculator v2.0 [1-3], the predicted features of this RBS are:

Translation Initiation Rate (au)	:	59252.26243
dG_total (kcal/mol)	:	-8.603649283
dG_mRNA_rRNA (kcal/mol)	:	-68.54135111
dG_spacing (kcal/mol)	:	0
dG_stacking (kcal/mol)	:	0
dG_standby (kcal/mol)	:	0,0786
dG_start (kcal/mol)	:	-2,76
dG_mRNA (kcal/mol)	:	-63,08000183
Accuracy (warnings)	:	none

This RBS was selected as predicted to be the strongest from a library of 32 RBSes (GAMCGAYGMWAAGGAGGTATTAS) for which the estimated Translation Initiation Rates (TIR) range from 3653.92 to 59252.26.

References

[1] Reis AC, Salis HM. An automated model test system for systematic development and improvement of gene expression models. ACS synthetic biology (2020) 9: 3145–3156.

[2] Farasat I, Kushwaha M, Collens J, Easterbrook M, Guido M, Salis HM. Efficient search, mapping, and optimization of multi-protein genetic systems in diverse bacteria. Molecular Systems Biology (2014) 10: 731.

[3] Ng CY, Farasat I, Maranas CD, Salis HM. Rational design of a synthetic Entner-Doudoroff pathway for improved and controllable NADPH regeneration. Metabolic Engineering (2015) 29: 86–96.


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]


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Categories
Parameters
None