RBS

Part:BBa_K4432015

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


Synthetic RBS designed for Pseudomonas aeruginosa PhzA2-G2 operon

This part is a synthetic RBS specifically designed for the Pseudomonas aeruginosa PhzA2-G2 operon (BBa_K4432041) using the Salis Lab RBS Library Calculator v2.0 [1-3].

Usage and Biology

This RBS was used to drive the expression of the PhzA2-G2 operon (BBa_K4432041) under the control of the T5 promoter (BBa_K4432000) in the composite part BBa_K4432141.

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

Translation Initiation Rate (au)	:	13004.40755	
dG_total (kcal/mol)	:	-5.233951572	
dG_mRNA_rRNA (kcal/mol)	:	-9.50135139
dG_spacing (kcal/mol)	:	0
dG_stacking (kcal/mol)	:	0
dG_standby (kcal/mol)	:	0,00999999
dG_start (kcal/mol)	:	-2.76
dG_mRNA (kcal/mol)	:	-7.179999828
Accuracy (warnings)	:	none

This RBS was selected fortuitously during the cloning process from a library of 96 RBSes (ASATTHATTTTTMWKGAGGWCAGAT) for which the estimated Translation Initiation Rates (TIR) range from 1285.05 to 577765.38.

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