Part:BBa_K4601106

Synthetic RBS designed for NpHTRII-EnvZ

This part is a synthetic RBS specifically designed for the NpHtrII halobacterial transducer protein fused wih the EnvZ histidine kinase domain (BBa_K4601006) using the Salis Lab RBS Library Calculator v2.0 [1-3].

Usage and Biology

This RBS was used to drive the expression of NpHtrII-EnvZ (BBa_K4601006) under the control of the T7 promoter (BBa_K2150031) in the composite part BBa_K4601206.

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

Translation Initiation Rate (au)	:	9765.62
dG_total (kcal/mol)	:	-4.60
dG_mRNA_rRNA (kcal/mol)	:	-14.71
dG_spacing (kcal/mol)	:	0.0053
dG_stacking (kcal/mol)	:	-0.30
dG_standby (kcal/mol)	:	0.63
dG_start (kcal/mol)	:	-2.76
dG_mRNA (kcal/mol)	:	-12.54
Warnings	:	none

This RBS was selected as predicted to have a Translation Initiation Rate (TIR) of about 10000 au from a library of 96 RBSes (GGGBSTTACTTTCCATCTAAGRAGGTWNGT) for which the estimated TIR range from 102.72 to 36428.08.

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]