Part:BBa_K4432010

Synthetic RBS designed for CrtW

This part is a synthetic RBS specifically designed for the Nostoc punctiforme CrtW (BBa_K4432020) using the Salis Lab RBS Library Calculator v2.0 [1-3].

Usage and Biology

This RBS was used to drive the expression of CrtW (BBa_K4432020) under the control of the T5 (BBa_K4432000) or pLac (BBa_R0010) promoters in the composite parts BBa_K4432120, BBa_K4432220 and BBa_K4432320.

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

Translation Initiation Rate (au)	:	183001.2231
dG_total (kcal/mol)	:	-11.10937396
dG_mRNA_rRNA (kcal/mol)	:	-63.4313505
dG_spacing (kcal/mol)	:	0.005325582
dG_stacking (kcal/mol)	:	0
dG_standby (kcal/mol)	:	0.138149285
dG_start (kcal/mol)	:	-2.76
dG_mRNA (kcal/mol)	:	-55.24000168
Accuracy (warnings)	:	none

This RBS was selected as predicted to be the strongest from a library of 108 RBSes (HAGHAAKGVTAAGTAAGGWGGTATAA) for which the estimated Translation Initiation Rates (TIR) range from 292.31 to 183001.22.

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]