The Salis Lab has recently discovered a new mechanism, called Ribosome Drafting, that controls a mRNA's translation rate. Learn more about it in our latest publication: Espah Borujeni and Salis. Translation Initiation is Controlled by RNA Folding Kinetics via a Ribosome Drafting Mechanism, Journal of the American Chemical Society, 2016

Are you engineering riboswitches? See our latest NAR Breakthrough Article Automated physics-based design of riboswitches from diverse RNA aptamers, Nucleic Acid Research, 2016. Are you engineering bacterial operons? In non-model hosts? Then our latest published work should also interest you: Kushwaha and Salis, A Portable Expression Resource for Engineering Cross-species Genetic Circuits and Pathways, Nature Communications, 2015. and Tian and Salis, A Predictive Biophysical Model of Translational Coupling to Coordinate and Control Protein Expression in Bacterial Operons, Nucleic Acid Research, 2015


RBS Library Calculator
Title


Genomic RBS Sequence [?]
Ribosome Binding Site Sequence: enter an initial ribosome binding site sequence, using A/G/C/T/U. A complete RBS sequence is 35 nucleotides long. If none is selected, a random one will be generated and used as the initial condition.
Genomic Protein CDS [?]
Protein Coding Sequence: enter a nucleotide sequence (at least 50 bp) that encodes a protein, using A/G/C/T/U.
Begins with a start codon (ATG/GTG/TTG/CTG). (required)
 
Max Length of Mutation Box [?]
Maximum Number of Consecutive Degenerate Nucleotides: The number of consecutive mutations to the genome partly determine the efficiency of genome editing via oligo-mediated allelic recombinant (OMAR) or MAGE. Select the maximum number of consecutive nucleotides that may be mutated, compared to the initial RBS sequence. Typical values are from 4 to 20 nucleotides.
RBS Constraints [?]
Ribosome Binding Site Sequence Constraints: enter a degenerate nucleotide sequence that determines which mutations to the RBS are allowable. Insert a N where mutations are desired or A/G/C/T/U where mutations are not desired. (required)
(optional)


Minimum Translation Initiation Rate [?]
RBS Library's Minimum Translation Initiation Rate: select the desired minimum rate of translation initiation within the optimized ribosome binding site library, on a proportional scale from 0.1 to 100,000 or more. This will determine the lowest protein expression level within the library. (required)
Maximum Translation Initiation Rate [?]
RBS Library's Maximum Translation Initiation Rate: select the desired maximum rate of translation initiation within the optimized ribosome binding site library, on a proportional scale from 0.1 to 100,000 or more. This will determine the highest protein expression level within the library. (required)


Target Library Size: x     Low Res    
    High Res
Organism or (16S rRNA) [?]
Organism or 16S rRNA sequence: choose a bacterial species by typing in the first 3 letters of its name and selecting it from the list. Alternatively, you may enter the last 9 nucleotides of the 16S rRNA, using A/G/C/T/U.
(required)
(start typing)

Design Jobs: 2 queued, 5 currently running
For Non-Commercial Use Only. Click here for commercial usage.
When using these results, please reference: Iman Farasat, Manish Kushwaha, Jason Collens, Michael Easterbrook, Matthew Guido, and Howard M. Salis, "Efficient search, mapping, and optimization of multi-protein genetic systems in diverse bacteria", Molecular Systems Biology, v10(6), 2014
Method online since January 10th, 2011.
Have a Question? Our Documentation, Publications, and References may have your answer!
We gratefully acknowledge research funding from the Air Force Office of Scientific Research, the National Science Foundation, the Office of Naval Research, and an Amazon AWS Research Grant.
Computational resources are provided by the AWS Elastic Compute Cloud.