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The redoxilator
- Genetic design
- Applications and perspectives
- Safety considerations
The USER assembly standard
- USER fusion of biobricks
USER fusion primer design software
- Abstract
- Instructions
- Output format
Experimental results
- Results and discussion
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The project
The USER fusion primer design software: PHUSER
(Primer Help for USER)
PHUSER output format
Like the input options, the output from PHUSER is kept simple. First an overview of all needed primers is shown. The output is fairly self-explanatory, although it should be noted that primer consist of a fusion tail (ft) and a binding region (br), easily distinguished from one another by the occurence of the essential uracil. For example:
|--ft--| |-------br-------|
GGGTTAAU CAAATTATAGCCATACAG
IMPORTANT: Note that the entire sequence constitutes the primer. The space is simply added for illustrative purposes, and should be omitted when synthesizing the primers
Just below is the grafic overview of biobricks and primers. The first block illustrates the forward primer for biobrick 1, i.e. binding region to biobrick 1, and fusion tail to the selected USER cassette. The following blocks (the number depends on how many biobricks were entered), illustrate the region at which biobrick 1 is fused to biobrick 2 and so forth. The last 40 bases of biobrick 1 and the first 40 bases of biobrick 2 are displayed, along with the positioning of the fusion tails and binding regions for both reverse and forward primers. The last block of the graphic overview illustrated the last biobrick and its binding region to the selected cassette, diplayed in a similar fashion as described for the first block.
Lastly, the primers are displayed again, only this time with more detailed information, such as TM and GC ratio. More parameters are to be added to the output.
Design your primers with PHUSER here
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Achievements
Redox sensing device
Two novel genes have been designed and synthesized each comprised of 5 genetic elements. Together they function as a device termed the Redoxilator that can sense the internal redox state of a yeast cell, and output a reporter signal. Extensive mathematical modelling was performed to simulate how the construct would operate in vivo.
Biobricks
DNA of several new biobricks have been designed and submitted including a yeast optimized GFP reporter protein, a protein degradation sequence and a fast degradable yeast GFP. (Bronze medal)
We have demonstrated that our USER fusion biobrick works as expected and documented it (silver medal)
USER fusion Assembly standard
A new biobrick assembly standard that allows the rapid construction of multi-part devices have been developed and documented. The assembly standard offers many benefits: All restriction sites are allowed, multiple biobricks can be joined in one step, the result is scar-free making it ideal for protein fusions and more. (Gold medal)
USER-fusion primer design software
A novel and very useful software tool have been developed that can automatically design the optimal primers for USER fusion assembly of 2-9 biobricks, taking several parameters into account.
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