Team:ZJU-China/Solution

From 2014.igem.org

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     <p>After reading the Circuit design and Working process of GeneSocket, you may understand how GeneSocket complete its aim: Circuit construction on chromosome. But is its ability only limited in constructing circuit like this? </p>
     <p>After reading the Circuit design and Working process of GeneSocket, you may understand how GeneSocket complete its aim: Circuit construction on chromosome. But is its ability only limited in constructing circuit like this? </p>
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<p><big><big><big>Of course not!</big></big></big></P>
<p><big><big><big>Of course not!</big></big></big></P>
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<img src="https://static.igem.org/mediawiki/2014/5/51/Gfpmode1.jpg"/ align ="right" width="350" ></p>
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<p><b><big>MODE 2 promoter choice</big></b><br />
<p><b><big>MODE 2 promoter choice</big></b><br />
The main propose of promoter BBa_J23110 in our socket is not to promote insertion parts but to guarantee P2 happen. For your own circuit, you may want to use more abundant promoters. So we reserved a homology region in the front of BBa_J23110. If you choose to use your own promoter or have other special design that don’t compatible with socket promoter, you can just choose this region as Left region of insertion fragments.
The main propose of promoter BBa_J23110 in our socket is not to promote insertion parts but to guarantee P2 happen. For your own circuit, you may want to use more abundant promoters. So we reserved a homology region in the front of BBa_J23110. If you choose to use your own promoter or have other special design that don’t compatible with socket promoter, you can just choose this region as Left region of insertion fragments.
<img src="https://static.igem.org/mediawiki/2014/c/cf/ZJU_tool_ch2.jpg"/ align ="right" width="350">
<img src="https://static.igem.org/mediawiki/2014/c/cf/ZJU_tool_ch2.jpg"/ align ="right" width="350">
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<p><b><big>MODE 3 terminator</big></b><br />
<p><b><big>MODE 3 terminator</big></b><br />
If the inserted circuit contains a terminator, it will terminate the downstream expression of socket. So you must link the circuit with a new part, BBa_k1433010. It contains a BBa_J23110 in itself thus guarantee P2 happen.
If the inserted circuit contains a terminator, it will terminate the downstream expression of socket. So you must link the circuit with a new part, BBa_k1433010. It contains a BBa_J23110 in itself thus guarantee P2 happen.

Revision as of 00:01, 18 October 2014

 

After reading the Circuit design and Working process of GeneSocket, you may understand how GeneSocket complete its aim: Circuit construction on chromosome. But is its ability only limited in constructing circuit like this?


Of course not!

We believe GeneSocket has the potential to construct innumerable circuits. But during the construction process, different circuit should have different strategies. It’s not that complicate as it sounds, we even build two standard parts (BBa_K1433009 & BBa_K1433010) to help you face most cases. Our software tools GS-BOX is also compatible with the mode we discuss bellow. Still, we can’t say all situation are included in this discussion, but base on our basic design and principle of GeneSocket, you can come up with best strategies by your own creativity.

Basic principle of GeneSocket P1. Inserted parts must link with standard parts.(why?)
P2. The direct consequence of recombination must be the expression of Int.

Based on this two principles, let’s discuss three insertion modes. This three modes can combine most regular circuit thus answer lots of question of construction!

MODE 1 Simple parts
If we just want to insert a simple part like BBa_E0040. We can link it to BBa_k1433009, who just added a inverted biterminator unit to guarantee the next recombination happen.





MODE 2 promoter choice
The main propose of promoter BBa_J23110 in our socket is not to promote insertion parts but to guarantee P2 happen. For your own circuit, you may want to use more abundant promoters. So we reserved a homology region in the front of BBa_J23110. If you choose to use your own promoter or have other special design that don’t compatible with socket promoter, you can just choose this region as Left region of insertion fragments.






MODE 3 terminator
If the inserted circuit contains a terminator, it will terminate the downstream expression of socket. So you must link the circuit with a new part, BBa_k1433010. It contains a BBa_J23110 in itself thus guarantee P2 happen.

With the methods that we have used above, we can solve most problems when we want to insert a gene-circuit into the chromosome. But sometimes the situation may be more complex. We cannot list all of the situations for you, but we have given you the tool and the idea to solve a new problem. Join the designers’ team and enjoy it!