Team:SUSTC-Shenzhen/Parts

From 2014.igem.org

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(Parts Table)
(Our RFC)
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We made a new standard and we built three parts based on that this year.  ([http://parts.igem.org/Part:BBa_K1431201 BBa_K1431201],[http://parts.igem.org/Part:BBa_K1431301 BBa_K1431301],[http://parts.igem.org/Part:BBa_K1431302 BBa_K1431302])
We made a new standard and we built three parts based on that this year.  ([http://parts.igem.org/Part:BBa_K1431201 BBa_K1431201],[http://parts.igem.org/Part:BBa_K1431301 BBa_K1431301],[http://parts.igem.org/Part:BBa_K1431302 BBa_K1431302])
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The BbsI restriction enzyme recognize the sequence 5’ GAAGAC3’ and cut downstream two bases to six bases. If we make two reverse directions based on BbsI site, we can cut a part and insert another one with seamless. (Mechanism shown below)
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The BbsI restriction enzyme recognize the sequence <span style="color: red">5'GAAGAC3'<span style="color: black"> and cut downstream two bases to six bases. If we make two reverse directions based on BbsI site, we can cut a part and insert another one with seamless. (Mechanism shown below)
<center>{{SUSTC-Image|wiki/images/a/ab/Our_RFC.png}}</center>
<center>{{SUSTC-Image|wiki/images/a/ab/Our_RFC.png}}</center>

Revision as of 23:01, 17 October 2014

Team SUSTC-Shenzhen

Parts

Our devotion to the community, in reality.

Contents


Our RFC

We made a new standard and we built three parts based on that this year. ([http://parts.igem.org/Part:BBa_K1431201 BBa_K1431201],[http://parts.igem.org/Part:BBa_K1431301 BBa_K1431301],[http://parts.igem.org/Part:BBa_K1431302 BBa_K1431302])

The BbsI restriction enzyme recognize the sequence 5'GAAGAC3' and cut downstream two bases to six bases. If we make two reverse directions based on BbsI site, we can cut a part and insert another one with seamless. (Mechanism shown below)

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Fig.1 The Machanism of Our RFC


Let us make the above sequence as an example. If you want to insert a coding sequence between promoter and polyA, the prefix of forward primer you design must contain GAAGACNNtaaaNNNN and reverse must contain GAAGACNNagttNNNN. The middle two Ns would be any bases for them will be cut off and the four Ns should be begins or ends of the coding sequence. You can get parts by above primers and digest by BbsI restriction enzyme. Then you can insert the parts into promoter and polyA seamless. The PCR product must be such sequence, including the protect bases, shows below.

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Fig.2 PCR Product Sequence

Favorite Parts

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TRE 3G promoter + SV40 PolyA([http://parts.igem.org/Part:BBa_K1431301 BBa_K1431301])would be the best parts what we designed this year and also be our favorite parts.

Parts Table

<groupparts>iGEM2014 SUSTC-Shenzhen</groupparts>

Maintained by the iGEM team SUSTC-Shenzhen.

Licensed under CC BY 4.0.