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Team:UGent/CloneManager
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<p> To perform in silico analysis, we used <b>Clone Manager</b>. All plasmids were first implemented into the software, and then used to help with cloning simulation, restriction, PCR etc. </p> | <p> To perform in silico analysis, we used <b>Clone Manager</b>. All plasmids were first implemented into the software, and then used to help with cloning simulation, restriction, PCR etc. </p> | ||
<h1> Plasmid containing construct for chromosomal evolution</h1> | <h1> Plasmid containing construct for chromosomal evolution</h1> | ||
<h2>pTGD ccdA Pmb1 BCD7 GFP CmFRT</h2> | <h2>pTGD ccdA Pmb1 BCD7 GFP CmFRT</h2> | ||
| - | <center><img src="https://static.igem.org/mediawiki/2013/e/e9/UGent_2013_PTGD-ccdA-Pmb1-BCD7-GFP-CmFRT.png" width="700" /></center> | + | <p> This plasmid contains the <i>ccdA</i> antitoxin and the reporter gene <i>gfp</i>. The homologous regions are necessary to perform <a href="https://2013.igem.org/Team:UGent/LiteratureStudy" target="_blank"> Chemically Inducible Chromosomal evolution (CIChE)</a>. If wanted, the removal of the chloramphenicol resistance cassette between the FTR sites can be done using the method of <a href="https://static.igem.org/mediawiki/2013/1/18/UGent_2013_Datsenko-Wanner.jpg" target="_blank"> Datsenko & Wanner [PNAS 2000]</a>.</p> |
| - | <h1> Plasmids containing toxin ccdB</h1> | + | |
| - | <h2>pSB3T5 T7 ccdB</h2> | + | <center><img src="https://static.igem.org/mediawiki/2013/e/e9/UGent_2013_PTGD-ccdA-Pmb1-BCD7-GFP-CmFRT.png" width="700"/></center> |
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| + | <h1> Plasmids containing toxin <i> ccdB </i></h1> | ||
| + | <p> All of these plasmids are constructed using BioBrick standard parts: the plasmid backbones pSB3T5, pSB4A5 and pSB6A1. The <i>ccdB</i> gene that encodes the toxin is controlled by a T7 promoter. </p> | ||
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| + | <h2>pSB3T5-T7<i>ccdB </i></h2> | ||
<center><img src="https://static.igem.org/mediawiki/2013/4/4a/UGent_2013_PSB3T5-T7-ccdB.png" width="500"/></center> | <center><img src="https://static.igem.org/mediawiki/2013/4/4a/UGent_2013_PSB3T5-T7-ccdB.png" width="500"/></center> | ||
| - | <h2>pSB4A5 T7 ccdB</h2> | + | <h2>pSB4A5-T7<i>ccdB </i></h2> |
<center><img src="https://static.igem.org/mediawiki/2013/b/bc/UGent_2013_PSB4A5-T7-ccdB.png" width="500"/></center> | <center><img src="https://static.igem.org/mediawiki/2013/b/bc/UGent_2013_PSB4A5-T7-ccdB.png" width="500"/></center> | ||
| - | <h2>pSB6A1 T7 ccdB</h2> | + | <h2>pSB6A1-T7<i>ccdB </i></h2> |
<center><img src="https://static.igem.org/mediawiki/2013/b/b1/UGent_2013_PSB6A1-T7-ccdB.png" width="500"/></center> | <center><img src="https://static.igem.org/mediawiki/2013/b/b1/UGent_2013_PSB6A1-T7-ccdB.png" width="500"/></center> | ||
| - | <h1> | + | <h1> Plasmid containing new part </h1> |
| - | <h2>pSB1C3 T7 ccdB</h2> | + | <h2>pSB1C3-T7<i>ccdB </i></h2> |
<center><img src="https://static.igem.org/mediawiki/2013/c/c1/UGent_2013_PSB1C3-T7-ccdB.png" width="600"/></center> | <center><img src="https://static.igem.org/mediawiki/2013/c/c1/UGent_2013_PSB1C3-T7-ccdB.png" width="600"/></center> | ||
Latest revision as of 01:50, 5 October 2013
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To perform in silico analysis, we used Clone Manager. All plasmids were first implemented into the software, and then used to help with cloning simulation, restriction, PCR etc. Plasmid containing construct for chromosomal evolutionpTGD ccdA Pmb1 BCD7 GFP CmFRTThis plasmid contains the ccdA antitoxin and the reporter gene gfp. The homologous regions are necessary to perform Chemically Inducible Chromosomal evolution (CIChE). If wanted, the removal of the chloramphenicol resistance cassette between the FTR sites can be done using the method of Datsenko & Wanner [PNAS 2000].  Plasmids containing toxin ccdBAll of these plasmids are constructed using BioBrick standard parts: the plasmid backbones pSB3T5, pSB4A5 and pSB6A1. The ccdB gene that encodes the toxin is controlled by a T7 promoter. pSB3T5-T7ccdB pSB4A5-T7ccdB pSB6A1-T7ccdB Plasmid containing new partpSB1C3-T7ccdB
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