http://2014.igem.org/wiki/index.php?title=Special:Contributions/Andy.Bachler&feed=atom&limit=50&target=Andy.Bachler&year=&month=2014.igem.org - User contributions [en]2024-03-28T15:05:46ZFrom 2014.igem.orgMediaWiki 1.16.5http://2014.igem.org/Template:Team:USyd-Australia/Template:StyleTemplate:Team:USyd-Australia/Template:Style2014-10-18T03:54:38Z<p>Andy.Bachler: </p>
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</html></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T03:54:21Z<p>Andy.Bachler: </p>
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<div>{{Team:USyd-Australia/Template:Style}}<br />
{{:Team:USyd-Australia/Template:Banner}}<br />
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<html><br />
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<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template.</p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table id="gBlockTable"><br />
<br />
<tr id="gBlockTableRow"><th id="gBlockTableHeading" width="10%">Name</th><th id="gBlockTableHeading" width="45%">Sequence (5'-3')</th><th id="gBlockTableHeading" width="5%">Size</th><th id="gBlockTableHeading" width="20%">Purpose</th></tr><br />
<br />
<tr id="gBlockTableRow"><td id="gBlockTableColumn"><a name="IntI1.2"></a>IntI1</td><td id="gBlockTableColumn" style="word-wrap: break-word; font-size: 8px;">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td id="gBlockTableColumn">1121 bp</td><td id="gBlockTableColumn">To be inserted into SamR construct as part of the controllable integrase expression system</td></tr><br />
<br />
<tr id="gBlockTableRow"><td id="gBlockTableColumn"><a name="AttC-aeBlue-aacC1 "></a>AttC-aeBlue-aacC1 </td><td id="gBlockTableColumn" style="word-wrap: break-word; font-size: 8px;">TTTGAATTCGCGGCCGCTTCTAGAGTACCGAGGGACAGACTACCAACTCACAGCTAGCGCCTAACAATTCGTCCAAGCCGACGCCGCTTCGCGGCGCGGCTTAACTCAGGTGTTAGGCAAAGGAGGTTATTGATATGGCTTCACTGGTTAAAAAAGACATGTGCATCAAAATGACGATGGAAGGAACAGTAAACGGTCACCATTTCAAGTGTGTAGGAGAAGGCGAAGGCAAACCATTTGAAGGGACCCAGGTGGAAAAGATACGCATCACTGAAGGTGGGCCCTTACCATTTGCGTATGATATTTTGGCCCCTTGTTGCATGTATGGCAGTAAAACCTTCATTAAGCATGTGTCGGGTATTCCGGATTACTTTAAGGAGTCTTTTCCTGAGGGCTTTACCTGGGAAAGAACACAAATCTTCGAGGATGGCGGCTATCTCACCATACACCAGGACACGAGCCTTCAGGGTAATAATTTTATTTTCAAAGTTAATGTCATCGGTGCCAACTTCCCTGCAAACGGTCCCGTGATGCAGAAAAAAACAGCTGGATGGGAACCGTGCGTTGAGATGCTTTATCCGCGGGACGGCGTCCTGTGTGGTCAGAGCCTGATGGCCCTGAAATGCACTGATGGCAATCATCTGACGTCCCACCTGCGCACTACCTATCGTTCTCGCAAGCCATCCAATGCAGTTAACATGCCGGAATTTCATTTTGGGGATCATCGCATTGAGATTTTGAAAGCTGAACAAGGTAAATTTTATGAACAATACGAGTCAGCGGTGGCCCGTTACTGTGAGGCGGCACCGAGTAAATTAGGGCATCACTAATAAAGGATCCAAAGGAGGCTCAAGTATGGGCATCATTCGCACATGTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGATCTATATCTATGATCTCGCAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCGCGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGGAAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAATAATACTAGTAGCGGCCGCTGCAGCCC</td><td id="gBlockTableColumn">1347 bp</td><td id="gBlockTableColumn">To be used as a template for the creation of gene cassettes with antibiotic resistance and phenotypic markers</td></tr><br />
<br />
<tr id="gBlockTableRow"><td id="gBlockTableColumn"><a name="LacI-lacO-pLac-attI"></a>LacI-lacO-pLac-attI </td><td id="gBlockTableColumn" style="word-wrap: break-word; font-size: 8px;">GGGAATTCAAATCTAGAGACACCATCGAATGGTGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAATGAAGGAGGTGAATATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTCAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTAATAAGCGCAAAAAAAAAACCCCGCCCCTGACAGGGCGGGGTTTTTTTTCAACGCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAGGCAAAACTAGTAAACTGCAGGG</td><td id="gBlockTableColumn">1403 bp</td><td id="gBlockTableColumn">To be used as an AttI site with a Lac operon to allow controllable expression</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T03:53:43Z<p>Andy.Bachler: </p>
<hr />
<div>{{Team:USyd-Australia/Template:Style}}<br />
{{:Team:USyd-Australia/Template:Banner}}<br />
<br />
<html><br />
<br />
<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template.</p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table id="gBlockTable"><br />
<br />
<tr id="gBlockTableRow"><th id="gBlockTableRow" width="10%">Name</th><th id="gBlockTableRow" width="45%">Sequence (5'-3')</th><th id="gBlockTableRow" width="5%">Size</th><th id="gBlockTableRow" width="20%">Purpose</th></tr><br />
<br />
<tr id="gBlockTableRow"><td id="gBlockTableColumn"><a name="IntI1.2"></a>IntI1</td><td id="gBlockTableColumn" style="word-wrap: break-word; font-size: 8px;">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td id="gBlockTableColumn">1121 bp</td><td id="gBlockTableColumn">To be inserted into SamR construct as part of the controllable integrase expression system</td></tr><br />
<br />
<tr id="gBlockTableRow"><td id="gBlockTableColumn"><a name="AttC-aeBlue-aacC1 "></a>AttC-aeBlue-aacC1 </td><td id="gBlockTableColumn" style="word-wrap: break-word; font-size: 8px;">TTTGAATTCGCGGCCGCTTCTAGAGTACCGAGGGACAGACTACCAACTCACAGCTAGCGCCTAACAATTCGTCCAAGCCGACGCCGCTTCGCGGCGCGGCTTAACTCAGGTGTTAGGCAAAGGAGGTTATTGATATGGCTTCACTGGTTAAAAAAGACATGTGCATCAAAATGACGATGGAAGGAACAGTAAACGGTCACCATTTCAAGTGTGTAGGAGAAGGCGAAGGCAAACCATTTGAAGGGACCCAGGTGGAAAAGATACGCATCACTGAAGGTGGGCCCTTACCATTTGCGTATGATATTTTGGCCCCTTGTTGCATGTATGGCAGTAAAACCTTCATTAAGCATGTGTCGGGTATTCCGGATTACTTTAAGGAGTCTTTTCCTGAGGGCTTTACCTGGGAAAGAACACAAATCTTCGAGGATGGCGGCTATCTCACCATACACCAGGACACGAGCCTTCAGGGTAATAATTTTATTTTCAAAGTTAATGTCATCGGTGCCAACTTCCCTGCAAACGGTCCCGTGATGCAGAAAAAAACAGCTGGATGGGAACCGTGCGTTGAGATGCTTTATCCGCGGGACGGCGTCCTGTGTGGTCAGAGCCTGATGGCCCTGAAATGCACTGATGGCAATCATCTGACGTCCCACCTGCGCACTACCTATCGTTCTCGCAAGCCATCCAATGCAGTTAACATGCCGGAATTTCATTTTGGGGATCATCGCATTGAGATTTTGAAAGCTGAACAAGGTAAATTTTATGAACAATACGAGTCAGCGGTGGCCCGTTACTGTGAGGCGGCACCGAGTAAATTAGGGCATCACTAATAAAGGATCCAAAGGAGGCTCAAGTATGGGCATCATTCGCACATGTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGATCTATATCTATGATCTCGCAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCGCGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGGAAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAATAATACTAGTAGCGGCCGCTGCAGCCC</td><td id="gBlockTableColumn">1347 bp</td><td id="gBlockTableColumn">To be used as a template for the creation of gene cassettes with antibiotic resistance and phenotypic markers</td></tr><br />
<br />
<tr id="gBlockTableRow"><td id="gBlockTableColumn"><a name="LacI-lacO-pLac-attI"></a>LacI-lacO-pLac-attI </td><td id="gBlockTableColumn" style="word-wrap: break-word; font-size: 8px;">GGGAATTCAAATCTAGAGACACCATCGAATGGTGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAATGAAGGAGGTGAATATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTCAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTAATAAGCGCAAAAAAAAAACCCCGCCCCTGACAGGGCGGGGTTTTTTTTCAACGCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAGGCAAAACTAGTAAACTGCAGGG</td><td id="gBlockTableColumn">1403 bp</td><td id="gBlockTableColumn">To be used as an AttI site with a Lac operon to allow controllable expression</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Template:Team:USyd-Australia/Template:StyleTemplate:Team:USyd-Australia/Template:Style2014-10-18T03:51:09Z<p>Andy.Bachler: </p>
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</html></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Template:BannerTeam:USyd-Australia/Template:Banner2014-10-18T03:44:16Z<p>Andy.Bachler: </p>
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<br />
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<br />
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</tr><br />
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<a href="https://2014.igem.org/Team:USyd-Australia/Team"><br />
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<a href="https://2014.igem.org/Team:USyd-Australia/Project/Design"><br />
<img src="https://static.igem.org/mediawiki/2014/a/a3/USyd-Australia_Integron_basic_diagram.png" width="390px"><br />
</a></td><br />
<br />
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<a href="https://2014.igem.org/Team:USyd-Australia/Outreach"><br />
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</a></td><br />
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<tr><td colspan="5 align="center"><h3>Integrons as a novel cloning system in E. coli</h3></td></tr><br />
<br />
<tr><br />
<td colspan="5" align="justify"><a href="https://2014.igem.org/Team:USyd-Australia/Project/Background">Integrons</a> are natural systems for moving genetic material. They are found in a vast array of bacteria, often associated with rapidly evolving traits such as antibiotic resistance. This is because integrons capture and express mobile genetic elements known as gene cassettes, thus facilitating horizontal gene transfer. Gene cassettes are site selectively recombined by the integron integrase. We have <a href="https://2014.igem.org/Team:USyd-Australia/Project/Design">designed</a> BioBrick- compatible integron components that can be used as a novel, convenient and selective cloning system. Additionally, we have been investigating regulation of natural transformation mechanisms in E. coli in an attempt to allow uptake and recombination of attC containing genetic elements with added convenience. </td><br />
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{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-AustraliaTeam:USyd-Australia2014-10-18T03:40:35Z<p>Andy.Bachler: </p>
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<br />
<td width="30%" align="center"><h3>Strange Nature</h3></td><br />
</tr><br />
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<a href="https://2014.igem.org/Team:USyd-Australia/Team"><br />
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<img src="https://static.igem.org/mediawiki/2014/e/e1/USyd-Australia_Profile-Jeanne.jpg" width="125px" vspace="2px"> <br><br />
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</a><br />
<br />
<td align="center"><br />
<a href="https://2014.igem.org/Team:USyd-Australia/Project/Design"><br />
<img src="https://static.igem.org/mediawiki/2014/a/a3/USyd-Australia_Integron_basic_diagram.png" width="390px"><br />
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<a href="https://2014.igem.org/Team:USyd-Australia/Outreach"><br />
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</a></td><br />
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<br />
<br />
<tr><td colspan="5 align="center"><h3>Integrons as a novel cloning system in E. coli</h3></td></tr><br />
<br />
<tr><br />
<td colspan="5" align="justify"><a href="https://2014.igem.org/Team:USyd-Australia/Project/Background">Integrons</a> are natural systems for moving genetic material. They are found in a vast array of bacteria, often associated with rapidly evolving traits such as antibiotic resistance. This is because integrons capture and express mobile genetic elements known as gene cassettes, thus facilitating horizontal gene transfer. Gene cassettes are site selectively recombined by the integron integrase. We have <a href="https://2014.igem.org/Team:USyd-Australia/Project/Design">designed</a> BioBrick- compatible integron components that can be used as a novel, convenient and selective cloning system. Additionally, we have been investigating regulation of natural transformation mechanisms in E. coli in an attempt to allow uptake and recombination of attC containing genetic elements with added convenience. </td><br />
<br />
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<br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Template:Team:USyd-Australia/Template:StyleTemplate:Team:USyd-Australia/Template:Style2014-10-18T03:40:04Z<p>Andy.Bachler: </p>
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</html></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Project/BackgroundTeam:USyd-Australia/Project/Background2014-10-18T03:36:38Z<p>Andy.Bachler: </p>
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<h2>Project Background</h2><br />
<br />
<br />
<h3>What are integrons?</h3><br />
<br />
<p>Integrons are systems which can capture and utilise mobile genetic elements known as gene cassettes, and which are found in a vast array of bacteria. Cassettes are often associated with antibiotic resistance and are a significant factor in the spread of antibiotic resistance between bacteria. Gene cassettes with an attC recognition site are site selectively recombined into an attI recognition site by the integron integrase. Insertion of cassettes can continue to occur at the AttI site, and with much lower frequency at the AttC sites, resulting in a long array of cassettes which can be expressed from a constitutive promoter (Pc) just upstream from the AttI. Cassettes may also be <a href="http://www.ncbi.nlm.nih.gov/pubmed/1331702"> from the array</a>, </p><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/c/c4/USyd-Australia_Integrons-Class1.png" width="300px" align="right"><br />
<br />
<p>There are a few classes of integrons, but the first to be discovered and most well-known are the Class 1 integrons. Class 1 integrons are the most medically important class because of their involvement with antibiotic resistance in pathogenic bacteria. As shown in the diagram on the right, Class 1 integrons are defined by the presence of a conserved sulfonamide resistance gene on the outside of the array. The cassettes found within class 1 integrons are usually antibiotic resistance genes.</p><br />
<br />
<p>Since the integration of new cassettes is simultaneous with expression by the constitutive promoter, the system can conceivably be used for pathway optimisation by <a href="http://www.ncbi.nlm.nih.gov/pubmed/20534632"> gene shuffling</a>. This means that new insertions are immediately tested and selected for if beneficial. <br />
</p><br />
<br />
<p>An extensive review about integrons can be found <a href="http://www.nature.com/nrmicro/journal/v4/n8/pdf/nrmicro1462.pdf">here</a>.<br />
<br />
<h3>What are we doing?</h3><br />
<br />
<p>We are seeking to design biobrick compatible, completely controllable class 1 integron components that can be used as a novel, convenient and selective cloning system in lab strains of E. coli, which normally do not contain integron systems. Additionally, we are investigating regulation of natural transformation mechanisms in E. coli in an attempt to allow uptake and recombination of attC containing genetic elements with added convenience. </p><br />
<br />
<h3>Why is this cool?</h3><br />
<br />
<p>By re-building nature’s modular genetic shuffling machine, we aim to provide a novel tool for Synthetic Biologists AND ALSO create an experimental system in which we can more precisely understand integrons.</p><br />
<br />
<h3>Integrons in iGEM</h3><br />
<br />
<p>Integrons have been encountered before in iGEM for different purposes:<br />
<ul><br />
<li>A molecular ‘counter’,<br />
<a href="https://2010.igem.org/Team:Paris_Liliane_Bettencourt/Project/Population_counter/Design">Paris Liliane Bettencourt, 2010</a><br />
<li>Gene shuffling for pathway optimisation,<br />
<a href="https://2012.igem.org/Team:ULB-Brussels">ULB Brussels, 2012</a><br />
</ul><br />
<h3>Further Reading</h3><br />
<p> <a href="http://www.nature.com/nrmicro/journal/v4/n8/abs/nrmicro1462.html">Nature Review Article</a> talking about Integrons and bacterial evolution.</p><br />
<p> <a href="https://static.igem.org/mediawiki/2014/a/ac/Hall%2BCollis-integron-review-MolMic1995.pdf">Earlier Review</a> by University of Sydney Professor Ruth Hall.<br />
<p> The <a href="https://static.igem.org/mediawiki/2014/2/2d/Guerin-Mazel-SOS-response-integrons-Science2009.pdf">SOS Response and Integrons</a><br />
<br />
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{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/File:Guerin-Mazel-SOS-response-integrons-Science2009.pdfFile:Guerin-Mazel-SOS-response-integrons-Science2009.pdf2014-10-18T03:36:10Z<p>Andy.Bachler: </p>
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<div></div>Andy.Bachlerhttp://2014.igem.org/File:Hall%2BCollis-integron-review-MolMic1995.pdfFile:Hall+Collis-integron-review-MolMic1995.pdf2014-10-18T03:33:49Z<p>Andy.Bachler: </p>
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<div></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/AttributionsTeam:USyd-Australia/Attributions2014-10-18T03:29:10Z<p>Andy.Bachler: </p>
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<h1> Attributions </h1><br />
<p> <b> Dr Nick Coleman</b> - Our brilliant supervisor, providing constant support and guidance no matter how many times we forgot to run controls. </p><br />
<p> <b> Robbie Oppenheimer </b> - The founder of iGEM at Sydney University who has been a tremendous advisor and motivator throughout the whole year. </p><br />
<p> <b> The <a href = https://2013.igem.org/Team:SydneyUni_Australia > 2013 USYD iGEM team!</a> </b> </p><br />
<p> <b> Dr Hannah Nicholas </b>- For providing guidance at the beginning of our project. </p><br />
<p> <b> Elissa Liew, Dr Mai-Anh Ly, Sam Ross and numerous others </b>- Who all helped our team navigate the lab and pointed out the mistakes we were making in real time! </p><br><br />
<br />
<p> Thanks again to <b>Sam Ross</b>, for providing the AraC-pBad controllable promoter system in pSB1C3. </p><br />
<br />
<h1> Financial and Material Support </h1><br />
<br />
<p> Thank you for support from: </p><br />
<p> <b> Prof. Trevor Hambley </b> - Dean of the Faculty of Science, University of Sydney. </p><br />
<p> <b> Prof. Ian Campbell </b> - Head of the School of Molecular Bioscience, University of Sydney. </p><br />
<br><br />
<br />
<p> <b> The <a href = http://www.snapgene.com/ > Snapgene </a> team </b> - For allowing us to use Snapgene for free, it was an invaluable tool for planning our project. </p><br />
<p> <b> Dr Paul Priscott </b> - <a href="http://www.amslabs.com.au/"> AMS Laboratories</a>, for prize money for our strange nature writing competition.</p><br><br />
<br />
<h1> Strange Nature judges </h1><br />
<p> Thanks to the judges of our <a href="http://strangenature.org/"> Strange Nature Writing competition</a>:</p><br />
<p> <b> Yagiz Aksoy, </b> from Macquarie University </p><br />
<p> <b> Jasmine Fellows, </b> editor of CSIRO Helix Magazine</p><br />
<p> <b> Jane McCredie, </b> from NSW Writers' Centre </p><br />
<p> <b> Dr Claudia Vickers, </b> from the University of Queensland </p><br />
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{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Template:Team:USyd-Australia/Template:StyleTemplate:Team:USyd-Australia/Template:Style2014-10-18T03:26:16Z<p>Andy.Bachler: </p>
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</html></div>Andy.Bachlerhttp://2014.igem.org/Template:Team:USyd-Australia/Template:StyleTemplate:Team:USyd-Australia/Template:Style2014-10-18T03:24:15Z<p>Andy.Bachler: </p>
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</html></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Project/ResultsConclusionTeam:USyd-Australia/Project/ResultsConclusion2014-10-18T03:09:10Z<p>Andy.Bachler: </p>
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<h2>Results of Components</h2><br />
<br />
<h3>pUS201 - Low Copy Backbone</h3><br />
<p>Characterization of the low-copy backbone was unfortunately unsuccessful. Despite careful planning and verification of correct insertion and sequence the low-copy backbone displayed high copy replication characteristics. See the <a href="https://2014.igem.org/Team:USyd-Australia/pUS201">pUS201 Parts page</a> for more info. <br />
<br />
<h3>pUS203 - Controllable Integrase</h3><br />
<p>Without pUS201 the experimental design had to adapt. The controllable Integrase system from pUS203 should ideally be in a low copy backbone but the the iGEM supplied low-copy plasmid was not an option as the anti-biotic resistance interfered with the experimental set up. At this point in time we did not have time to alter the experiment to such a degree as to accommodate the change. Instead we set about validating the controllable integrase system in the submission plasmid to try and determine whether it was producing integrase but the initial experiment failed and follow up experiments to troubleshoot problems were not possible due to time constraints. See the <a href="https://2014.igem.org/Team:USyd-Australia/pUS203">pUS203 Parts page</a> for more info.<br />
<br />
<h3>pUS204 - Cassette in pSB1C3</h3> The cassette was functionally proven although it did not express the aeBlue gene but did confer gentamycin resistance. After sequencing we have found that a deletion near the end of the part seems to have been the cause for the absence of blue from the colonies expressing the cassette. Despite the lack of blue reporter phenotype being observed, the Gm Resistance was real. This part can be considered as ready to use for further experiments involving integrons.</p><br />
<br />
<h2>Conclusion of Project</h2><br />
<br />
<p>At the time of the Wiki Freeze we have been unable to functionally validate our controllable integrase BioBrick and so cannot determine if it expressing Integrase under the action of arabinose or not. Troubleshooting of the original experiment and designing a new experiment to allow us to use the low-copy plasmid would give us evidence for which part of the araC-pBAD system isn't working but due to time constraints we could not perform these tests before the end of the competition. <br><br>The low-copy plasmid has been shown to be high-copy and so we believed it was not worth submitting as a part as it was functionally incorrect.<br><br> The cassette worked but with only the Gentamicin resistance as a selectable marker. We view this as a great success as it's a definite step toward creation of our Integrase cloning system if another iGEM team or next years USyd team could validate a controllable Integrase BioBrick!</p><br />
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</html></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/pUS203Team:USyd-Australia/pUS2032014-10-18T02:55:49Z<p>Andy.Bachler: </p>
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<a name="protstart"></a><br />
<h2><a href="http://parts.igem.org/Part:BBa_K1388000">pUS203 Controllable Integrase production</a></h2><br />
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<p style="font-size:12px"><br />
To see this part on the iGEM Parts catalogue please click <a href="http://parts.igem.org/Part:BBa_K1388000">here</a>. <br>Sequences for this part are also available in <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLRDJ3OVdUUlhEVjA/view?usp=sharing">SnapGene (.dna file)</a>, <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLT3hQZno4VXFVZ0U/view?usp=sharing">FASTA (.fa file)</a>, <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLbndkclF5UlFNVE0/view?usp=sharing">GenBank Standard (.gb file)</a> and <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLak5Qdi1CbFZ4bm8/view?usp=sharing">Plain Text (.txt file)</a> file format.<br />
</p><br />
<h2>Aim</h2><br />
<p>Our aim was to create a controllable Integrase producing plasmid as a submission part. From the submission plasmid the controllable Integrase Part could then be ligated into a convenient low-copy plasmid to help reduce the potential toxicity of Integrase to the cell.</p><br />
<br />
<h2>Approach</h2><br />
<p><br />
<img src="https://static.igem.org/mediawiki/2014/6/67/PUS203.png" align="right" width="50%"><br />
The 2010 Paris Bettencourt team had developed a <a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K329013>controllable Integrase part</a> for use a population counter in their project. We contacted them to see if we could source the part from them but it was unfortunately not possible. We set about creating the BioBrick using the araC-pBAD from the well characterized part <a href=http://parts.igem.org/Part:BBa_K731201>K731201</a> and ordering the IntI1 as a gBlock. The araC-pBAD system in pSB1C3 had previously been prepared by Honours student Sam Ross (called the SamR construct). Our next steps were the design of the gBlock and integration of the IntI1 gBlock into the SamR construct.<br />
</p><br />
<br />
<h2>Materials</h2><br />
<ul>DNA<ul><br />
<li>SamR Construct: araC-pBAD in pSC1B3</li><br />
<li><a href="#">IntI1 gBlock</a>: to be inserted into SamR</li><br />
<li>pUS44: Verified plasmid containing an AttI site, used in Validation</li><br />
<li>aeBlue: Cassette PCR'ed from <a href=http://parts.igem.org/Part:BBa_K1033929>K1033929</a> for us in validation</li><br />
</ul><br />
</ul><br />
<br />
<ul>Primers<br />
<table><br />
<tr><br />
<td width="50%"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1413">iGEM1413</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1414">iGEM1414</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1415">iGEM1415</a></li><br />
</ul><br />
</td><br />
<td><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1416">iGEM1416</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers">iGEM16</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers">iGEM25</a></li><br />
</ul><br />
</td><br />
</tr><br />
<tr><br />
</table><br />
<br />
<h2>Inti1 gBlock Design</h2><br />
<p><br />
The code for the sequence for the IntI1 gene was sourced from pUS2056 but is highly conserved so this choice was out of convenience as the sequence was locally available. It was modified by Tom Geddes, Rokiah Alford and our supervisor Dr Nicholas Coleman to:<br><ul><br />
<li>Include a Sac1 restriction enzyme site at the beginning before the Inti1 gene so it could be excised independently of the promoter if needed</li><br />
<li>Include a different ribosome binding site </li><br />
<li>Include Gibson ends that overlapped with the backbone for simple Gibson Assembly later with the SamR construct.</li><br />
<li>Remove an illegal Spe1 site near the end to make it compliant with iGEM requirements</li><br />
<li>Remove a promoter pC that was coded in the opposite direction to the promoter we wanted to use and would have clashed with the controllable promotion of this part</li></ul><br><br />
Our first attempt to Gibson Assemble the SamR construct and IntI1 gBlock failed because there was extra code after the code for pBAD which we had not anticipated. This meant that the Gibson Assembly could not properly integrate the gBlock. We discussed numerous options to try and either get rid of the problem sequence on the SamR construct or modify the ends of the gBlock to accommodate the problem sequence. In the end it was decided that a re-design of the gBlock would be the most efficient way to solve the problem. <br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<h2>Inti1 gBlock ReDesign</h2><br />
<p><br />
Three major changes were decided: <br><br />
<ul><br />
<li>Changed the code for the ribosome cut site as a base pair was incorrect</li><br />
<li>To put an Nhe1 restriction enzyme cut site between the IntI1 gene and the promoters on the SamR construct. This was done to help in the ligation of this BioBrick into the low-copy backbone pUS202. </li><br />
<li>Changing of the end sequences to accommodate the extra code present on the SamR construct</li><br />
</ul><br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<br />
<h2>Construction</h2><br />
<p><br />
The IntI1 gBlock was inserted into the SamR construct using <a href="http://www.addgene.org/plasmid-protocols/gibson-assembly/">Gibson Asssembly</a>. The ends of the gBlock in the redesign were compatible with the ends of the SamR construct and after running the Gibson Assembly reaction we used a variety of methods to prove that our gBlock had inserted correctly.<br />
</p><br />
<br />
<p><br />
These changed were performed by our supervisor Nick Coleman in consultation with Rokiah Alford. The redesigned gBlock was able to be successfully Gibsoned into the SamR backbone. To verify the presence of the insert we used junction primers which would amplify over both ends of the insert and could only amplify if the gBlock had been integrated into the SamR construct. <br><br />
<br><b>Junction primer PCR</b><br><br />
<br />
Junction primers for the junction of promoter and gblock were iGEM1413 and iGEM1414 and were expected to give a fragment size of 377 base pairs. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/4/4e/USyd-AustraliaWell_Images_restriction_Digest.png"> <br><br />
<br />
9 out of the 12 colonies screened gave a positive result and these were further screened with a second set of junction primers on the other side of the gBlock. The junction primers for this were iGEM1416 and iGEM25 and were expected to yielad a fragment size of 432bp. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/7/70/USyd-Australia_RealSecondPCR_Junction_screen.png"> <br><br />
<br />
Again 9 of the 12 colonies were positive here. The PCR product was sequenced and corresponded with the expected sequence for the plasmid. 3 of the 8 colonies were selected to be grown up and Plasmid Prepped to remove the plasmid and allow restriction digest and sequencing of the plasmid for further verification.<br><br />
<br />
<br><b>Restriction Digest</b><br><br />
<br />
The pUS203 plasmid was further confirmed with restriction digestion. Spe1, Pst1 and EcoR1 are in the prefix and suffix and should cut. An Nhe1 restriction site was added into the gBlock code and so a cut with this enzyme would indicate the gBlock had been inserted. Sac1 should not cut and should be identical to the uncut control lane. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/4/4e/USyd-AustraliaWell_Images_restriction_Digest.png"><br><br />
<br />
The digest results were as expected using SnapGene’s Virtual Digest tool. We were satisfied with these results despite the Sac1 error because the other fragments were as expected and most importantly because Nhe1 cut, which could only have occurred if the gblock had been incorporated into the plasmid. The Sac1 sample was heat-treated to inactive the Sac1 enzyme and used was used as a template to PCR the BioBrick. The results were still as expected which showed that the unexpected action of the Sac1 seen in the gel for the digest of this plasmid luckily didn’t reflect an error in the biobrick code. <br><br><br />
<br />
<b>Biobrick PCR and sequencing</b><br><br />
The final check for this plasmid was to PCR the entire biobrick fragment using iGEM16 and iGEM25 and sequencing the product. <br />
</p><br />
<br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<h2>Validation</h2><br />
<ul style="font-weight: normal;"><br />
To validate our araC-pBAD/IntI1 BioBrick it would need to be present in a low copy plasmid in order to reduce the amount produced as it is potentially toxic to the cell at high concentrations. Our intial plan was to insert the BioBrick into pUS202 but it turned out not to be low-copy after validation. The back up option was to ligate the BioBrick into pSB6A1. The initial attempt was not successful and due to time constraints we decided to try and test the integron construct in the pSB1C3 it was currently in even with the potential toxicity.<br><br><br />
To test for the proper expression of our integron gene in the BioBrick we wanted to have an AttI site and integrase protein present in E. coli. This was so that upon transformation with a cassette (which has an AttC site) a co-integrate would quickly form with the AttI site and before the cassettes could be lost out of the cell. Upon integration of the cassette the Gentomycin resistance (GmR) present on the cassette is expressed allowing us to select for the correct cells on Gentomycin plates. The negative control had no arabinose induction and so the cassette should not integrate and the cells should not be Gm resistant.<br><br><br />
<img src="https://static.igem.org/mediawiki/2014/7/71/USyd-Australia_Integrase_diagrams_1.png" width="98%"><br><br><br />
The initial experiments did not yield positive results and due to time constraints we have not been able to troubleshoot the experiment. <br />
</ul><br />
</li><br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
</html></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/pUS203Team:USyd-Australia/pUS2032014-10-18T02:55:21Z<p>Andy.Bachler: </p>
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<h2><a href="http://parts.igem.org/Part:BBa_K1388000">pUS203 Controllable Integrase production</a></h2><br />
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<p style="font-size:12px"><br />
To see this part on the iGEM Parts catalogue please click <a href="http://parts.igem.org/Part:BBa_K1388000">here</a>. <br>Sequences for this part are also available in <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLRDJ3OVdUUlhEVjA/view?usp=sharing">SnapGene (.dna file)</a>, <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLT3hQZno4VXFVZ0U/view?usp=sharing">FASTA (.fa file)</a>, <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLbndkclF5UlFNVE0/view?usp=sharing">GenBank Standard (.gb file)</a> and <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLak5Qdi1CbFZ4bm8/view?usp=sharing">Plain Text (.txt file)</a> file format.<br />
</p><br />
<h2>Aim</h2><br />
<p>Our aim was to create a controllable Integrase producing plasmid as a submission part. From the submission plasmid the controllable Integrase Part could then be ligated into a convenient low-copy plasmid to help reduce the potential toxicity of Integrase to the cell.</p><br />
<br />
<h2>Approach</h2><br />
<p><br />
<img src="https://static.igem.org/mediawiki/2014/6/67/PUS203.png" align="right" width="50%"><br />
The 2010 Paris Bettencourt team had developed a <a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K329013>controllable Integrase part</a> for use a population counter in their project. We contacted them to see if we could source the part from them but it was unfortunately not possible. We set about creating the BioBrick using the araC-pBAD from the well characterized part <a href=http://parts.igem.org/Part:BBa_K731201>K731201</a> and ordering the IntI1 as a gBlock. The araC-pBAD system in pSB1C3 had previously been prepared by Honours student Sam Ross (called the SamR construct). Our next steps were the design of the gBlock and integration of the IntI1 gBlock into the SamR construct.<br />
</p><br />
<br />
<h2>Materials</h2><br />
<ul>DNA<ul><br />
<li>SamR Construct: araC-pBAD in pSC1B3</li><br />
<li><a href="#">IntI1 gBlock</a>: to be inserted into SamR</li><br />
<li>pUS44: Verified plasmid containing an AttI site, used in Validation</li><br />
<li>aeBlue: Cassette PCR'ed from <a href=http://parts.igem.org/Part:BBa_K1033929>K1033929</a> for us in validation</li><br />
</ul><br />
</ul><br />
<br />
<ul>Primers<br />
<table><br />
<tr><br />
<td width="50%"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1413">iGEM1413</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1414">iGEM1414</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1415">iGEM1415</a></li><br />
</ul><br />
</td><br />
<td><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1416">iGEM1416</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers">iGEM16</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers">iGEM25</a></li><br />
</ul><br />
</td><br />
</tr><br />
<tr><br />
</table><br />
<br />
<h2>Inti1 gBlock Design</h2><br />
<p><br />
The code for the sequence for the IntI1 gene was sourced from pUS2056 but is highly conserved so this choice was out of convenience as the sequence was locally available. It was modified by Tom Geddes, Rokiah Alford and our supervisor Dr Nicholas Coleman to:<br><ul><br />
<li>Include a Sac1 restriction enzyme site at the beginning before the Inti1 gene so it could be excised independently of the promoter if needed</li><br />
<li>Include a different ribosome binding site </li><br />
<li>Include Gibson ends that overlapped with the backbone for simple Gibson Assembly later with the SamR construct.</li><br />
<li>Remove an illegal Spe1 site near the end to make it compliant with iGEM requirements</li><br />
<li>Remove a promoter pC that was coded in the opposite direction to the promoter we wanted to use and would have clashed with the controllable promotion of this part</li></ul><br><br />
Our first attempt to Gibson Assemble the SamR construct and IntI1 gBlock failed because there was extra code after the code for pBAD which we had not anticipated. This meant that the Gibson Assembly could not properly integrate the gBlock. We discussed numerous options to try and either get rid of the problem sequence on the SamR construct or modify the ends of the gBlock to accommodate the problem sequence. In the end it was decided that a re-design of the gBlock would be the most efficient way to solve the problem. <br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<h2>Inti1 gBlock ReDesign</h2><br />
<p><br />
Three major changes were decided: <br><br />
<ul><br />
<li>Changed the code for the ribosome cut site as a base pair was incorrect</li><br />
<li>To put an Nhe1 restriction enzyme cut site between the IntI1 gene and the promoters on the SamR construct. This was done to help in the ligation of this BioBrick into the low-copy backbone pUS202. </li><br />
<li>Changing of the end sequences to accommodate the extra code present on the SamR construct</li><br />
</ul><br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<br />
<h2>Construction</h2><br />
<p><br />
The IntI1 gBlock was inserted into the SamR construct using <a href="http://www.addgene.org/plasmid-protocols/gibson-assembly/">Gibson Asssembly</a>. The ends of the gBlock in the redesign were compatible with the ends of the SamR construct and after running the Gibson Assembly reaction we used a variety of methods to prove that our gBlock had inserted correctly.<br />
</p><br />
<br />
<p><br />
These changed were performed by our supervisor Nick Coleman in consultation with Rokiah Alford. The redesigned gBlock was able to be successfully Gibsoned into the SamR backbone. To verify the presence of the insert we used junction primers which would amplify over both ends of the insert and could only amplify if the gBlock had been integrated into the SamR construct. <br><br />
<br><b>Junction primer PCR</b><br><br />
<br />
Junction primers for the junction of promoter and gblock were iGEM1413 and iGEM1414 and were expected to give a fragment size of 377 base pairs. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/4/4e/USyd-AustraliaWell_Images_restriction_Digest.png"> <br><br />
<br />
9 out of the 12 colonies screened gave a positive result and these were further screened with a second set of junction primers on the other side of the gBlock. The junction primers for this were iGEM1416 and iGEM25 and were expected to yielad a fragment size of 432bp. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/7/70/USyd-Australia_RealSecondPCR_Junction_screen.png"> <br><br />
<br />
Again 9 of the 12 colonies were positive here. The PCR product was sequenced and corresponded with the expected sequence for the plasmid. 3 of the 8 colonies were selected to be grown up and Plasmid Prepped to remove the plasmid and allow restriction digest and sequencing of the plasmid for further verification.<br><br />
<br />
<br><b>Restriction Digest</b><br><br />
<br />
The pUS203 plasmid was further confirmed with restriction digestion. Spe1, Pst1 and EcoR1 are in the prefix and suffix and should cut. An Nhe1 restriction site was added into the gBlock code and so a cut with this enzyme would indicate the gBlock had been inserted. Sac1 should not cut and should be identical to the uncut control lane. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/4/4e/USyd-AustraliaWell_Images_restriction_Digest.png"><br><br />
<br />
The digest results were as expected using SnapGene’s Virtual Digest tool. We were satisfied with these results despite the Sac1 error because the other fragments were as expected and most importantly because Nhe1 cut, which could only have occurred if the gblock had been incorporated into the plasmid. The Sac1 sample was heat-treated to inactive the Sac1 enzyme and used was used as a template to PCR the BioBrick. The results were still as expected which showed that the unexpected action of the Sac1 seen in the gel for the digest of this plasmid luckily didn’t reflect an error in the biobrick code. <br><br />
<br />
<b>Biobrick PCR and sequencing</b><br><br><br />
The final check for this plasmid was to PCR the entire biobrick fragment using iGEM16 and iGEM25 and sequencing the product. <br />
</p><br />
<br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<h2>Validation</h2><br />
<ul style="font-weight: normal;"><br />
To validate our araC-pBAD/IntI1 BioBrick it would need to be present in a low copy plasmid in order to reduce the amount produced as it is potentially toxic to the cell at high concentrations. Our intial plan was to insert the BioBrick into pUS202 but it turned out not to be low-copy after validation. The back up option was to ligate the BioBrick into pSB6A1. The initial attempt was not successful and due to time constraints we decided to try and test the integron construct in the pSB1C3 it was currently in even with the potential toxicity.<br><br><br />
To test for the proper expression of our integron gene in the BioBrick we wanted to have an AttI site and integrase protein present in E. coli. This was so that upon transformation with a cassette (which has an AttC site) a co-integrate would quickly form with the AttI site and before the cassettes could be lost out of the cell. Upon integration of the cassette the Gentomycin resistance (GmR) present on the cassette is expressed allowing us to select for the correct cells on Gentomycin plates. The negative control had no arabinose induction and so the cassette should not integrate and the cells should not be Gm resistant.<br><br><br />
<img src="https://static.igem.org/mediawiki/2014/7/71/USyd-Australia_Integrase_diagrams_1.png" width="98%"><br><br><br />
The initial experiments did not yield positive results and due to time constraints we have not been able to troubleshoot the experiment. <br />
</ul><br />
</li><br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
</html></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/pUS203Team:USyd-Australia/pUS2032014-10-18T02:54:55Z<p>Andy.Bachler: </p>
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<h2><a href="http://parts.igem.org/Part:BBa_K1388000">pUS203 Controllable Integrase production</a></h2><br />
</html><br />
<br />
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<p style="font-size:12px"><br />
To see this part on the iGEM Parts catalogue please click <a href="http://parts.igem.org/Part:BBa_K1388000">here</a>. <br>Sequences for this part are also available in <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLRDJ3OVdUUlhEVjA/view?usp=sharing">SnapGene (.dna file)</a>, <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLT3hQZno4VXFVZ0U/view?usp=sharing">FASTA (.fa file)</a>, <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLbndkclF5UlFNVE0/view?usp=sharing">GenBank Standard (.gb file)</a> and <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLak5Qdi1CbFZ4bm8/view?usp=sharing">Plain Text (.txt file)</a> file format.<br />
</p><br />
<h2>Aim</h2><br />
<p>Our aim was to create a controllable Integrase producing plasmid as a submission part. From the submission plasmid the controllable Integrase Part could then be ligated into a convenient low-copy plasmid to help reduce the potential toxicity of Integrase to the cell.</p><br />
<br />
<h2>Approach</h2><br />
<p><br />
<img src="https://static.igem.org/mediawiki/2014/6/67/PUS203.png" align="right" width="50%"><br />
The 2010 Paris Bettencourt team had developed a <a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K329013>controllable Integrase part</a> for use a population counter in their project. We contacted them to see if we could source the part from them but it was unfortunately not possible. We set about creating the BioBrick using the araC-pBAD from the well characterized part <a href=http://parts.igem.org/Part:BBa_K731201>K731201</a> and ordering the IntI1 as a gBlock. The araC-pBAD system in pSB1C3 had previously been prepared by Honours student Sam Ross (called the SamR construct). Our next steps were the design of the gBlock and integration of the IntI1 gBlock into the SamR construct.<br />
</p><br />
<br />
<h2>Materials</h2><br />
<ul>DNA<ul><br />
<li>SamR Construct: araC-pBAD in pSC1B3</li><br />
<li><a href="#">IntI1 gBlock</a>: to be inserted into SamR</li><br />
<li>pUS44: Verified plasmid containing an AttI site, used in Validation</li><br />
<li>aeBlue: Cassette PCR'ed from <a href=http://parts.igem.org/Part:BBa_K1033929>K1033929</a> for us in validation</li><br />
</ul><br />
</ul><br />
<br />
<ul>Primers<br />
<table><br />
<tr><br />
<td width="50%"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1413">iGEM1413</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1414">iGEM1414</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1415">iGEM1415</a></li><br />
</ul><br />
</td><br />
<td><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1416">iGEM1416</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers">iGEM16</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers">iGEM25</a></li><br />
</ul><br />
</td><br />
</tr><br />
<tr><br />
</table><br />
<br />
<h2>Inti1 gBlock Design</h2><br />
<p><br />
The code for the sequence for the IntI1 gene was sourced from pUS2056 but is highly conserved so this choice was out of convenience as the sequence was locally available. It was modified by Tom Geddes, Rokiah Alford and our supervisor Dr Nicholas Coleman to:<br><ul><br />
<li>Include a Sac1 restriction enzyme site at the beginning before the Inti1 gene so it could be excised independently of the promoter if needed</li><br />
<li>Include a different ribosome binding site </li><br />
<li>Include Gibson ends that overlapped with the backbone for simple Gibson Assembly later with the SamR construct.</li><br />
<li>Remove an illegal Spe1 site near the end to make it compliant with iGEM requirements</li><br />
<li>Remove a promoter pC that was coded in the opposite direction to the promoter we wanted to use and would have clashed with the controllable promotion of this part</li></ul><br><br />
Our first attempt to Gibson Assemble the SamR construct and IntI1 gBlock failed because there was extra code after the code for pBAD which we had not anticipated. This meant that the Gibson Assembly could not properly integrate the gBlock. We discussed numerous options to try and either get rid of the problem sequence on the SamR construct or modify the ends of the gBlock to accommodate the problem sequence. In the end it was decided that a re-design of the gBlock would be the most efficient way to solve the problem. <br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<h2>Inti1 gBlock ReDesign</h2><br />
<p><br />
Three major changes were decided: <br><br />
<ul><br />
<li>Changed the code for the ribosome cut site as a base pair was incorrect</li><br />
<li>To put an Nhe1 restriction enzyme cut site between the IntI1 gene and the promoters on the SamR construct. This was done to help in the ligation of this BioBrick into the low-copy backbone pUS202. </li><br />
<li>Changing of the end sequences to accommodate the extra code present on the SamR construct</li><br />
</ul><br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<br />
<h2>Construction</h2><br />
<p><br />
The IntI1 gBlock was inserted into the SamR construct using <a href="http://www.addgene.org/plasmid-protocols/gibson-assembly/">Gibson Asssembly</a>. The ends of the gBlock in the redesign were compatible with the ends of the SamR construct and after running the Gibson Assembly reaction we used a variety of methods to prove that our gBlock had inserted correctly.<br />
</p><br />
<br />
<p><br />
These changed were performed by our supervisor Nick Coleman in consultation with Rokiah Alford. The redesigned gBlock was able to be successfully Gibsoned into the SamR backbone. To verify the presence of the insert we used junction primers which would amplify over both ends of the insert and could only amplify if the gBlock had been integrated into the SamR construct. <br><br />
<br><b>Junction primer PCR</b><br><br />
<br />
Junction primers for the junction of promoter and gblock were iGEM1413 and iGEM1414 and were expected to give a fragment size of 377 base pairs. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/4/4e/USyd-AustraliaWell_Images_restriction_Digest.png"> <br><br />
<br />
9 out of the 12 colonies screened gave a positive result and these were further screened with a second set of junction primers on the other side of the gBlock. The junction primers for this were iGEM1416 and iGEM25 and were expected to yielad a fragment size of 432bp. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/7/70/USyd-Australia_RealSecondPCR_Junction_screen.png"> <br><br />
<br />
Again 9 of the 12 colonies were positive here. The PCR product was sequenced and corresponded with the expected sequence for the plasmid. 3 of the 8 colonies were selected to be grown up and Plasmid Prepped to remove the plasmid and allow restriction digest and sequencing of the plasmid for further verification.<br><br />
<br />
<br><b>Restriction Digest</b><br><br />
<br />
The pUS203 plasmid was further confirmed with restriction digestion. Spe1, Pst1 and EcoR1 are in the prefix and suffix and should cut. An Nhe1 restriction site was added into the gBlock code and so a cut with this enzyme would indicate the gBlock had been inserted. Sac1 should not cut and should be identical to the uncut control lane. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/4/4e/USyd-AustraliaWell_Images_restriction_Digest.png"><br><br />
<br />
The digest results were as expected using SnapGene’s Virtual Digest tool. We were satisfied with these results despite the Sac1 error because the other fragments were as expected and most importantly because Nhe1 cut, which could only have occurred if the gblock had been incorporated into the plasmid. The Sac1 sample was heat-treated to inactive the Sac1 enzyme and used was used as a template to PCR the BioBrick. The results were still as expected which showed that the unexpected action of the Sac1 seen in the gel for the digest of this plasmid luckily didn’t reflect an error in the biobrick code. <br><br />
<br />
<b>Biobrick PCR and sequencing</b><br><br />
The final check for this plasmid was to PCR the entire biobrick fragment using iGEM16 and iGEM25 and sequencing the product. <br />
</p><br />
<br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<h2>Validation</h2><br />
<ul style="font-weight: normal;"><br />
To validate our araC-pBAD/IntI1 BioBrick it would need to be present in a low copy plasmid in order to reduce the amount produced as it is potentially toxic to the cell at high concentrations. Our intial plan was to insert the BioBrick into pUS202 but it turned out not to be low-copy after validation. The back up option was to ligate the BioBrick into pSB6A1. The initial attempt was not successful and due to time constraints we decided to try and test the integron construct in the pSB1C3 it was currently in even with the potential toxicity.<br><br><br />
To test for the proper expression of our integron gene in the BioBrick we wanted to have an AttI site and integrase protein present in E. coli. This was so that upon transformation with a cassette (which has an AttC site) a co-integrate would quickly form with the AttI site and before the cassettes could be lost out of the cell. Upon integration of the cassette the Gentomycin resistance (GmR) present on the cassette is expressed allowing us to select for the correct cells on Gentomycin plates. The negative control had no arabinose induction and so the cassette should not integrate and the cells should not be Gm resistant.<br><br><br />
<img src="https://static.igem.org/mediawiki/2014/7/71/USyd-Australia_Integrase_diagrams_1.png" width="98%"><br><br><br />
The initial experiments did not yield positive results and due to time constraints we have not been able to troubleshoot the experiment. <br />
</ul><br />
</li><br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
</html></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/pUS203Team:USyd-Australia/pUS2032014-10-18T02:53:34Z<p>Andy.Bachler: </p>
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<h2><a href="http://parts.igem.org/Part:BBa_K1388000">pUS203 Controllable Integrase production</a></h2><br />
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<p style="font-size:12px"><br />
To see this part on the iGEM Parts catalogue please click <a href="http://parts.igem.org/Part:BBa_K1388000">here</a>. <br>Sequences for this part are also available in <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLRDJ3OVdUUlhEVjA/view?usp=sharing">SnapGene (.dna file)</a>, <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLT3hQZno4VXFVZ0U/view?usp=sharing">FASTA (.fa file)</a>, <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLbndkclF5UlFNVE0/view?usp=sharing">GenBank Standard (.gb file)</a> and <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLak5Qdi1CbFZ4bm8/view?usp=sharing">Plain Text (.txt file)</a> file format.<br />
</p><br />
<h2>Aim</h2><br />
<p>Our aim was to create a controllable Integrase producing plasmid as a submission part. From the submission plasmid the controllable Integrase Part could then be ligated into a convenient low-copy plasmid to help reduce the potential toxicity of Integrase to the cell.</p><br />
<br />
<h2>Approach</h2><br />
<p><br />
<img src="https://static.igem.org/mediawiki/2014/6/67/PUS203.png" align="right" width="50%"><br />
The 2010 Paris Bettencourt team had developed a <a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K329013>controllable Integrase part</a> for use a population counter in their project. We contacted them to see if we could source the part from them but it was unfortunately not possible. We set about creating the BioBrick using the araC-pBAD from the well characterized part <a href=http://parts.igem.org/Part:BBa_K731201>K731201</a> and ordering the IntI1 as a gBlock. The araC-pBAD system in pSB1C3 had previously been prepared by Honours student Sam Ross (called the SamR construct). Our next steps were the design of the gBlock and integration of the IntI1 gBlock into the SamR construct.<br />
</p><br />
<br />
<h2>Materials</h2><br />
<ul>DNA<ul><br />
<li>SamR Construct: araC-pBAD in pSC1B3</li><br />
<li><a href="#">IntI1 gBlock</a>: to be inserted into SamR</li><br />
<li>pUS44: Verified plasmid containing an AttI site, used in Validation</li><br />
<li>aeBlue: Cassette PCR'ed from <a href=http://parts.igem.org/Part:BBa_K1033929>K1033929</a> for us in validation</li><br />
</ul><br />
</ul><br />
<br />
<ul>Primers<br />
<table><br />
<tr><br />
<td width="50%"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1413">iGEM1413</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1414">iGEM1414</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1415">iGEM1415</a></li><br />
</ul><br />
</td><br />
<td><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1416">iGEM1416</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers">iGEM16</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers">iGEM25</a></li><br />
</ul><br />
</td><br />
</tr><br />
<tr><br />
</table><br />
<br />
<h2>Inti1 gBlock Design</h2><br />
<p><br />
The code for the sequence for the IntI1 gene was sourced from pUS2056 but is highly conserved so this choice was out of convenience as the sequence was locally available. It was modified by Tom Geddes, Rokiah Alford and our supervisor Dr Nicholas Coleman to:<br><ul><br />
<li>Include a Sac1 restriction enzyme site at the beginning before the Inti1 gene so it could be excised independently of the promoter if needed</li><br />
<li>Include a different ribosome binding site </li><br />
<li>Include Gibson ends that overlapped with the backbone for simple Gibson Assembly later with the SamR construct.</li><br />
<li>Remove an illegal Spe1 site near the end to make it compliant with iGEM requirements</li><br />
<li>Remove a promoter pC that was coded in the opposite direction to the promoter we wanted to use and would have clashed with the controllable promotion of this part</li></ul><br><br />
Our first attempt to Gibson Assemble the SamR construct and IntI1 gBlock failed because there was extra code after the code for pBAD which we had not anticipated. This meant that the Gibson Assembly could not properly integrate the gBlock. We discussed numerous options to try and either get rid of the problem sequence on the SamR construct or modify the ends of the gBlock to accommodate the problem sequence. In the end it was decided that a re-design of the gBlock would be the most efficient way to solve the problem. <br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<h2>Inti1 gBlock ReDesign</h2><br />
<p><br />
Three major changes were decided: <br><br />
<ul><br />
<li>Changed the code for the ribosome cut site as a base pair was incorrect</li><br />
<li>To put an Nhe1 restriction enzyme cut site between the IntI1 gene and the promoters on the SamR construct. This was done to help in the ligation of this BioBrick into the low-copy backbone pUS202. </li><br />
<li>Changing of the end sequences to accommodate the extra code present on the SamR construct</li><br />
</ul><br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<br />
<h2>Construction</h2><br />
<p><br />
The IntI1 gBlock was inserted into the SamR construct using <a href="http://www.addgene.org/plasmid-protocols/gibson-assembly/">Gibson Asssembly</a>. The ends of the gBlock in the redesign were compatible with the ends of the SamR construct and after running the Gibson Assembly reaction we used a variety of methods to prove that our gBlock had inserted correctly.<br />
</p><br />
<br />
<p><br />
These changed were performed by our supervisor Nick Coleman in consultation with Rokiah Alford. The redesigned gBlock was able to be successfully Gibsoned into the SamR backbone. To verify the presence of the insert we used junction primers which would amplify over both ends of the insert and could only amplify if the gBlock had been integrated into the SamR construct. <br><br />
<br><b>Junction primer PCR</b><br><br />
<br />
Junction primers for the junction of promoter and gblock were iGEM1413 and iGEM1414 and were expected to give a fragment size of 377 base pairs. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/4/4e/USyd-AustraliaWell_Images_restriction_Digest.png"> <br><br />
<br />
9 out of the 12 colonies screened gave a positive result and these were further screened with a second set of junction primers on the other side of the gBlock. The junction primers for this were iGEM1416 and iGEM25 and were expected to yielad a fragment size of 432bp. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/7/70/USyd-Australia_RealSecondPCR_Junction_screen.png"> <br><br />
<br />
Again 9 of the 12 colonies were positive here. The PCR product was sequenced and corresponded with the expected sequence for the plasmid. 3 of the 8 colonies were selected to be grown up and Plasmid Prepped to remove the plasmid and allow restriction digest and sequencing of the plasmid for further verification.<br><br />
<br />
<br><b>Restriction Digest</b><br><br />
<br />
The pUS203 plasmid was further confirmed with restriction digestion. Spe1, Pst1 and EcoR1 are in the prefix and suffix and should cut. An Nhe1 restriction site was added into the gBlock code and so a cut with this enzyme would indicate the gBlock had been inserted. Sac1 should not cut and should be identical to the uncut control lane. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/4/4e/USyd-AustraliaWell_Images_restriction_Digest.png"><br><br />
<br />
The digest results were as expected using SnapGene’s Virtual Digest tool. We were satisfied with these results despite the Sac1 error because the other fragments were as expected and most importantly because Nhe1 cut, which could only have occurred if the gblock had been incorporated into the plasmid. The Sac1 sample was heat-treated to inactive the Sac1 enzyme and used was used as a template to PCR the BioBrick. The results were still as expected which showed that the unexpected action of the Sac1 seen in the gel for the digest of this plasmid luckily didn’t reflect an error in the biobrick code. <br />
<b>Biobrick PCR and sequencing</b><br />
The final check for this plasmid was to PCR the entire biobrick fragment using iGEM16 and iGEM25 and sequencing the product. <br />
</p><br />
<br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<h2>Validation</h2><br />
<ul style="font-weight: normal;"><br />
To validate our araC-pBAD/IntI1 BioBrick it would need to be present in a low copy plasmid in order to reduce the amount produced as it is potentially toxic to the cell at high concentrations. Our intial plan was to insert the BioBrick into pUS202 but it turned out not to be low-copy after validation. The back up option was to ligate the BioBrick into pSB6A1. The initial attempt was not successful and due to time constraints we decided to try and test the integron construct in the pSB1C3 it was currently in even with the potential toxicity.<br><br><br />
To test for the proper expression of our integron gene in the BioBrick we wanted to have an AttI site and integrase protein present in E. coli. This was so that upon transformation with a cassette (which has an AttC site) a co-integrate would quickly form with the AttI site and before the cassettes could be lost out of the cell. Upon integration of the cassette the Gentomycin resistance (GmR) present on the cassette is expressed allowing us to select for the correct cells on Gentomycin plates. The negative control had no arabinose induction and so the cassette should not integrate and the cells should not be Gm resistant.<br><br><br />
<img src="https://static.igem.org/mediawiki/2014/7/71/USyd-Australia_Integrase_diagrams_1.png" width="98%"><br><br><br />
The initial experiments did not yield positive results and due to time constraints we have not been able to troubleshoot the experiment. <br />
</ul><br />
</li><br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
</html></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/pUS203Team:USyd-Australia/pUS2032014-10-18T02:52:53Z<p>Andy.Bachler: </p>
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<h2><a href="http://parts.igem.org/Part:BBa_K1388000">pUS203 Controllable Integrase production</a></h2><br />
</html><br />
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<p style="font-size:12px"><br />
To see this part on the iGEM Parts catalogue please click <a href="http://parts.igem.org/Part:BBa_K1388000">here</a>. <br>Sequences for this part are also available in <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLRDJ3OVdUUlhEVjA/view?usp=sharing">SnapGene (.dna file)</a>, <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLT3hQZno4VXFVZ0U/view?usp=sharing">FASTA (.fa file)</a>, <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLbndkclF5UlFNVE0/view?usp=sharing">GenBank Standard (.gb file)</a> and <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLak5Qdi1CbFZ4bm8/view?usp=sharing">Plain Text (.txt file)</a> file format.<br />
</p><br />
<h2>Aim</h2><br />
<p>Our aim was to create a controllable Integrase producing plasmid as a submission part. From the submission plasmid the controllable Integrase Part could then be ligated into a convenient low-copy plasmid to help reduce the potential toxicity of Integrase to the cell.</p><br />
<br />
<h2>Approach</h2><br />
<p><br />
<img src="https://static.igem.org/mediawiki/2014/6/67/PUS203.png" align="right" width="50%"><br />
The 2010 Paris Bettencourt team had developed a <a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K329013>controllable Integrase part</a> for use a population counter in their project. We contacted them to see if we could source the part from them but it was unfortunately not possible. We set about creating the BioBrick using the araC-pBAD from the well characterized part <a href=http://parts.igem.org/Part:BBa_K731201>K731201</a> and ordering the IntI1 as a gBlock. The araC-pBAD system in pSB1C3 had previously been prepared by Honours student Sam Ross (called the SamR construct). Our next steps were the design of the gBlock and integration of the IntI1 gBlock into the SamR construct.<br />
</p><br />
<br />
<h2>Materials</h2><br />
<ul>DNA<ul><br />
<li>SamR Construct: araC-pBAD in pSC1B3</li><br />
<li><a href="#">IntI1 gBlock</a>: to be inserted into SamR</li><br />
<li>pUS44: Verified plasmid containing an AttI site, used in Validation</li><br />
<li>aeBlue: Cassette PCR'ed from <a href=http://parts.igem.org/Part:BBa_K1033929>K1033929</a> for us in validation</li><br />
</ul><br />
</ul><br />
<br />
<ul>Primers<br />
<table><br />
<tr><br />
<td width="50%"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1413">iGEM1413</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1414">iGEM1414</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1416">iGEM1416</a></li></ul><br />
</td><br />
<td><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers">iGEM16</a></li><br />
<li><a href="https://2014.igem.org/Team:USyd-Australia/Notebook/Primers">iGEM25</a></li><br />
</ul><br />
</td><br />
</tr><br />
<tr><br />
</table><br />
<br />
<h2>Inti1 gBlock Design</h2><br />
<p><br />
The code for the sequence for the IntI1 gene was sourced from pUS2056 but is highly conserved so this choice was out of convenience as the sequence was locally available. It was modified by Tom Geddes, Rokiah Alford and our supervisor Dr Nicholas Coleman to:<br><ul><br />
<li>Include a Sac1 restriction enzyme site at the beginning before the Inti1 gene so it could be excised independently of the promoter if needed</li><br />
<li>Include a different ribosome binding site </li><br />
<li>Include Gibson ends that overlapped with the backbone for simple Gibson Assembly later with the SamR construct.</li><br />
<li>Remove an illegal Spe1 site near the end to make it compliant with iGEM requirements</li><br />
<li>Remove a promoter pC that was coded in the opposite direction to the promoter we wanted to use and would have clashed with the controllable promotion of this part</li></ul><br><br />
Our first attempt to Gibson Assemble the SamR construct and IntI1 gBlock failed because there was extra code after the code for pBAD which we had not anticipated. This meant that the Gibson Assembly could not properly integrate the gBlock. We discussed numerous options to try and either get rid of the problem sequence on the SamR construct or modify the ends of the gBlock to accommodate the problem sequence. In the end it was decided that a re-design of the gBlock would be the most efficient way to solve the problem. <br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<h2>Inti1 gBlock ReDesign</h2><br />
<p><br />
Three major changes were decided: <br><br />
<ul><br />
<li>Changed the code for the ribosome cut site as a base pair was incorrect</li><br />
<li>To put an Nhe1 restriction enzyme cut site between the IntI1 gene and the promoters on the SamR construct. This was done to help in the ligation of this BioBrick into the low-copy backbone pUS202. </li><br />
<li>Changing of the end sequences to accommodate the extra code present on the SamR construct</li><br />
</ul><br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<br />
<h2>Construction</h2><br />
<p><br />
The IntI1 gBlock was inserted into the SamR construct using <a href="http://www.addgene.org/plasmid-protocols/gibson-assembly/">Gibson Asssembly</a>. The ends of the gBlock in the redesign were compatible with the ends of the SamR construct and after running the Gibson Assembly reaction we used a variety of methods to prove that our gBlock had inserted correctly.<br />
</p><br />
<br />
<p><br />
These changed were performed by our supervisor Nick Coleman in consultation with Rokiah Alford. The redesigned gBlock was able to be successfully Gibsoned into the SamR backbone. To verify the presence of the insert we used junction primers which would amplify over both ends of the insert and could only amplify if the gBlock had been integrated into the SamR construct. <br><br />
<br><b>Junction primer PCR</b><br><br />
<br />
Junction primers for the junction of promoter and gblock were iGEM1413 and iGEM1414 and were expected to give a fragment size of 377 base pairs. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/4/4e/USyd-AustraliaWell_Images_restriction_Digest.png"> <br><br />
<br />
9 out of the 12 colonies screened gave a positive result and these were further screened with a second set of junction primers on the other side of the gBlock. The junction primers for this were iGEM1416 and iGEM25 and were expected to yielad a fragment size of 432bp. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/7/70/USyd-Australia_RealSecondPCR_Junction_screen.png"> <br><br />
<br />
Again 9 of the 12 colonies were positive here. The PCR product was sequenced and corresponded with the expected sequence for the plasmid. 3 of the 8 colonies were selected to be grown up and Plasmid Prepped to remove the plasmid and allow restriction digest and sequencing of the plasmid for further verification.<br><br />
<br />
<br><b>Restriction Digest</b><br><br />
<br />
The pUS203 plasmid was further confirmed with restriction digestion. Spe1, Pst1 and EcoR1 are in the prefix and suffix and should cut. An Nhe1 restriction site was added into the gBlock code and so a cut with this enzyme would indicate the gBlock had been inserted. Sac1 should not cut and should be identical to the uncut control lane. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/4/4e/USyd-AustraliaWell_Images_restriction_Digest.png"><br><br />
<br />
The digest results were as expected using SnapGene’s Virtual Digest tool. We were satisfied with these results despite the Sac1 error because the other fragments were as expected and most importantly because Nhe1 cut, which could only have occurred if the gblock had been incorporated into the plasmid. The Sac1 sample was heat-treated to inactive the Sac1 enzyme and used was used as a template to PCR the BioBrick. The results were still as expected which showed that the unexpected action of the Sac1 seen in the gel for the digest of this plasmid luckily didn’t reflect an error in the biobrick code. <br />
<b>Biobrick PCR and sequencing</b><br />
The final check for this plasmid was to PCR the entire biobrick fragment using iGEM16 and iGEM25 and sequencing the product. <br />
</p><br />
<br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<h2>Validation</h2><br />
<ul style="font-weight: normal;"><br />
To validate our araC-pBAD/IntI1 BioBrick it would need to be present in a low copy plasmid in order to reduce the amount produced as it is potentially toxic to the cell at high concentrations. Our intial plan was to insert the BioBrick into pUS202 but it turned out not to be low-copy after validation. The back up option was to ligate the BioBrick into pSB6A1. The initial attempt was not successful and due to time constraints we decided to try and test the integron construct in the pSB1C3 it was currently in even with the potential toxicity.<br><br><br />
To test for the proper expression of our integron gene in the BioBrick we wanted to have an AttI site and integrase protein present in E. coli. This was so that upon transformation with a cassette (which has an AttC site) a co-integrate would quickly form with the AttI site and before the cassettes could be lost out of the cell. Upon integration of the cassette the Gentomycin resistance (GmR) present on the cassette is expressed allowing us to select for the correct cells on Gentomycin plates. The negative control had no arabinose induction and so the cassette should not integrate and the cells should not be Gm resistant.<br><br><br />
<img src="https://static.igem.org/mediawiki/2014/7/71/USyd-Australia_Integrase_diagrams_1.png" width="98%"><br><br><br />
The initial experiments did not yield positive results and due to time constraints we have not been able to troubleshoot the experiment. <br />
</ul><br />
</li><br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
</html></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/pUS203Team:USyd-Australia/pUS2032014-10-18T02:51:36Z<p>Andy.Bachler: </p>
<hr />
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<br />
<html><br />
<a name="protstart"></a><br />
<h2><a href="http://parts.igem.org/Part:BBa_K1388000">pUS203 Controllable Integrase production</a></h2><br />
</html><br />
<br />
<html><br />
<br />
<script type="text/javascript"><br />
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<br />
<p style="font-size:12px"><br />
To see this part on the iGEM Parts catalogue please click <a href="http://parts.igem.org/Part:BBa_K1388000">here</a>. <br>Sequences for this part are also available in <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLRDJ3OVdUUlhEVjA/view?usp=sharing">SnapGene (.dna file)</a>, <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLT3hQZno4VXFVZ0U/view?usp=sharing">FASTA (.fa file)</a>, <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLbndkclF5UlFNVE0/view?usp=sharing">GenBank Standard (.gb file)</a> and <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLak5Qdi1CbFZ4bm8/view?usp=sharing">Plain Text (.txt file)</a> file format.<br />
</p><br />
<h2>Aim</h2><br />
<p>Our aim was to create a controllable Integrase producing plasmid as a submission part. From the submission plasmid the controllable Integrase Part could then be ligated into a convenient low-copy plasmid to help reduce the potential toxicity of Integrase to the cell.</p><br />
<br />
<h2>Approach</h2><br />
<p><br />
<img src="https://static.igem.org/mediawiki/2014/6/67/PUS203.png" align="right" width="50%"><br />
The 2010 Paris Bettencourt team had developed a <a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K329013>controllable Integrase part</a> for use a population counter in their project. We contacted them to see if we could source the part from them but it was unfortunately not possible. We set about creating the BioBrick using the araC-pBAD from the well characterized part <a href=http://parts.igem.org/Part:BBa_K731201>K731201</a> and ordering the IntI1 as a gBlock. The araC-pBAD system in pSB1C3 had previously been prepared by Honours student Sam Ross (called the SamR construct). Our next steps were the design of the gBlock and integration of the IntI1 gBlock into the SamR construct.<br />
</p><br />
<br />
<h2>Materials</h2><br />
<ul>DNA<ul><br />
<li>SamR Construct: araC-pBAD in pSC1B3</li><br />
<li><a href="#">IntI1 gBlock</a>: to be inserted into SamR</li><br />
<li>pUS44: Verified plasmid containing an AttI site, used in Validation</li><br />
<li>aeBlue: Cassette PCR'ed from <a href=http://parts.igem.org/Part:BBa_K1033929>K1033929</a> for us in validation</li><br />
</ul><br />
</ul><br />
<br />
<ul>Primers<br />
<table><br />
<tr><br />
<td width="50%"><br />
<ul><br />
<li><a href=”https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1413”>iGEM1413</a></li><br />
<li><a href=”https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1414”>iGEM1414</a></li><br />
<li><a href=”https://2014.igem.org/Team:USyd-Australia/Notebook/Primers#iGEM1416”>iGEM1416</a></li></ul><br />
</td><br />
<td><br />
<ul><br />
<li><a href=”https://2014.igem.org/Team:USyd-Australia/Notebook/Primers”>iGEM16</a></li><br />
<li><a href=”https://2014.igem.org/Team:USyd-Australia/Notebook/Primers”>iGEM25</a></li><br />
</ul><br />
</td><br />
</tr><br />
<tr><br />
</table><br />
<br />
<h2>Inti1 gBlock Design</h2><br />
<p><br />
The code for the sequence for the IntI1 gene was sourced from pUS2056 but is highly conserved so this choice was out of convenience as the sequence was locally available. It was modified by Tom Geddes, Rokiah Alford and our supervisor Dr Nicholas Coleman to:<br><ul><br />
<li>Include a Sac1 restriction enzyme site at the beginning before the Inti1 gene so it could be excised independently of the promoter if needed</li><br />
<li>Include a different ribosome binding site </li><br />
<li>Include Gibson ends that overlapped with the backbone for simple Gibson Assembly later with the SamR construct.</li><br />
<li>Remove an illegal Spe1 site near the end to make it compliant with iGEM requirements</li><br />
<li>Remove a promoter pC that was coded in the opposite direction to the promoter we wanted to use and would have clashed with the controllable promotion of this part</li></ul><br><br />
Our first attempt to Gibson Assemble the SamR construct and IntI1 gBlock failed because there was extra code after the code for pBAD which we had not anticipated. This meant that the Gibson Assembly could not properly integrate the gBlock. We discussed numerous options to try and either get rid of the problem sequence on the SamR construct or modify the ends of the gBlock to accommodate the problem sequence. In the end it was decided that a re-design of the gBlock would be the most efficient way to solve the problem. <br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<h2>Inti1 gBlock ReDesign</h2><br />
<p><br />
Three major changes were decided: <br><br />
<ul><br />
<li>Changed the code for the ribosome cut site as a base pair was incorrect</li><br />
<li>To put an Nhe1 restriction enzyme cut site between the IntI1 gene and the promoters on the SamR construct. This was done to help in the ligation of this BioBrick into the low-copy backbone pUS202. </li><br />
<li>Changing of the end sequences to accommodate the extra code present on the SamR construct</li><br />
</ul><br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<br />
<h2>Construction</h2><br />
<p><br />
The IntI1 gBlock was inserted into the SamR construct using <a href="http://www.addgene.org/plasmid-protocols/gibson-assembly/">Gibson Asssembly</a>. The ends of the gBlock in the redesign were compatible with the ends of the SamR construct and after running the Gibson Assembly reaction we used a variety of methods to prove that our gBlock had inserted correctly.<br />
</p><br />
<br />
<p><br />
These changed were performed by our supervisor Nick Coleman in consultation with Rokiah Alford. The redesigned gBlock was able to be successfully Gibsoned into the SamR backbone. To verify the presence of the insert we used junction primers which would amplify over both ends of the insert and could only amplify if the gBlock had been integrated into the SamR construct. <br><br />
<br><b>Junction primer PCR</b><br><br />
<br />
Junction primers for the junction of promoter and gblock were iGEM1413 and iGEM1414 and were expected to give a fragment size of 377 base pairs. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/4/4e/USyd-AustraliaWell_Images_restriction_Digest.png"> <br><br />
<br />
9 out of the 12 colonies screened gave a positive result and these were further screened with a second set of junction primers on the other side of the gBlock. The junction primers for this were iGEM1416 and iGEM25 and were expected to yielad a fragment size of 432bp. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/7/70/USyd-Australia_RealSecondPCR_Junction_screen.png"> <br><br />
<br />
Again 9 of the 12 colonies were positive here. The PCR product was sequenced and corresponded with the expected sequence for the plasmid. 3 of the 8 colonies were selected to be grown up and Plasmid Prepped to remove the plasmid and allow restriction digest and sequencing of the plasmid for further verification.<br><br />
<br />
<br><b>Restriction Digest</b><br><br />
<br />
The pUS203 plasmid was further confirmed with restriction digestion. Spe1, Pst1 and EcoR1 are in the prefix and suffix and should cut. An Nhe1 restriction site was added into the gBlock code and so a cut with this enzyme would indicate the gBlock had been inserted. Sac1 should not cut and should be identical to the uncut control lane. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/4/4e/USyd-AustraliaWell_Images_restriction_Digest.png"><br><br />
<br />
The digest results were as expected using SnapGene’s Virtual Digest tool. We were satisfied with these results despite the Sac1 error because the other fragments were as expected and most importantly because Nhe1 cut, which could only have occurred if the gblock had been incorporated into the plasmid. The Sac1 sample was heat-treated to inactive the Sac1 enzyme and used was used as a template to PCR the BioBrick. The results were still as expected which showed that the unexpected action of the Sac1 seen in the gel for the digest of this plasmid luckily didn’t reflect an error in the biobrick code. <br />
<b>Biobrick PCR and sequencing</b><br />
The final check for this plasmid was to PCR the entire biobrick fragment using iGEM16 and iGEM25 and sequencing the product. <br />
</p><br />
<br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<h2>Validation</h2><br />
<ul style="font-weight: normal;"><br />
To validate our araC-pBAD/IntI1 BioBrick it would need to be present in a low copy plasmid in order to reduce the amount produced as it is potentially toxic to the cell at high concentrations. Our intial plan was to insert the BioBrick into pUS202 but it turned out not to be low-copy after validation. The back up option was to ligate the BioBrick into pSB6A1. The initial attempt was not successful and due to time constraints we decided to try and test the integron construct in the pSB1C3 it was currently in even with the potential toxicity.<br><br><br />
To test for the proper expression of our integron gene in the BioBrick we wanted to have an AttI site and integrase protein present in E. coli. This was so that upon transformation with a cassette (which has an AttC site) a co-integrate would quickly form with the AttI site and before the cassettes could be lost out of the cell. Upon integration of the cassette the Gentomycin resistance (GmR) present on the cassette is expressed allowing us to select for the correct cells on Gentomycin plates. The negative control had no arabinose induction and so the cassette should not integrate and the cells should not be Gm resistant.<br><br><br />
<img src="https://static.igem.org/mediawiki/2014/7/71/USyd-Australia_Integrase_diagrams_1.png" width="98%"><br><br><br />
The initial experiments did not yield positive results and due to time constraints we have not been able to troubleshoot the experiment. <br />
</ul><br />
</li><br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
</html></div>Andy.Bachlerhttp://2014.igem.org/Template:Team:USyd-Australia/Template:StyleTemplate:Team:USyd-Australia/Template:Style2014-10-18T02:47:38Z<p>Andy.Bachler: </p>
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</html></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:46:48Z<p>Andy.Bachler: </p>
<hr />
<div>{{Team:USyd-Australia/Template:Style}}<br />
{{:Team:USyd-Australia/Template:Banner}}<br />
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<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template.</p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table id="gBlockTable"><br />
<br />
<tr id="gBlockTableRow"><th width="10%">Name</th><th width="45%">Sequence (5'-3')</th><th width="5%">Size</th><th width="20%">Purpose</th></tr><br />
<br />
<tr id="gBlockTableRow"><td id="gBlockTableColumn"><a name="IntI1.2"></a>IntI1</td><td id="gBlockTableColumn" style="word-wrap: break-word; font-size: 8px;">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td id="gBlockTableColumn">1121 bp</td><td id="gBlockTableColumn">To be inserted into SamR construct as part of the controllable integrase expression system</td></tr><br />
<br />
<tr id="gBlockTableRow"><td id="gBlockTableColumn"><a name="AttC-aeBlue-aacC1 "></a>AttC-aeBlue-aacC1 </td><td id="gBlockTableColumn" style="word-wrap: break-word; font-size: 8px;">TTTGAATTCGCGGCCGCTTCTAGAGTACCGAGGGACAGACTACCAACTCACAGCTAGCGCCTAACAATTCGTCCAAGCCGACGCCGCTTCGCGGCGCGGCTTAACTCAGGTGTTAGGCAAAGGAGGTTATTGATATGGCTTCACTGGTTAAAAAAGACATGTGCATCAAAATGACGATGGAAGGAACAGTAAACGGTCACCATTTCAAGTGTGTAGGAGAAGGCGAAGGCAAACCATTTGAAGGGACCCAGGTGGAAAAGATACGCATCACTGAAGGTGGGCCCTTACCATTTGCGTATGATATTTTGGCCCCTTGTTGCATGTATGGCAGTAAAACCTTCATTAAGCATGTGTCGGGTATTCCGGATTACTTTAAGGAGTCTTTTCCTGAGGGCTTTACCTGGGAAAGAACACAAATCTTCGAGGATGGCGGCTATCTCACCATACACCAGGACACGAGCCTTCAGGGTAATAATTTTATTTTCAAAGTTAATGTCATCGGTGCCAACTTCCCTGCAAACGGTCCCGTGATGCAGAAAAAAACAGCTGGATGGGAACCGTGCGTTGAGATGCTTTATCCGCGGGACGGCGTCCTGTGTGGTCAGAGCCTGATGGCCCTGAAATGCACTGATGGCAATCATCTGACGTCCCACCTGCGCACTACCTATCGTTCTCGCAAGCCATCCAATGCAGTTAACATGCCGGAATTTCATTTTGGGGATCATCGCATTGAGATTTTGAAAGCTGAACAAGGTAAATTTTATGAACAATACGAGTCAGCGGTGGCCCGTTACTGTGAGGCGGCACCGAGTAAATTAGGGCATCACTAATAAAGGATCCAAAGGAGGCTCAAGTATGGGCATCATTCGCACATGTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGATCTATATCTATGATCTCGCAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCGCGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGGAAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAATAATACTAGTAGCGGCCGCTGCAGCCC</td><td id="gBlockTableColumn">1347 bp</td><td id="gBlockTableColumn">To be used as a template for the creation of gene cassettes with antibiotic resistance and phenotypic markers</td></tr><br />
<br />
<tr id="gBlockTableRow"><td id="gBlockTableColumn"><a name="LacI-lacO-pLac-attI"></a>LacI-lacO-pLac-attI </td><td id="gBlockTableColumn" style="word-wrap: break-word; font-size: 8px;">GGGAATTCAAATCTAGAGACACCATCGAATGGTGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAATGAAGGAGGTGAATATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTCAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTAATAAGCGCAAAAAAAAAACCCCGCCCCTGACAGGGCGGGGTTTTTTTTCAACGCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAGGCAAAACTAGTAAACTGCAGGG</td><td id="gBlockTableColumn">1403 bp</td><td id="gBlockTableColumn">To be used as an AttI site with a Lac operon to allow controllable expression</td></tr><br />
</table><br />
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{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:46:25Z<p>Andy.Bachler: </p>
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<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template.</p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table id="gBlockTable"><br />
<br />
<tr id="gBlockTableRow"><th width="10%">Name</th><th width="45%">Sequence (5'-3')</th><th width="5%">Size</th><th width="20%">Purpose</th></tr><br />
<br />
<tr><td id="gBlockTableColumn"><a name="IntI1.2"></a>IntI1</td><td id="gBlockTableColumn" style="word-wrap: break-word; font-size: 8px;">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td id="gBlockTableColumn">1121 bp</td><td id="gBlockTableColumn">To be inserted into SamR construct as part of the controllable integrase expression system</td></tr><br />
<br />
<tr><td id="gBlockTableColumn"><a name="AttC-aeBlue-aacC1 "></a>AttC-aeBlue-aacC1 </td><td id="gBlockTableColumn" style="word-wrap: break-word; font-size: 8px;">TTTGAATTCGCGGCCGCTTCTAGAGTACCGAGGGACAGACTACCAACTCACAGCTAGCGCCTAACAATTCGTCCAAGCCGACGCCGCTTCGCGGCGCGGCTTAACTCAGGTGTTAGGCAAAGGAGGTTATTGATATGGCTTCACTGGTTAAAAAAGACATGTGCATCAAAATGACGATGGAAGGAACAGTAAACGGTCACCATTTCAAGTGTGTAGGAGAAGGCGAAGGCAAACCATTTGAAGGGACCCAGGTGGAAAAGATACGCATCACTGAAGGTGGGCCCTTACCATTTGCGTATGATATTTTGGCCCCTTGTTGCATGTATGGCAGTAAAACCTTCATTAAGCATGTGTCGGGTATTCCGGATTACTTTAAGGAGTCTTTTCCTGAGGGCTTTACCTGGGAAAGAACACAAATCTTCGAGGATGGCGGCTATCTCACCATACACCAGGACACGAGCCTTCAGGGTAATAATTTTATTTTCAAAGTTAATGTCATCGGTGCCAACTTCCCTGCAAACGGTCCCGTGATGCAGAAAAAAACAGCTGGATGGGAACCGTGCGTTGAGATGCTTTATCCGCGGGACGGCGTCCTGTGTGGTCAGAGCCTGATGGCCCTGAAATGCACTGATGGCAATCATCTGACGTCCCACCTGCGCACTACCTATCGTTCTCGCAAGCCATCCAATGCAGTTAACATGCCGGAATTTCATTTTGGGGATCATCGCATTGAGATTTTGAAAGCTGAACAAGGTAAATTTTATGAACAATACGAGTCAGCGGTGGCCCGTTACTGTGAGGCGGCACCGAGTAAATTAGGGCATCACTAATAAAGGATCCAAAGGAGGCTCAAGTATGGGCATCATTCGCACATGTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGATCTATATCTATGATCTCGCAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCGCGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGGAAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAATAATACTAGTAGCGGCCGCTGCAGCCC</td><td id="gBlockTableColumn">1347 bp</td><td id="gBlockTableColumn">To be used as a template for the creation of gene cassettes with antibiotic resistance and phenotypic markers</td></tr><br />
<br />
<tr><td id="gBlockTableColumn"><a name="LacI-lacO-pLac-attI"></a>LacI-lacO-pLac-attI </td><td id="gBlockTableColumn" style="word-wrap: break-word; font-size: 8px;">GGGAATTCAAATCTAGAGACACCATCGAATGGTGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAATGAAGGAGGTGAATATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTCAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTAATAAGCGCAAAAAAAAAACCCCGCCCCTGACAGGGCGGGGTTTTTTTTCAACGCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAGGCAAAACTAGTAAACTGCAGGG</td><td id="gBlockTableColumn">1403 bp</td><td id="gBlockTableColumn">To be used as an AttI site with a Lac operon to allow controllable expression</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Template:Team:USyd-Australia/Template:StyleTemplate:Team:USyd-Australia/Template:Style2014-10-18T02:42:53Z<p>Andy.Bachler: </p>
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</html></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:41:20Z<p>Andy.Bachler: </p>
<hr />
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{{:Team:USyd-Australia/Template:Banner}}<br />
<br />
<html><br />
<br />
<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template.</p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table id="gBlockTable"><br />
<br />
<tr><th width="10%">Name</th><th width="45%">Sequence (5'-3')</th><th width="5%">Size</th><th width="20%">Purpose</th></tr><br />
<br />
<tr><td><a name="IntI1.2"></a>IntI1</td><td style="word-wrap: break-word; font-size: 8px;">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td>1121 bp</td><td>To be inserted into SamR construct as part of the controllable integrase expression system</td></tr><br />
<br />
<tr><td><a name="AttC-aeBlue-aacC1 "></a>AttC-aeBlue-aacC1 </td><td style="word-wrap: break-word; font-size: 8px;">TTTGAATTCGCGGCCGCTTCTAGAGTACCGAGGGACAGACTACCAACTCACAGCTAGCGCCTAACAATTCGTCCAAGCCGACGCCGCTTCGCGGCGCGGCTTAACTCAGGTGTTAGGCAAAGGAGGTTATTGATATGGCTTCACTGGTTAAAAAAGACATGTGCATCAAAATGACGATGGAAGGAACAGTAAACGGTCACCATTTCAAGTGTGTAGGAGAAGGCGAAGGCAAACCATTTGAAGGGACCCAGGTGGAAAAGATACGCATCACTGAAGGTGGGCCCTTACCATTTGCGTATGATATTTTGGCCCCTTGTTGCATGTATGGCAGTAAAACCTTCATTAAGCATGTGTCGGGTATTCCGGATTACTTTAAGGAGTCTTTTCCTGAGGGCTTTACCTGGGAAAGAACACAAATCTTCGAGGATGGCGGCTATCTCACCATACACCAGGACACGAGCCTTCAGGGTAATAATTTTATTTTCAAAGTTAATGTCATCGGTGCCAACTTCCCTGCAAACGGTCCCGTGATGCAGAAAAAAACAGCTGGATGGGAACCGTGCGTTGAGATGCTTTATCCGCGGGACGGCGTCCTGTGTGGTCAGAGCCTGATGGCCCTGAAATGCACTGATGGCAATCATCTGACGTCCCACCTGCGCACTACCTATCGTTCTCGCAAGCCATCCAATGCAGTTAACATGCCGGAATTTCATTTTGGGGATCATCGCATTGAGATTTTGAAAGCTGAACAAGGTAAATTTTATGAACAATACGAGTCAGCGGTGGCCCGTTACTGTGAGGCGGCACCGAGTAAATTAGGGCATCACTAATAAAGGATCCAAAGGAGGCTCAAGTATGGGCATCATTCGCACATGTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGATCTATATCTATGATCTCGCAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCGCGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGGAAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAATAATACTAGTAGCGGCCGCTGCAGCCC</td><td>1347 bp</td><td>To be used as a template for the creation of gene cassettes with antibiotic resistance and phenotypic markers</td></tr><br />
<br />
<tr><td><a name="LacI-lacO-pLac-attI"></a>LacI-lacO-pLac-attI </td><td style="word-wrap: break-word; font-size: 8px;">GGGAATTCAAATCTAGAGACACCATCGAATGGTGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAATGAAGGAGGTGAATATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTCAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTAATAAGCGCAAAAAAAAAACCCCGCCCCTGACAGGGCGGGGTTTTTTTTCAACGCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAGGCAAAACTAGTAAACTGCAGGG</td><td>1403 bp</td><td>To be used as an AttI site with a Lac operon to allow controllable expression</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Template:Team:USyd-Australia/Template:StyleTemplate:Team:USyd-Australia/Template:Style2014-10-18T02:41:03Z<p>Andy.Bachler: </p>
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</html></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:40:23Z<p>Andy.Bachler: </p>
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{{:Team:USyd-Australia/Template:Banner}}<br />
<br />
<html><br />
<br />
<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template.</p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table id="gBlockTable" style="table-layout: fixed; width: 100%"><br />
<br />
<tr><th width="10%">Name</th><th width="45%">Sequence (5'-3')</th><th width="5%">Size</th><th width="20%">Purpose</th></tr><br />
<br />
<tr><td><a name="IntI1.2"></a>IntI1</td><td style="word-wrap: break-word; font-size: 8px;">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td>1121 bp</td><td>To be inserted into SamR construct as part of the controllable integrase expression system</td></tr><br />
<br />
<tr><td><a name="AttC-aeBlue-aacC1 "></a>AttC-aeBlue-aacC1 </td><td style="word-wrap: break-word; font-size: 8px;">TTTGAATTCGCGGCCGCTTCTAGAGTACCGAGGGACAGACTACCAACTCACAGCTAGCGCCTAACAATTCGTCCAAGCCGACGCCGCTTCGCGGCGCGGCTTAACTCAGGTGTTAGGCAAAGGAGGTTATTGATATGGCTTCACTGGTTAAAAAAGACATGTGCATCAAAATGACGATGGAAGGAACAGTAAACGGTCACCATTTCAAGTGTGTAGGAGAAGGCGAAGGCAAACCATTTGAAGGGACCCAGGTGGAAAAGATACGCATCACTGAAGGTGGGCCCTTACCATTTGCGTATGATATTTTGGCCCCTTGTTGCATGTATGGCAGTAAAACCTTCATTAAGCATGTGTCGGGTATTCCGGATTACTTTAAGGAGTCTTTTCCTGAGGGCTTTACCTGGGAAAGAACACAAATCTTCGAGGATGGCGGCTATCTCACCATACACCAGGACACGAGCCTTCAGGGTAATAATTTTATTTTCAAAGTTAATGTCATCGGTGCCAACTTCCCTGCAAACGGTCCCGTGATGCAGAAAAAAACAGCTGGATGGGAACCGTGCGTTGAGATGCTTTATCCGCGGGACGGCGTCCTGTGTGGTCAGAGCCTGATGGCCCTGAAATGCACTGATGGCAATCATCTGACGTCCCACCTGCGCACTACCTATCGTTCTCGCAAGCCATCCAATGCAGTTAACATGCCGGAATTTCATTTTGGGGATCATCGCATTGAGATTTTGAAAGCTGAACAAGGTAAATTTTATGAACAATACGAGTCAGCGGTGGCCCGTTACTGTGAGGCGGCACCGAGTAAATTAGGGCATCACTAATAAAGGATCCAAAGGAGGCTCAAGTATGGGCATCATTCGCACATGTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGATCTATATCTATGATCTCGCAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCGCGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGGAAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAATAATACTAGTAGCGGCCGCTGCAGCCC</td><td>1347 bp</td><td>To be used as a template for the creation of gene cassettes with antibiotic resistance and phenotypic markers</td></tr><br />
<br />
<tr><td><a name="LacI-lacO-pLac-attI"></a>LacI-lacO-pLac-attI </td><td style="word-wrap: break-word; font-size: 8px;">GGGAATTCAAATCTAGAGACACCATCGAATGGTGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAATGAAGGAGGTGAATATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTCAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTAATAAGCGCAAAAAAAAAACCCCGCCCCTGACAGGGCGGGGTTTTTTTTCAACGCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAGGCAAAACTAGTAAACTGCAGGG</td><td>1403 bp</td><td>To be used as an AttI site with a Lac operon to allow controllable expression</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:38:11Z<p>Andy.Bachler: </p>
<hr />
<div>{{Team:USyd-Australia/Template:Style}}<br />
{{:Team:USyd-Australia/Template:Banner}}<br />
<br />
<html><br />
<br />
<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template.</p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table borders="1px" 1id="gBlockTable" style="table-layout: fixed; width: 100%"><br />
<br />
<tr><th width="10%">Name</th><th width="45%">Sequence (5'-3')</th><th width="5%">Size</th><th width="20%">Purpose</th></tr><br />
<br />
<tr><td><a name="IntI1.2"></a>IntI1</td><td style="word-wrap: break-word; font-size: 8px;">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td>1121 bp</td><td>To be inserted into SamR construct as part of the controllable integrase expression system</td></tr><br />
<br />
<tr><td><a name="AttC-aeBlue-aacC1 "></a>AttC-aeBlue-aacC1 </td><td style="word-wrap: break-word; font-size: 8px;">TTTGAATTCGCGGCCGCTTCTAGAGTACCGAGGGACAGACTACCAACTCACAGCTAGCGCCTAACAATTCGTCCAAGCCGACGCCGCTTCGCGGCGCGGCTTAACTCAGGTGTTAGGCAAAGGAGGTTATTGATATGGCTTCACTGGTTAAAAAAGACATGTGCATCAAAATGACGATGGAAGGAACAGTAAACGGTCACCATTTCAAGTGTGTAGGAGAAGGCGAAGGCAAACCATTTGAAGGGACCCAGGTGGAAAAGATACGCATCACTGAAGGTGGGCCCTTACCATTTGCGTATGATATTTTGGCCCCTTGTTGCATGTATGGCAGTAAAACCTTCATTAAGCATGTGTCGGGTATTCCGGATTACTTTAAGGAGTCTTTTCCTGAGGGCTTTACCTGGGAAAGAACACAAATCTTCGAGGATGGCGGCTATCTCACCATACACCAGGACACGAGCCTTCAGGGTAATAATTTTATTTTCAAAGTTAATGTCATCGGTGCCAACTTCCCTGCAAACGGTCCCGTGATGCAGAAAAAAACAGCTGGATGGGAACCGTGCGTTGAGATGCTTTATCCGCGGGACGGCGTCCTGTGTGGTCAGAGCCTGATGGCCCTGAAATGCACTGATGGCAATCATCTGACGTCCCACCTGCGCACTACCTATCGTTCTCGCAAGCCATCCAATGCAGTTAACATGCCGGAATTTCATTTTGGGGATCATCGCATTGAGATTTTGAAAGCTGAACAAGGTAAATTTTATGAACAATACGAGTCAGCGGTGGCCCGTTACTGTGAGGCGGCACCGAGTAAATTAGGGCATCACTAATAAAGGATCCAAAGGAGGCTCAAGTATGGGCATCATTCGCACATGTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGATCTATATCTATGATCTCGCAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCGCGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGGAAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAATAATACTAGTAGCGGCCGCTGCAGCCC</td><td>1347 bp</td><td>To be used as a template for the creation of gene cassettes with antibiotic resistance and phenotypic markers</td></tr><br />
<br />
<tr><td><a name="LacI-lacO-pLac-attI"></a>LacI-lacO-pLac-attI </td><td style="word-wrap: break-word; font-size: 8px;">GGGAATTCAAATCTAGAGACACCATCGAATGGTGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAATGAAGGAGGTGAATATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTCAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTAATAAGCGCAAAAAAAAAACCCCGCCCCTGACAGGGCGGGGTTTTTTTTCAACGCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAGGCAAAACTAGTAAACTGCAGGG</td><td>1403 bp</td><td>To be used as an AttI site with a Lac operon to allow controllable expression</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:37:30Z<p>Andy.Bachler: </p>
<hr />
<div>{{Team:USyd-Australia/Template:Style}}<br />
{{:Team:USyd-Australia/Template:Banner}}<br />
<br />
<html><br />
<br />
<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template.</p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table borders="1px" 1id="gBlockTable" style="table-layout: fixed; width: 100%"><br />
<br />
<tr><th width="10%">Name</th><th width="40%">Sequence (5'-3')</th><th width="10%">Size</th><th width="20%">Purpose</th></tr><br />
<br />
<tr><td><a name="IntI1.2"></a>IntI1</td><td style="word-wrap: break-word; font-size: 6px;">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td>1121 bp</td><td>To be inserted into SamR construct as part of the controllable integrase expression system</td></tr><br />
<br />
<tr><td><a name="AttC-aeBlue-aacC1 "></a>AttC-aeBlue-aacC1 </td><td style="word-wrap: break-word; font-size: 6px;">TTTGAATTCGCGGCCGCTTCTAGAGTACCGAGGGACAGACTACCAACTCACAGCTAGCGCCTAACAATTCGTCCAAGCCGACGCCGCTTCGCGGCGCGGCTTAACTCAGGTGTTAGGCAAAGGAGGTTATTGATATGGCTTCACTGGTTAAAAAAGACATGTGCATCAAAATGACGATGGAAGGAACAGTAAACGGTCACCATTTCAAGTGTGTAGGAGAAGGCGAAGGCAAACCATTTGAAGGGACCCAGGTGGAAAAGATACGCATCACTGAAGGTGGGCCCTTACCATTTGCGTATGATATTTTGGCCCCTTGTTGCATGTATGGCAGTAAAACCTTCATTAAGCATGTGTCGGGTATTCCGGATTACTTTAAGGAGTCTTTTCCTGAGGGCTTTACCTGGGAAAGAACACAAATCTTCGAGGATGGCGGCTATCTCACCATACACCAGGACACGAGCCTTCAGGGTAATAATTTTATTTTCAAAGTTAATGTCATCGGTGCCAACTTCCCTGCAAACGGTCCCGTGATGCAGAAAAAAACAGCTGGATGGGAACCGTGCGTTGAGATGCTTTATCCGCGGGACGGCGTCCTGTGTGGTCAGAGCCTGATGGCCCTGAAATGCACTGATGGCAATCATCTGACGTCCCACCTGCGCACTACCTATCGTTCTCGCAAGCCATCCAATGCAGTTAACATGCCGGAATTTCATTTTGGGGATCATCGCATTGAGATTTTGAAAGCTGAACAAGGTAAATTTTATGAACAATACGAGTCAGCGGTGGCCCGTTACTGTGAGGCGGCACCGAGTAAATTAGGGCATCACTAATAAAGGATCCAAAGGAGGCTCAAGTATGGGCATCATTCGCACATGTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGATCTATATCTATGATCTCGCAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCGCGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGGAAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAATAATACTAGTAGCGGCCGCTGCAGCCC</td><td>1347 bp</td><td>To be used as a template for the creation of gene cassettes with antibiotic resistance and phenotypic markers</td></tr><br />
<br />
<tr><td><a name="LacI-lacO-pLac-attI"></a>LacI-lacO-pLac-attI </td><td style="word-wrap: break-word; font-size: 6px;">GGGAATTCAAATCTAGAGACACCATCGAATGGTGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAATGAAGGAGGTGAATATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTCAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTAATAAGCGCAAAAAAAAAACCCCGCCCCTGACAGGGCGGGGTTTTTTTTCAACGCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAGGCAAAACTAGTAAACTGCAGGG</td><td>1403 bp</td><td>To be used as an AttI site with a Lac operon to allow controllable expression</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:36:45Z<p>Andy.Bachler: </p>
<hr />
<div>{{Team:USyd-Australia/Template:Style}}<br />
{{:Team:USyd-Australia/Template:Banner}}<br />
<br />
<html><br />
<br />
<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template.</p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table borders="1px" 1id="gBlockTable" style="table-layout: fixed; width: 100%"><br />
<br />
<tr><th width="10%">Name</th><th width="60%">Sequence (5'-3')</th><th width="10%">Size</th><th width="20%">Purpose</th></tr><br />
<br />
<tr><td><a name="IntI1.2"></a>IntI1</td><td style="word-wrap: break-word; font-size: 10px;">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td>1121 bp</td><td>To be inserted into SamR construct as part of the controllable integrase expression system</td></tr><br />
<br />
<tr><td><a name="AttC-aeBlue-aacC1 "></a>AttC-aeBlue-aacC1 </td><td style="word-wrap: break-word; font-size: 10px;">TTTGAATTCGCGGCCGCTTCTAGAGTACCGAGGGACAGACTACCAACTCACAGCTAGCGCCTAACAATTCGTCCAAGCCGACGCCGCTTCGCGGCGCGGCTTAACTCAGGTGTTAGGCAAAGGAGGTTATTGATATGGCTTCACTGGTTAAAAAAGACATGTGCATCAAAATGACGATGGAAGGAACAGTAAACGGTCACCATTTCAAGTGTGTAGGAGAAGGCGAAGGCAAACCATTTGAAGGGACCCAGGTGGAAAAGATACGCATCACTGAAGGTGGGCCCTTACCATTTGCGTATGATATTTTGGCCCCTTGTTGCATGTATGGCAGTAAAACCTTCATTAAGCATGTGTCGGGTATTCCGGATTACTTTAAGGAGTCTTTTCCTGAGGGCTTTACCTGGGAAAGAACACAAATCTTCGAGGATGGCGGCTATCTCACCATACACCAGGACACGAGCCTTCAGGGTAATAATTTTATTTTCAAAGTTAATGTCATCGGTGCCAACTTCCCTGCAAACGGTCCCGTGATGCAGAAAAAAACAGCTGGATGGGAACCGTGCGTTGAGATGCTTTATCCGCGGGACGGCGTCCTGTGTGGTCAGAGCCTGATGGCCCTGAAATGCACTGATGGCAATCATCTGACGTCCCACCTGCGCACTACCTATCGTTCTCGCAAGCCATCCAATGCAGTTAACATGCCGGAATTTCATTTTGGGGATCATCGCATTGAGATTTTGAAAGCTGAACAAGGTAAATTTTATGAACAATACGAGTCAGCGGTGGCCCGTTACTGTGAGGCGGCACCGAGTAAATTAGGGCATCACTAATAAAGGATCCAAAGGAGGCTCAAGTATGGGCATCATTCGCACATGTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGATCTATATCTATGATCTCGCAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCGCGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGGAAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAATAATACTAGTAGCGGCCGCTGCAGCCC</td><td>1347 bp</td><td>To be used as a template for the creation of gene cassettes with antibiotic resistance and phenotypic markers</td></tr><br />
<br />
<tr><td><a name="LacI-lacO-pLac-attI"></a>LacI-lacO-pLac-attI </td><td style="word-wrap: break-word; font-size: 10px;">GGGAATTCAAATCTAGAGACACCATCGAATGGTGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAATGAAGGAGGTGAATATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTCAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTAATAAGCGCAAAAAAAAAACCCCGCCCCTGACAGGGCGGGGTTTTTTTTCAACGCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAGGCAAAACTAGTAAACTGCAGGG</td><td>To be used as an AttI site with a Lac operon to allow controllable expression</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:32:52Z<p>Andy.Bachler: </p>
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<div>{{Team:USyd-Australia/Template:Style}}<br />
{{:Team:USyd-Australia/Template:Banner}}<br />
<br />
<html><br />
<br />
<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template.</p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table borders="1px" 1id="gBlockTable" style="table-layout: fixed; width: 100%"><br />
<br />
<tr><th width="20%">Name</th><th width="60%">Sequence (5'-3')</th><th width="20%">Purpose</th></tr><br />
<br />
<tr><td><a name="IntI1.2"></a>IntI1</td><td style="word-wrap: break-word; font-size: 10px;">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td>To be inserted into SamR construct as part of the controllable integrase expression system</td></tr><br />
<br />
<tr><td><a name="AttC-aeBlue-aacC1 "></a>AttC-aeBlue-aacC1 </td><td style="word-wrap: break-word; font-size: 10px;">TTTGAATTCGCGGCCGCTTCTAGAGTACCGAGGGACAGACTACCAACTCACAGCTAGCGCCTAACAATTCGTCCAAGCCGACGCCGCTTCGCGGCGCGGCTTAACTCAGGTGTTAGGCAAAGGAGGTTATTGATATGGCTTCACTGGTTAAAAAAGACATGTGCATCAAAATGACGATGGAAGGAACAGTAAACGGTCACCATTTCAAGTGTGTAGGAGAAGGCGAAGGCAAACCATTTGAAGGGACCCAGGTGGAAAAGATACGCATCACTGAAGGTGGGCCCTTACCATTTGCGTATGATATTTTGGCCCCTTGTTGCATGTATGGCAGTAAAACCTTCATTAAGCATGTGTCGGGTATTCCGGATTACTTTAAGGAGTCTTTTCCTGAGGGCTTTACCTGGGAAAGAACACAAATCTTCGAGGATGGCGGCTATCTCACCATACACCAGGACACGAGCCTTCAGGGTAATAATTTTATTTTCAAAGTTAATGTCATCGGTGCCAACTTCCCTGCAAACGGTCCCGTGATGCAGAAAAAAACAGCTGGATGGGAACCGTGCGTTGAGATGCTTTATCCGCGGGACGGCGTCCTGTGTGGTCAGAGCCTGATGGCCCTGAAATGCACTGATGGCAATCATCTGACGTCCCACCTGCGCACTACCTATCGTTCTCGCAAGCCATCCAATGCAGTTAACATGCCGGAATTTCATTTTGGGGATCATCGCATTGAGATTTTGAAAGCTGAACAAGGTAAATTTTATGAACAATACGAGTCAGCGGTGGCCCGTTACTGTGAGGCGGCACCGAGTAAATTAGGGCATCACTAATAAAGGATCCAAAGGAGGCTCAAGTATGGGCATCATTCGCACATGTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGATCTATATCTATGATCTCGCAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCGCGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGGAAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAATAATACTAGTAGCGGCCGCTGCAGCCC</td><td>To be used as a template for the creation of gene cassettes with antibiotic resistance and phenotypic markers</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:32:20Z<p>Andy.Bachler: </p>
<hr />
<div>{{Team:USyd-Australia/Template:Style}}<br />
{{:Team:USyd-Australia/Template:Banner}}<br />
<br />
<html><br />
<br />
<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template.</p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table borders="1px" 1id="gBlockTable" style="table-layout: fixed; width: 100%"><br />
<br />
<tr><th width="20%">Name</th><th width="60%">Sequence (5'-3')</th><th width="20%">Purpose</th></tr><br />
<br />
<tr><td><a name="IntI1.2"></a>IntI1</td><td style="word-wrap: break-word">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td>To be inserted into SamR construct as part of the controllable integrase expression system</td></tr><br />
<br />
<tr><td><a name="AttC-aeBlue-aacC1 "></a>AttC-aeBlue-aacC1 </td><td style="word-wrap: break-word">TTTGAATTCGCGGCCGCTTCTAGAGTACCGAGGGACAGACTACCAACTCACAGCTAGCGCCTAACAATTCGTCCAAGCCGACGCCGCTTCGCGGCGCGGCTTAACTCAGGTGTTAGGCAAAGGAGGTTATTGATATGGCTTCACTGGTTAAAAAAGACATGTGCATCAAAATGACGATGGAAGGAACAGTAAACGGTCACCATTTCAAGTGTGTAGGAGAAGGCGAAGGCAAACCATTTGAAGGGACCCAGGTGGAAAAGATACGCATCACTGAAGGTGGGCCCTTACCATTTGCGTATGATATTTTGGCCCCTTGTTGCATGTATGGCAGTAAAACCTTCATTAAGCATGTGTCGGGTATTCCGGATTACTTTAAGGAGTCTTTTCCTGAGGGCTTTACCTGGGAAAGAACACAAATCTTCGAGGATGGCGGCTATCTCACCATACACCAGGACACGAGCCTTCAGGGTAATAATTTTATTTTCAAAGTTAATGTCATCGGTGCCAACTTCCCTGCAAACGGTCCCGTGATGCAGAAAAAAACAGCTGGATGGGAACCGTGCGTTGAGATGCTTTATCCGCGGGACGGCGTCCTGTGTGGTCAGAGCCTGATGGCCCTGAAATGCACTGATGGCAATCATCTGACGTCCCACCTGCGCACTACCTATCGTTCTCGCAAGCCATCCAATGCAGTTAACATGCCGGAATTTCATTTTGGGGATCATCGCATTGAGATTTTGAAAGCTGAACAAGGTAAATTTTATGAACAATACGAGTCAGCGGTGGCCCGTTACTGTGAGGCGGCACCGAGTAAATTAGGGCATCACTAATAAAGGATCCAAAGGAGGCTCAAGTATGGGCATCATTCGCACATGTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGATCTATATCTATGATCTCGCAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCGCGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGGAAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAATAATACTAGTAGCGGCCGCTGCAGCCC</td><td>To be used as a template for the creation of gene cassettes with antibiotic resistance and phenotypic markers</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:29:53Z<p>Andy.Bachler: </p>
<hr />
<div>{{Team:USyd-Australia/Template:Style}}<br />
{{:Team:USyd-Australia/Template:Banner}}<br />
<br />
<html><br />
<br />
<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template. 3 were reconstituted in 10&#956;L TE buffer </p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table borders="1px" 1id="gBlockTable" style="table-layout: fixed; width: 100%"><br />
<br />
<tr><th width="20%">Name</th><th width="60%">Sequence (5'-3')</th><th width="20%">Purpose</th></tr><br />
<tr><td><a name="IntI1.2"></a>IntI1</td><td style="word-wrap: break-word">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td>To be inserted into SamR construct as part of the controllable integrase expression system</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:25:24Z<p>Andy.Bachler: </p>
<hr />
<div>{{Team:USyd-Australia/Template:Style}}<br />
{{:Team:USyd-Australia/Template:Banner}}<br />
<br />
<html><br />
<br />
<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template. 3 were reconstituted in 10&#956;L TE buffer </p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table id="gBlockTable" style="table-layout: fixed; width: 100%"><br />
<br />
<tr><th width="20%">Name</th><th width="60%">Sequence (5'-3')</th><th width="20%">Purpose</th></tr><br />
<tr><td><a name="IntI1.2"></a>IntI1</td><td style="word-wrap: break-word">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td>To be inserted into SamR construct as part of the controllable integrase expression system</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:23:54Z<p>Andy.Bachler: </p>
<hr />
<div>{{Team:USyd-Australia/Template:Style}}<br />
{{:Team:USyd-Australia/Template:Banner}}<br />
<br />
<html><br />
<br />
<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template. 3 were reconstituted in 10&#956;L TE buffer </p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table id="gBlockTable" style="table-layout: fixed; width: 100%"><br />
<br />
<tr><th >Name</th><th>Sequence (5'-3')</th><th>Purpose</th></tr><br />
<tr><td><a name="IntI1.2"></a>IntI1</td><td style="word-wrap: break-word">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td>To be inserted into SamR construct as part of the controllable integrase expression system</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:23:00Z<p>Andy.Bachler: </p>
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<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template. 3 were reconstituted in 10&#956;L TE buffer </p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table id="gBlockTable" style="table-layout: fixed; width: 100%"><br />
<br />
<tr><th width="10%">Name</th><th>Sequence (5'-3')</th><th>Purpose</th></tr><br />
<tr><td><a name="IntI1.2"></a>IntI1</td><td style="word-wrap: break-word">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td>58.2C</td><td>Insert into SamR construct to be used as part of the controllable integrase expression system</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Template:Team:USyd-Australia/Template:StyleTemplate:Team:USyd-Australia/Template:Style2014-10-18T02:22:56Z<p>Andy.Bachler: </p>
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</html></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:21:58Z<p>Andy.Bachler: </p>
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<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template. 3 were reconstituted in 10&#956;L TE buffer </p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table id="gBlockTable" style="table-layout: fixed; width: 100%"><br />
<br />
<tr><th width="10%">Name</th><th>Sequence (5'-3')</th>><th>Purpose</th></tr><br />
<tr><td><a name="IntI1.2"></a>IntI1.2</td><td style="word-wrap: break-word">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td>58.2C</td><td>Insert into SamR construct to be used as part of the controllable integrase expression system</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:21:39Z<p>Andy.Bachler: </p>
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<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template. 3 were reconstituted in 10&#956;L TE buffer </p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table id="gBlockTable" style="table-layout: fixed; width: 100%"><br />
<br />
<tr><th width="10%">Name</th><th style="max-width:500px;">Sequence (5'-3')</th><th width="10%">Theoretical Tm</th><th>Purpose</th></tr><br />
<tr><td><a name="IntI1.2"></a>IntI1.2</td><td style="word-wrap: break-word">CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td>58.2C</td><td>Insert into SamR construct to be used as part of the controllable integrase expression system</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/Notebook/gBlockTeam:USyd-Australia/Notebook/gBlock2014-10-18T02:21:02Z<p>Andy.Bachler: </p>
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<table width="90%" align="center"><br />
<tr><td> <h2>gBlocks</h2></td></tr><br />
<tr><td><br />
<p>All gBlocks were supplied by IDT.</p><br />
<p>gBlocks are DNA chunks up to 2kb made from a sequence template. 3 were reconstituted in 10&#956;L TE buffer </p><br />
<p></p><br />
</td></tr><br />
<br />
</table><br />
<table id="gBlockTable" align="center"><br />
<br />
<tr><th width="10%">Name</th><th style="max-width:500px;">Sequence (5'-3')</th><th width="10%">Theoretical Tm</th><th>Purpose</th></tr><br />
<tr><td><a name="IntI1.2"></a>IntI1.2</td><td>CCGTTTTTTTGGATGGAGTGAAACGATGGCGATTGCAATAGCTAGCTAAAGGAGGATTCATATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAGGTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACTCCCTCCGAACCGAACAGGCTTACGTTAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGGCAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGCAAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGCAAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGAAGACCTCGGCCGTCGCGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTGGAAGGCGAGCATCGTTTGTTCGCCCAGCTTCTGTATGGAACGGGCATGCGGATCAGTGAGGGTTTGCAACTGCGGGTCAAGGATCTGGATTTCGATCACGGCACGATCATCGTGCGGGAGGGCAAGGGCTCCAAGGATCGGGCCTTGATGTTACCCGAGAGCTTGGCACCCAGCCTGCGCGAGCAGCTGTCGCGTGCACGGGCATGGTGGCTGAAGGACCAGGCCGAGGGCCGCAGCGGCGTTGCGCTTCCCGACGCCCTTGAGCGGAAGTATCCGCGCGCCGGGCATTCCTGGCCGTGGTTCTGGGTTTTTGCGCAGCACACGCATTCGACCGATCCACGGAGCGGTGTCGTGCGTCGCCATCACATGTATGACCAGACCTTTCAGCGCGCCTTCAAACGTGCCGTAGAACAAGCAGGCATCACGAAGCCCGCCACACCGCACACCCTCCGCCACTCGTTCGCGACGGCCTTGCTCCGCAGCGGTTACGACATTCGAACCGTGCAGGATCTGCTCGGCCATTCCGACGTCTCTACGACGATGATTTACACGCATGTGCTGAAAGTTGGCGGTGCCGGAGTGCGCTCACCGCTTGATGCGCTGCCGCCCCTCACTTCGGAGAGGTAGTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCA</td><td>58.2C</td><td>Insert into SamR construct to be used as part of the controllable integrase expression system</td></tr><br />
</table><br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/pUS203Team:USyd-Australia/pUS2032014-10-17T23:34:04Z<p>Andy.Bachler: </p>
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<h2><a href="http://parts.igem.org/Part:BBa_K1388000">pUS203 Controllable Integrase production</a></h2><br />
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<p style="font-size:12px"><br />
To see this part on the iGEM Parts catalogue please click <a href="http://parts.igem.org/Part:BBa_K1388000">here</a>. <br>Sequences for this part are also available in <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLRDJ3OVdUUlhEVjA/view?usp=sharing">SnapGene (.dna file)</a>, <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLT3hQZno4VXFVZ0U/view?usp=sharing">FASTA (.fa file)</a>, <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLbndkclF5UlFNVE0/view?usp=sharing">GenBank Standard (.gb file)</a> and <a href="https://drive.google.com/file/d/0B4B2M7gt-JMLak5Qdi1CbFZ4bm8/view?usp=sharing">Plain Text (.txt file)</a> file format.<br />
</p><br />
<h2>Aim</h2><br />
<p>Our aim was to create a controllable Integrase producing plasmid as a submission part. From the submission plasmid the controllable Integrase Part could then be ligated into a convenient low-copy plasmid to help reduce the potential toxicity of Integrase to the cell.</p><br />
<br />
<h2>Approach</h2><br />
<p><br />
<img src="https://static.igem.org/mediawiki/2014/6/67/PUS203.png" align="right" width="50%"><br />
The 2010 Paris Bettencourt team had developed a <a href=http://parts.igem.org/wiki/index.php?title=Part:BBa_K329013>controllable Integrase part</a> for use a population counter in their project. We contacted them to see if we could source the part from them but it was unfortunately not possible. We set about creating the BioBrick using the araC-pBAD from the well characterized part <a href=http://parts.igem.org/Part:BBa_K731201>K731201</a> and ordering the IntI1 as a gBlock. The araC-pBAD system in pSB1C3 had previously been prepared by Honours student Sam Ross (called the SamR construct). Our next steps were the design of the gBlock and integration of the IntI1 gBlock into the SamR construct.<br />
</p><br />
<br />
<h2>Materials</h2><br />
<ul>DNA<ul><br />
<li>SamR Construct: araC-pBAD in pSC1B3</li><br />
<li><a href="#">IntI1 gBlock</a>: to be inserted into SamR</li><br />
<li>pUS44: Verified plasmid containing an AttI site, used in Validation</li><br />
<li>aeBlue: Cassette PCR'ed from <a href=http://parts.igem.org/Part:BBa_K1033929>K1033929</a> for us in validation</li><br />
</ul><br />
</ul><br />
<br />
<ul>Primers<br />
<table><br />
<tr><br />
<td width="50%"><br />
<ul><br />
<li><a href=”#”>iGEM1413</a></li><br />
<li><a href=”#”>iGEM1414</a></li><br />
<li><a href=”#”>iGEM1416</a></li></ul><br />
</td><br />
<td><br />
<ul><br />
<li><a href=”#”>iGEM16</a></li><br />
<li><a href=”#”>iGEM25</a></li><br />
</ul><br />
</td><br />
</tr><br />
<tr><br />
</table><br />
<br />
<h2>Inti1 gBlock Design</h2><br />
<p><br />
The code for the sequence for the IntI1 gene was sourced from pUS2056 but is highly conserved so this choice was out of convenience as the sequence was locally available. It was modified by Tom Geddes, Rokiah Alford and our supervisor Dr Nicholas Coleman to:<br><ul><br />
<li>Include a Sac1 restriction enzyme site at the beginning before the Inti1 gene so it could be excised independently of the promoter if needed</li><br />
<li>Include a different ribosome binding site </li><br />
<li>Include Gibson ends that overlapped with the backbone for simple Gibson Assembly later with the SamR construct.</li><br />
<li>Remove an illegal Spe1 site near the end to make it compliant with iGEM requirements</li><br />
<li>Remove a promoter pC that was coded in the opposite direction to the promoter we wanted to use and would have clashed with the controllable promotion of this part</li></ul><br><br />
Our first attempt to Gibson Assemble the SamR construct and IntI1 gBlock failed because there was extra code after the code for pBAD which we had not anticipated. This meant that the Gibson Assembly could not properly integrate the gBlock. We discussed numerous options to try and either get rid of the problem sequence on the SamR construct or modify the ends of the gBlock to accommodate the problem sequence. In the end it was decided that a re-design of the gBlock would be the most efficient way to solve the problem. <br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<h2>Inti1 gBlock Design</h2><br />
<p><br />
Three major changes were decided: <br><br />
<ul><br />
<li>Changed the code for the ribosome cut site as a base pair was incorrect</li><br />
<li>To put an Nhe1 restriction enzyme cut site between the IntI1 gene and the promoters on the SamR construct. This was done to help in the ligation of this BioBrick into the low-copy backbone pUS202. </li><br />
<li>Changing of the end sequences to accommodate the extra code present on the SamR construct</li><br />
</ul><br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<br />
<h2>Construction</h2><br />
<p><br />
The IntI1 gBlock was inserted into the SamR construct using <a href="http://www.addgene.org/plasmid-protocols/gibson-assembly/">Gibson Asssembly</a>. The ends of the gBlock in the redesign were compatible with the ends of the SamR construct and after running the Gibson Assembly reaction we used a variety of methods to prove that our gBlock had inserted correctly.<br />
</p><br />
<br />
<p><br />
These changed were performed by our supervisor Nick Coleman in consultation with Rokiah Alford. The redesigned gBlock was able to be successfully Gibsoned into the SamR backbone. To verify the presence of the insert we used junction primers which would amplify over both ends of the insert and could only amplify if the gBlock had been integrated into the SamR construct. <br><br />
<br><b>Junction primer PCR</b><br><br />
<br />
Junction primers for the junction of promoter and gblock were iGEM1413 and iGEM1414 and were expected to give a fragment size of 377 base pairs. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/4/4e/USyd-AustraliaWell_Images_restriction_Digest.png"> <br><br />
<br />
9 out of the 12 colonies screened gave a positive result and these were further screened with a second set of junction primers on the other side of the gBlock. The junction primers for this were iGEM1416 and iGEM25 and were expected to yielad a fragment size of 432bp. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/7/70/USyd-Australia_RealSecondPCR_Junction_screen.png"> <br><br />
<br />
Again 9 of the 12 colonies were positive here. The PCR product was sequenced and corresponded with the expected sequence for the plasmid. 3 of the 8 colonies were selected to be grown up and Plasmid Prepped to remove the plasmid and allow restriction digest and sequencing of the plasmid for further verification.<br><br />
<br />
<br><b>Restriction Digest</b><br><br />
<br />
The pUS203 plasmid was further confirmed with restriction digestion. Spe1, Pst1 and EcoR1 are in the prefix and suffix and should cut. An Nhe1 restriction site was added into the gBlock code and so a cut with this enzyme would indicate the gBlock had been inserted. Sac1 should not cut and should be identical to the uncut control lane. <br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/4/4e/USyd-AustraliaWell_Images_restriction_Digest.png"><br><br />
<br />
The digest results were as expected using SnapGene’s Virtual Digest tool. We were satisfied with these results despite the Sac1 error because the other fragments were as expected and most importantly because Nhe1 cut, which could only have occurred if the gblock had been incorporated into the plasmid. The Sac1 sample was heat-treated to inactive the Sac1 enzyme and used was used as a template to PCR the BioBrick. The results were still as expected which showed that the unexpected action of the Sac1 seen in the gel for the digest of this plasmid luckily didn’t reflect an error in the biobrick code. <br />
<b>Biobrick PCR and sequencing</b><br />
The final check for this plasmid was to PCR the entire biobrick fragment using iGEM16 and iGEM25 and sequencing the product. <br />
</p><br />
<br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
<h2>Validation</h2><br />
<ul style="font-weight: normal;"><br />
To validate our araC-pBAD/IntI1 BioBrick it would need to be present in a low copy plasmid in order to reduce the amount produced as it is potentially toxic to the cell at high concentrations. Our intial plan was to insert the BioBrick into pUS202 but it turned out not to be low-copy after validation. The back up option was to ligate the BioBrick into pSB6A1. The initial attempt was not successful and due to time constraints we decided to try and test the integron construct in the pSB1C3 it was currently in even with the potential toxicity.<br><br><br />
To test for the proper expression of our integron gene in the BioBrick we wanted to have an AttI site and integrase protein present in E. coli. This was so that upon transformation with a cassette (which has an AttC site) a co-integrate would quickly form with the AttI site and before the cassettes could be lost out of the cell. Upon integration of the cassette the Gentomycin resistance (GmR) present on the cassette is expressed allowing us to select for the correct cells on Gentomycin plates. The negative control had no arabinose induction and so the cassette should not integrate and the cells should not be Gm resistant.<br><br><br />
<img src="https://static.igem.org/mediawiki/2014/7/71/USyd-Australia_Integrase_diagrams_1.png" width="98%"><br><br><br />
The initial experiments did not yield positive results and due to time constraints we have not been able to troubleshoot the experiment. <br />
</ul><br />
</li><br />
<a href="#protstart">Back to top</a><!------------------------------------------------------------------------------------------------------------><br />
<br><br><br />
<br />
</html></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/OutreachTeam:USyd-Australia/Outreach2014-10-17T23:33:45Z<p>Andy.Bachler: </p>
<hr />
<div>{{Team:USyd-Australia/Template:Style}}<br />
{{:Team:USyd-Australia/Template:Banner}}<br />
<br />
<html><br />
<br />
<div><br />
<br />
<h1>Our Goals</h1><br />
<br />
<a href="http://strangenature.org/"><img src="https://static.igem.org/mediawiki/2014/a/a8/USyd-Australia_SN-Poster.jpg" align="right" width="25%" hspace="10px" vspace="40px"></a><br />
Our project is in the “Foundational Advance” track and similarly we wanted High School students to learn and investigate what Synthetic Biology is, and where it is heading in the future. In 2013, the University of Sydney iGEM team created <a href="http://2013.strangenature.org/"> Strange Nature</a>, a Synthetic Biology writing competition for high school students, asking the question "What problems will be caused or solved by synthetic biology?". We provided students with inspiration through our online learning platform, giving them a multitude of examples of the cool and kooky ways that synthetic biology can be used.<br><br><br />
<br />
We believe in the saying 'to create is to learn', so this year we decided to continue the writing competition, asking for students to share their opinions about synthetic biology in 1000 words. We asked students to create a 1000 word piece answering the following question: <br><br><br />
<br />
<h3 align="center"> “Which advancement in synthetic biology do you think <br><br />
is the most promising, and why?”</h3><br><br />
<br />
While genetic engineering is superficially covered in the NSW and Australian High School curriculum we felt it important to try and get students to think beyond what they've been taught in class. Our aim was to encourage students to find information on Synthetic Biology on their own, and to raise awareness of the field.<br><br><br />
<br />
<h1>Competition</h1><br />
<br />
Our competition was hosted on the <a href="http://www.StrangeNature.org">Strange Nature</a> website where we provided resources about what Synthetic Biology was, recent developments and future applications. The competition was open until the 1st October. We had a whole host of submissions from Dystopian future stories of genetically engineered super humans to exploding <i>E. coli</i>. We had more essays submitted this year than previously and while the quality of submissions varied they were all incredibly insightful to what the students think about the possibilities of Synthetic Biology. We currently have a short list of 6 pieces which we feel are well written, explore Synthetic Biology and show imagination in their responses.<br><br><br />
<br />
<h1>Out and About</h1><br />
<br />
<h4>University Open Days</h4><br />
<br />
Over the course of the year we volunteered at our Universities Open Days, spruiking both studying Science and participating in the Strange Nature writing competition. Talking to future science students about what is possible and what may soon be possible was incredibly insightful and seeing their interest in Synthetic Biology was wonderful.<br><br><br />
<img src="https://static.igem.org/mediawiki/2014/e/e9/USyd-Australia_Kya_Explaining_Things.png" width="400px" height="200px"hspace="100"> <br />
<img src="https://static.igem.org/mediawiki/2014/0/03/USyd-Australia_Callum_Holding_Bacteria.png" width="300px" height="300px"><br />
<br><br />
<br />
<p style="font-size: 80%;"><br />
Pictured above are Kya (left) and Callum (right) talking to prospective Science students about everything from Genetic Engineering to the best place for coffee on campus.</p><br />
<br />
<h4>Science Week Volunteering</h4><br />
<br />
We also volunteered as part of the Australian Society for Microbiology during Science Week at their stall at the Australian Museum. There we were able to talk to hundreds of Primary (years K-6), High School students (years 7-12), and general members of the public to discuss Microbiology, Biotechnology and also find out what they knew about Synthetic Biology and genetic engineering. The displays really fascinated the kids and students, especially the fluorescent bacteria! <br><br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/e/ef/USyd-Australia_Pretty_Plate_images.png" width="550px" height="200px" hspace="200px" alt="Fluorescent Bacteria in long wave UV light cupboard"><br />
<br />
<h4>Contacting Teachers and Schools</h4><br />
<br />
We got in contact with many teachers and schools to promote the writing competition. We also wrote articles for Science Resource Newsletters sent to teachers to try and increase the awareness of our project.<br><br><br />
<br />
<h1>Collaboration </h1><br />
<br />
<h4>University of Melbourne</h4><br />
<br />
In preparation for promoting the competition, we contacted the <a href="https://2014.igem.org/Team:Melbourne"> University of Melbourne </a> iGEM team to see if they would like to collaborate. As part of their outreach, they decided to create their own informative video about Synthetic Biology for high school students, including a one minute feature about Strange Nature!<br><br />
To see the video, follow this link <a href="https://www.youtube.com/watch?v=ZuSoO3HVMjo"> here </a><br />
<br />
<source src="https://static.igem.org/mediawiki/2014/8/8a/USyd-Australia_Strange_Nature_Video.mov" type="video/mov"><br />
<br />
<h1>Result</h1><br />
In total, 32 entries were received from all around Australia, ranging from as young as grade 8, up to year 12 students. The best pieces we received demonstrated they had done some research into what Synthetic Biology actually is and the possibilities this new field can bring. The vast majority of submissions we received were incredibly positive about the technology and in many submissions the level of excitement conveyed was incredibly encouraging. <br><br><br />
<br />
To go beyond simply giving a prize for the "best" piece we are providing feedback for each submission to correct some mistakes and also to start a discussion on their topics. We believe that this would help to create a healthy dialogue between interested members of the public, and the research scientists behind the technologies. Many pieces only considered the positive side of an advance, such as <i>in vitro</i> meat, while not addressing the negative problems. Our aim was to encourage students to consider the future of Synthetic Biology and through their submission and the feedback we provide it is our belief that we are prompting them to broaden their thinking of the advances which may come in the future. <br><br><br />
<br />
To see some of the submissions we received, pick a title below!<br><br><br />
<br />
<a href="https://static.igem.org/mediawiki/2014/8/86/USyd-Australia_SN_Submission_36_Suicide_E._coli.pdf">Suicide E. coli</a><br><br />
<a href="https://static.igem.org/mediawiki/2014/1/19/USyd-Australia_SN_Submission_32_SynthBioGeneral.pdf">Synthetic Biology Review</a><br><br />
<a href="https://static.igem.org/mediawiki/2014/9/95/USyd-Australia_SN_Submission_11_LethalGeneInMosquito.pdf">Lethal Gene for Mosquito Control</a><br><br />
<a href="https://static.igem.org/mediawiki/2014/d/d2/USyd-Australia_SN_Submission_7_Artemisinin.pdf">Artemisinin Production from Yeast</a><br />
<br />
<div> <br />
<br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/File:USyd-Australia_SN_Submission_7_Artemisinin.pdfFile:USyd-Australia SN Submission 7 Artemisinin.pdf2014-10-17T23:32:47Z<p>Andy.Bachler: </p>
<hr />
<div></div>Andy.Bachlerhttp://2014.igem.org/Team:USyd-Australia/OutreachTeam:USyd-Australia/Outreach2014-10-17T23:30:37Z<p>Andy.Bachler: </p>
<hr />
<div>{{Team:USyd-Australia/Template:Style}}<br />
{{:Team:USyd-Australia/Template:Banner}}<br />
<br />
<html><br />
<br />
<div><br />
<br />
<h1>Our Goals</h1><br />
<br />
<a href="http://strangenature.org/"><img src="https://static.igem.org/mediawiki/2014/a/a8/USyd-Australia_SN-Poster.jpg" align="right" width="25%" hspace="10px" vspace="40px"></a><br />
Our project is in the “Foundational Advance” track and similarly we wanted High School students to learn and investigate what Synthetic Biology is, and where it is heading in the future. In 2013, the University of Sydney iGEM team created <a href="http://2013.strangenature.org/"> Strange Nature</a>, a Synthetic Biology writing competition for high school students, asking the question "What problems will be caused or solved by synthetic biology?". We provided students with inspiration through our online learning platform, giving them a multitude of examples of the cool and kooky ways that synthetic biology can be used.<br><br><br />
<br />
We believe in the saying 'to create is to learn', so this year we decided to continue the writing competition, asking for students to share their opinions about synthetic biology in 1000 words. We asked students to create a 1000 word piece answering the following question: <br><br><br />
<br />
<h3 align="center"> “Which advancement in synthetic biology do you think <br><br />
is the most promising, and why?”</h3><br><br />
<br />
While genetic engineering is superficially covered in the NSW and Australian High School curriculum we felt it important to try and get students to think beyond what they've been taught in class. Our aim was to encourage students to find information on Synthetic Biology on their own, and to raise awareness of the field.<br><br><br />
<br />
<h1>Competition</h1><br />
<br />
Our competition was hosted on the <a href="http://www.StrangeNature.org">Strange Nature</a> website where we provided resources about what Synthetic Biology was, recent developments and future applications. The competition was open until the 1st October. We had a whole host of submissions from Dystopian future stories of genetically engineered super humans to exploding <i>E. coli</i>. We had more essays submitted this year than previously and while the quality of submissions varied they were all incredibly insightful to what the students think about the possibilities of Synthetic Biology. We currently have a short list of 6 pieces which we feel are well written, explore Synthetic Biology and show imagination in their responses.<br><br><br />
<br />
<h1>Out and About</h1><br />
<br />
<h4>University Open Days</h4><br />
<br />
Over the course of the year we volunteered at our Universities Open Days, spruiking both studying Science and participating in the Strange Nature writing competition. Talking to future science students about what is possible and what may soon be possible was incredibly insightful and seeing their interest in Synthetic Biology was wonderful.<br><br><br />
<img src="https://static.igem.org/mediawiki/2014/e/e9/USyd-Australia_Kya_Explaining_Things.png" width="400px" height="200px"hspace="100"> <br />
<img src="https://static.igem.org/mediawiki/2014/0/03/USyd-Australia_Callum_Holding_Bacteria.png" width="300px" height="300px"><br />
<br><br />
<br />
<p style="font-size: 80%;"><br />
Pictured above are Kya (left) and Callum (right) talking to prospective Science students about everything from Genetic Engineering to the best place for coffee on campus.</p><br />
<br />
<h4>Science Week Volunteering</h4><br />
<br />
We also volunteered as part of the Australian Society for Microbiology during Science Week at their stall at the Australian Museum. There we were able to talk to hundreds of Primary (years K-6), High School students (years 7-12), and general members of the public to discuss Microbiology, Biotechnology and also find out what they knew about Synthetic Biology and genetic engineering. The displays really fascinated the kids and students, especially the fluorescent bacteria! <br><br><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/e/ef/USyd-Australia_Pretty_Plate_images.png" width="550px" height="200px" hspace="200px" alt="Fluorescent Bacteria in long wave UV light cupboard"><br />
<br />
<h4>Contacting Teachers and Schools</h4><br />
<br />
We got in contact with many teachers and schools to promote the writing competition. We also wrote articles for Science Resource Newsletters sent to teachers to try and increase the awareness of our project.<br><br><br />
<br />
<h1>Collaboration </h1><br />
<br />
<h4>University of Melbourne</h4><br />
<br />
In preparation for promoting the competition, we contacted the <a href="https://2014.igem.org/Team:Melbourne"> University of Melbourne </a> iGEM team to see if they would like to collaborate. As part of their outreach, they decided to create their own informative video about Synthetic Biology for high school students, including a one minute feature about Strange Nature!<br><br />
To see the video, follow this link <a href="https://www.youtube.com/watch?v=ZuSoO3HVMjo"> here </a><br />
<br />
<source src="https://static.igem.org/mediawiki/2014/8/8a/USyd-Australia_Strange_Nature_Video.mov" type="video/mov"><br />
<br />
<h1>Result</h1><br />
In total, 32 entries were received from all around Australia, ranging from as young as grade 8, up to year 12 students. The best pieces we received demonstrated they had done some research into what Synthetic Biology actually is and the possibilities this new field can bring. The vast majority of submissions we received were incredibly positive about the technology and in many submissions the level of excitement conveyed was incredibly encouraging. <br><br><br />
<br />
To go beyond simply giving a prize for the "best" piece we are providing feedback for each submission to correct some mistakes and also to start a discussion on their topics. We believe that this would help to create a healthy dialogue between interested members of the public, and the research scientists behind the technologies. Many pieces only considered the positive side of an advance, such as <i>in vitro</i> meat, while not addressing the negative problems. Our aim was to encourage students to consider the future of Synthetic Biology and through their submission and the feedback we provide it is our belief that we are prompting them to broaden their thinking of the advances which may come in the future. <br><br><br />
<br />
To see some of the submissions we received, pick a title below!<br><br><br />
<br />
<a href="https://static.igem.org/mediawiki/2014/8/86/USyd-Australia_SN_Submission_36_Suicide_E._coli.pdf">Suicide E. coli</a><br><br />
<a href="https://static.igem.org/mediawiki/2014/1/19/USyd-Australia_SN_Submission_32_SynthBioGeneral.pdf">Synthetic Biology Review</a><br><br />
<a href="https://static.igem.org/mediawiki/2014/9/95/USyd-Australia_SN_Submission_11_LethalGeneInMosquito.pdf">Lethal Gene for Mosquito Control</a><br><br />
<br />
<div> <br />
<br />
</html><br />
{{Team:USyd-Australia/Footer}}</div>Andy.Bachlerhttp://2014.igem.org/File:USyd-Australia_SN_Submission_11_LethalGeneInMosquito.pdfFile:USyd-Australia SN Submission 11 LethalGeneInMosquito.pdf2014-10-17T23:28:45Z<p>Andy.Bachler: </p>
<hr />
<div></div>Andy.Bachlerhttp://2014.igem.org/File:USyd-Australia_SN_Submission_32_SynthBioGeneral.pdfFile:USyd-Australia SN Submission 32 SynthBioGeneral.pdf2014-10-17T23:26:54Z<p>Andy.Bachler: </p>
<hr />
<div></div>Andy.Bachlerhttp://2014.igem.org/File:USyd-Australia_SN_Submission_36_Suicide_E._coli.pdfFile:USyd-Australia SN Submission 36 Suicide E. coli.pdf2014-10-17T23:23:25Z<p>Andy.Bachler: </p>
<hr />
<div></div>Andy.Bachler