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− | <div class=" topimg pull-right sm-no-float col-md- | + | <div class=" topimg pull-right sm-no-float col-md-7 "> |
− | <img src="https://static.igem.org/mediawiki/2019/3/31/T--DTU-Denmark--partsoverview2.svg" | + | <img src="https://static.igem.org/mediawiki/2019/3/31/T--DTU-Denmark--partsoverview2.svg" alt="Illutration of a puzzle of fungal parts" style="margin-top:75px;max-width:70%;margin-right:auto; margin-left:auto;display: block; |
height:auto;z-index:5;"> | height:auto;z-index:5;"> | ||
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− | <div class="pull-left col-md- | + | <div class="pull-left col-md-5 sm-text-center com-sm-12 "> |
− | <div class="team-overview"> | + | <div class="team-overview" style="font-size:1em";> |
<h2>Parts overview</h2> | <h2>Parts overview</h2> | ||
− | <div><p> | + | <div><p>Through this project we have created a library of <i>Aspergillus</i> promoters and other fungal parts, which will make it easier for future iGEM-teams to work with filamentous fungi. These, along with other parts we have created or characterized, are described below.</p></div> |
</div> | </div> | ||
</div><!-- /end col-md-4 --> | </div><!-- /end col-md-4 --> | ||
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<div class="row flex-center sm-no-flex interlabspace"> | <div class="row flex-center sm-no-flex interlabspace"> | ||
<div class="column full_size"> | <div class="column full_size"> | ||
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<div class="interlabspace"> | <div class="interlabspace"> | ||
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p>This is a strong promoter for <i>Aspergillus niger</i> that has high activity in the exponential phase.</p> | + | <p>This is a strong promoter for <i>Aspergillus niger</i> that has high activity in the exponential phase. This is a synthetic exponential phase promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the glaA promoter from different <i>Aspergillus</i> spp., and the gene was chosen based on RNA-seq data from <i>Aspergillus niger</i>. The consensus promoter was expected to have really high expression. This version has “noise” added to the sequence to support a hypothesis that it was possible to turn down expression of the promoter.</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p>This is a medium strength constitutive promoter for <i>Aspergillus niger</i>.</p> | + | <p>This is a medium strength constitutive promoter for <i>Aspergillus niger</i>. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the sonB promoter from different <i>Aspergillus</i> spp., and the gene was chosen based on RNA-seq data from <i>Aspergillus niger</i>. [1] This version is a consensus sequence of the sonB promoters in Aspergillus and is expected to have a medium strength expression.</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p>This is a strong constitutive promoter for <i>Aspergillus niger</i> that has especially high activity in the lag phase. | + | <p>This is a strong constitutive promoter for <i>Aspergillus niger</i> that has especially high activity in the lag phase. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the gpdA promoter from different <i>Aspergillus</i> spp., and the gene was chosen based on RNA-seq data from <i>Aspergillus niger</i>. This version is a consensus sequence of the promoters in Aspergillus and is expected to have a high expression. |
</p> | </p> | ||
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p>This is a medium strength constitutive promoter for <i>Aspergillus niger</i>.</p> | + | <p>This is a medium strength constitutive promoter for <i>Aspergillus niger</i>. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the gpdA promoter from different <i>Aspergillus</i> spp., and the gene was chosen based on RNA-seq data from <i>Aspergillus niger</i>. The consensus promoter was expected to have high expression. This version has “noise” added to the sequence to support a hypothesis that it was possible to turn down expression of the promoter.</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p>This is a very strong constitutive promoter for <i>Aspergillus niger</i> that has especially high activity in the exponential growth phase.</p> | + | <p>This is a very strong constitutive promoter for <i>Aspergillus niger</i> that has especially high activity in the exponential growth phase. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the mstA promoter from different <i>Aspergillus</i> spp., and the gene was chosen based on RNA-seq data from <i>Aspergillus niger</i>. This version is a consensus sequence of the mstA promoters in Aspergillus and is expected to have a high constitutive expression.</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p>This is a weak constitutive promoter for <i>Aspergillus niger</i>.</p> | + | <p>This is a weak constitutive promoter for <i>Aspergillus niger</i>. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on an unknown promoter from different <i>Aspergillus</i> spp., and the gene was chosen based on RNA-seq data from <i>Aspergillus niger</i>. This version is a consensus sequence of the An07g08210 promoters in Aspergillus and is expected to have a weak constitutive expression.</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p>This is a strong promoter for <i>Aspergillus niger</i> that has high activity in the late exponential phase and stationary phase.</p> | + | <p>This is a strong promoter for <i>Aspergillus niger</i> that has high activity in the late exponential phase and stationary phase. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the gfaA promoter from different <i>Aspergillus</i> spp., and the gene was chosen based on RNA-seq data from <i>Aspergillus</i> niger. This version is a consensus sequence of the promoters in Aspergillus and is expected to have a high expression.</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p>This is a strong promoter for <i>Aspergillus niger</i> that has high activity in the stationary growth phase.</p> | + | <p>This is a strong promoter for <i>Aspergillus niger</i> that has high activity in the stationary growth phase. This is a synthetic stationary phase promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the hfbD promoter from different <i>Aspergillus</i> spp., and the gene was chosen based on RNA-seq data from <i>Aspergillus niger</i>. This is a consensus promoter and are expected to have high expression in the stationary phase.</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>This is a device for testing promoters by expressing mCherry.</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>This is a promoter for <i>Aspergillus niger</i> of unknown strength that is expected to have high expression in the exponential growth phase. This is a synthetic exponential phase promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the glaA promoter from different <i>Aspergillus</i> spp., and the gene was chosen based on RNA-seq data from <i>Aspergillus niger</i>. The consensus promoter was expected to have really high expression. This version has “noise” added to the sequence to support a hypothesis that it was possible to turn down expression of the promoter.</p> |
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</div> | </div> | ||
</div> | </div> | ||
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<!--############################################################################################# | <!--############################################################################################# | ||
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>This is a promoter for <i>Aspergillus niger</i> of unknown strength that is expected to have constitutive expression. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the sonB promoter from different <i>Aspergillus</i> spp., and the gene was chosen based on RNA-seq data from <i>Aspergillus niger</i>. This version is a consensus sequence of the sonB promoters in Aspergillus and is expected to have a medium strength expression</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>This is a very strong constitutive promoter for <i>Aspergillus niger</i> that has especially high activity in the exponential growth phase. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the gfaA promoter from different <i>Aspergillus</i> spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. This version has introduced stokastic noise compared to the consensus sequence of the promoters in Aspergillus and is therefore expected to have a lower expression.</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>This is a promoter for Aspergillus niger of unknown strength that should be active in the stationary phase. |
+ | This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the sonB promoter from different <i>Aspergillus</i> spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. [1] This version is a consensus sequence of an unknown gene's promoters in Aspergillus.</p> | ||
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>This is a promoter on unknown for Aspergillus niger that is expected to have high activity in the late exponential phase and stationary phase. |
+ | This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the gfaA promoter from different <i>Aspergillus</i> spp., and the gene was chosen based on RNA-seq data from <i>Aspergillus niger</i>. This version has introduced stokastic noise compared to the consensus sequence of the promoters in Aspergillus and is therefore expected to have a lower expression.</p> | ||
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<div class="partcombat"><p style="text-align:center;">Assembly Compatibility: </p><img class="assemblycombatimage" src="https://static.igem.org/mediawiki/2019/5/54/T--DTU-Denmark--gfaA_2comp.jpg" alt="The figure shows the compatibility of PLEAPgfaA_2" style="max-width: 170px;"> </div></div> | <div class="partcombat"><p style="text-align:center;">Assembly Compatibility: </p><img class="assemblycombatimage" src="https://static.igem.org/mediawiki/2019/5/54/T--DTU-Denmark--gfaA_2comp.jpg" alt="The figure shows the compatibility of PLEAPgfaA_2" style="max-width: 170px;"> </div></div> | ||
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</div> | </div> | ||
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>This part is the bacterial promoter <a href="http://parts.igem.org/Part:BBa_R0010" target="_blank">BBa_R0010</a> with MoClo level 0 compatible overhangs.</p> |
− | <p style="text-align:center;"> <img src="https://static.igem.org/mediawiki/2019/5/59/T--DTU-Denmark--moclo.jpg" alt="The figure shows the 238 basepair long MoClo promoter | + | <p style="text-align:center;"> <img src="https://static.igem.org/mediawiki/2019/5/59/T--DTU-Denmark--moclo.jpg" alt="The figure shows the 238 basepair long MoClo promoter stand-in" style="max-width: 721px;"> |
</p> | </p> | ||
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<div class="buttonshadow"> | <div class="buttonshadow"> | ||
− | <a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K3046018"><button class="collapsiblesmoll" style="border-right: 1px solid white;" title="Click here to watch part in registry">BBa_K3046018</button></a><button class="collapsiblebig">Ttef (Domesticated from <i> | + | <a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K3046018"><button class="collapsiblesmoll" style="border-right: 1px solid white;" title="Click here to watch part in registry">BBa_K3046018</button></a><button class="collapsiblebig">Ttef (Domesticated from <i>A. nidulans</i>)</button> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>Transcriptional terminator from <i>Aspergillus nidulans</i>. Domesticated to be in compliance with RFC[1000]</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>Protein secretion tag from the GlaA enzyme from <i>Aspergillus niger</i>.</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>This part is a superfolded, monomeric, stable green fluorescent protein.</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>This plasmid is designed as a shuttle vector to test different fungal promoters</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p>This is a plasmid containing an E.coli | + | <p>This is a plasmid containing an <i>E.coli</i> ORI, an ampicillin selection marker, and the AMA1 sequence. Inserts can be integrated into the plasmid by using a PacI/Nt.BbvCI USER assembly casette.</p> |
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<div class="buttonshadow"> | <div class="buttonshadow"> | ||
− | <a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K3046028"><button class="collapsiblesmoll" style="border-right: 1px solid white;" title="Click here to watch part in registry">BBa_K3046028</button></a><button class="collapsiblebig">PyrG CDS from <i> | + | <a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K3046028"><button class="collapsiblesmoll" style="border-right: 1px solid white;" title="Click here to watch part in registry">BBa_K3046028</button></a><button class="collapsiblebig">PyrG CDS from <i>A. fumigatus</i> </button> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>CThis coding sequence from <i>Aspergillus fumigatus</i> is encoding Orotidine 5'-phosphate decarboxylase.</p> |
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<div class="buttonshadow"> | <div class="buttonshadow"> | ||
− | <a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K3046029"><button class="collapsiblesmoll" style="border-right: 1px solid white;" title="Click here to watch part in registry">BBa_K3046029</button></a><button class="collapsiblebig">PyrG promoter from <i> | + | <a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K3046029"><button class="collapsiblesmoll" style="border-right: 1px solid white;" title="Click here to watch part in registry">BBa_K3046029</button></a><button class="collapsiblebig">PyrG promoter from <i>A. fumigatus</i> </button> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>Promoter from the PyrG selection marker gene from <i>Aspergillus fumigatus</i></p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>This part is designed to be inserted into pSB1C3 by Gibson cloning to make an integration plasmid for <i>Aspergillus niger</i>. The part is meant to be linearised by PCR prior to protoplast-mediated transformation, where the IS2 sites will lead to genomic integration of the insert (in this case just PyrG part BBa_K3046022) into the albA conidial pigment gene. Correct gene integration will, therefore, yield white/yellow conidia instead of the black conidia seen for the wildtype <i>A. niger</i>.</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>This part is designed to be inserted into pSB1C3 by Gibson cloning to make an integration plasmid for <i>Aspergillus niger</i>. The part is meant to be linearised by PCR prior to protoplast-mediated transformation, where the IS2 sites will lead to genomic integration of the insert (in this case just PyrG part BBa_K3046022) into the albA conidial pigment gene. Correct gene integration will therefore yield white/yellow conidia instead of the black conidia seen for the wildtype <i>A. niger</i>.</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>This part is a long sequence of DNA that are homologous to a part of the albA conidial pigment gene. This homology can be used for homologous recombination of genomic constructs flanked by this IS2 sequence (along with the downstream sequence BBa_K3046033).</p> |
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<div class="sizemargins"> | <div class="sizemargins"> | ||
− | <p> | + | <p>This part is a long sequence of DNA that are homologous to a part of the albA conidial pigment gene. This homology can be used for homologous recombination of genomic constructs flanked by this IS2 sequence (along with the upstream sequence BBa_K3046032).</p> |
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</div> | </div> | ||
Latest revision as of 02:21, 22 October 2019
Parts overview
Through this project we have created a library of Aspergillus promoters and other fungal parts, which will make it easier for future iGEM-teams to work with filamentous fungi. These, along with other parts we have created or characterized, are described below.