Team:SDU-Denmark/Tour54

From 2013.igem.org

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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088000">BBa_K1088000 dxs (B. subtilis)</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088000">BBa_K1088000</a></span><br>
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The BioBrick contains the coding region of the dxs gene derived from the Gram-positive bacteria Bacillus subtilis. Dxs is the first enzyme in the MEP pathway converting pyruvate and <span class="tooltipLink">GAP</span> <span class="tooltip"><span class="tooltipHeader">GAP</span>Glyceraldehyde-3-phosphate</span> into <span class="tooltipLink">DXP.</span> <span class="tooltip"><span class="tooltipHeader">DXP</span>1-Deoxy-D-xylulose 5-phosphate</span> Has been sequenced.  
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The BioBrick contains the coding region of <span class="intro">the <span class="specialWord">dxs</span> gene</span> derived from the Gram-positive bacteria <span class="specialWord">Bacillus subtilis</span>. Dxs is the first enzyme in the MEP pathway converting pyruvate and <span class="tooltipLink">GAP</span> <span class="tooltip"><span class="tooltipHeader">GAP</span>Glyceraldehyde-3-phosphate</span> into <span class="tooltipLink">DXP.</span> <span class="tooltip"><span class="tooltipHeader">DXP</span>1-Deoxy-D-xylulose 5-phosphate</span> Has been sequenced.  
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088003">BBa_K1088003 HRT2</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088003">BBa_K1088003</a></span><br>
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The BioBrick contains the coding region of the HRT2 gene derived from Hevea brasiliensis and codon-optimized for Escherichia coli. HRT2 is the prenyl transferase that polymerizes <span class="tooltipLink">IPP</span> <span class="tooltip"><span class="tooltipHeader">IPP</span>Isopentenyl pyrophosphate</span> and <span class="tooltipLink">DMAPP</span> <span class="tooltip"><span class="tooltipHeader">DMAPP</span>Dimethylallyl pyrophosphate</span> into rubber. Has been sequenced and HRT2 function characterized by H<sup>1</sup>-NMR.
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The BioBrick contains the coding region of <span class="intro">the <span class="specialWord">HRT2</span> gene</span> derived from <span class="specialWord">Hevea brasiliensis</span> and codon-optimized for <span class="specialWord">Escherichia coli</span>. HRT2 is the prenyl transferase that polymerizes <span class="tooltipLink">IPP</span> <span class="tooltip"><span class="tooltipHeader">IPP</span>Isopentenyl pyrophosphate</span> and <span class="tooltipLink">DMAPP</span> <span class="tooltip"><span class="tooltipHeader">DMAPP</span>Dimethylallyl pyrophosphate</span> into rubber. Has been sequenced and HRT2 function characterized by H<sup>1</sup>-NMR.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088004">BBa_K1088004 ispG</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088004">BBa_K1088004</a></span><br>
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The BioBrick contains the coding region of the ispG gene derived from the Gram-negative bacteria Escherichia coli. ispG is the sixth enzyme in the MEP pathway converting <span class="tooltipLink">MEcPP</span> <span class="tooltip"><span class="tooltipHeader">MEcPP</span>2-C-methyl-D-erythritol 2,4-cyclopyrophosphate</span> into <span class="tooltipLink">HMB-PP.</span> <span class="tooltip"><span class="tooltipHeader">HMB-PP</span>(E)-4-Hydroxy-3-methyl-but-2-enyl pyrophosphate</span> Has been sequenced.
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The BioBrick contains the coding region of <span class="intro">the <span class="specialWord">ispG</span> gene</span> derived from the Gram-negative bacteria <span class="specialWord">Escherichia coli</span>. ispG is the sixth enzyme in the MEP pathway converting <span class="tooltipLink">MEcPP</span> <span class="tooltip"><span class="tooltipHeader">MEcPP</span>2-C-methyl-D-erythritol 2,4-cyclopyrophosphate</span> into <span class="tooltipLink">HMB-PP.</span> <span class="tooltip"><span class="tooltipHeader">HMB-PP</span>(E)-4-Hydroxy-3-methyl-but-2-enyl pyrophosphate</span> Has been sequenced.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088005">BBa_K1088005 araC</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088005">BBa_K1088005</a></span><br>
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The BioBrick contains the coding region of the araC gene derived from the Gram-negative bacteria Escherichia coli. AraC is a DNA-binding protein that regulates the transcription of operons involved in arabinose metabolism. With glucose present AraC functions as a repressor, and without glucose and with arabinose present it functions as an activator.
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The BioBrick contains the coding region of <span class="intro">the <span class="specialWord">araC</span> gene</span> derived from the Gram-negative bacteria <span class="specialWord">Escherichia coli</span>. AraC is a DNA-binding protein that regulates the transcription of operons involved in arabinose metabolism. With glucose present AraC functions as a repressor, and without glucose and with arabinose present it functions as an activator.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088018">BBa_K1088018 lacI</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088018">BBa_K1088018</a></span><br>
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The BioBrick contains the coding region of the lacI gene derived from the Gram-negative bacteria Escherichia coli. LacI is a DNA-binding protein that inhibits the transcription from the lac promoter when allolactose or <span class="tooltipLink">IPTG</span> <span class="tooltip"><span class="tooltipHeader">IPTG</span>Isopropyl β-D-1-thiogalactopyranoside</span> is absent. Has been sequenced and LacI function characterized by FACS and growth experiment.
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The BioBrick contains the coding region of <span class="intro">the <span class="specialWord">lacI</span> gene</span> derived from the Gram-negative bacteria <span class="specialWord">Escherichia coli</span>. LacI is a DNA-binding protein that inhibits the transcription from the lac promoter when allolactose or <span class="tooltipLink">IPTG</span> <span class="tooltip"><span class="tooltipHeader">IPTG</span>Isopropyl β-D-1-thiogalactopyranoside</span> is absent. Has been sequenced and LacI function characterized by <span class="tooltipLink">FACS.</span> <span class="tooltip"><span class="tooltipHeader">FACS</span>Fluorescence Activated Cell Sorting</span> and growth experiment.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088017">BBa_K1088017 Pcon-araC-term</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088017">BBa_K1088017</a></span><br>
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araC is being expressed from a constitutively active promoter. A terminator is put behind the coding region to prevent transcription of genes downstream of the activator. The device was used to check if we could enhance the control of the arabinose promoter. Has been sequenced and AraC function characterized by northern blot.
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<span class="intro">Pcon-araC-term:</span> <span class="specialWord">araC</span> is being expressed from a constitutively active promoter. A terminator is put behind the coding region to prevent transcription of genes downstream of the activator. The device was used to check if we could enhance the control of the arabinose promoter. Has been sequenced and AraC function characterized by northern blot.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088019">BBa_K1088019 Pcon-lacI(N)-term</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088019">BBa_K1088019</a></span><br>
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lacI is being expressed from a constitutively active promoter. A terminator is put behind the coding region to prevent transcription of genes downstream of the repressor. The device was used to enable us to control the lactose promoter. This device proved be most effective together for expression control. LacI(N) function characterized by GFP fusion using FACS and growth experiment.
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<span class="intro">Pcon-lacI(N)-term:</span> <span class="specialWord">lacI</span> is being expressed from a constitutively active promoter. A terminator is put behind the coding region to prevent transcription of genes downstream of the repressor. The device was used to enable us to control the lactose promoter. This device proved be most effective together for expression control. LacI(N) function characterized by GFP fusion using <span class="tooltipLink">FACS.</span> <span class="tooltip"><span class="tooltipHeader">FACS</span>Fluorescence Activated Cell Sorting</span> and growth experiment.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088020">BBa_K1088020 Pcon-lacI:LVA-term</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088020">BBa_K1088020</a></span><br>
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lacI:LVA (BBa_C0012) is being expressed from a constitutively active promoter. A terminator is put behind the coding region to prevent transcription of genes downstream of the repressor. The LVA-tag is a tag for degradation, and thus there is increased turnover of the protein. The device is meant to enable us to control the lactose promoter. However natural LacI proved to be more effective than the LVA-tagged. Has been sequenced and LacI:LVA function characterized by GFP fusion using FACS and growth experiment.
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<span class="intro">Pcon-lacI:LVA-term:</span> <span class="specialWord">lacI:LVA</span> (<a class="dialogLink" href="http://parts.igem.org/Part:Ba_C0012">BBa_C0012</a>) is being expressed from a constitutively active promoter. A terminator is put behind the coding region to prevent transcription of genes downstream of the repressor. The LVA-tag is a tag for degradation, and thus there is increased turnover of the protein. The device is meant to enable us to control the lactose promoter. However natural LacI proved to be more effective than the LVA-tagged. Has been sequenced and LacI:LVA function characterized by GFP fusion using FACS and growth experiment.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088006">BBa_K1088006 Pcon-dxs (B. subtilis)-amilCP</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088006">BBa_K1088006</a></span><br>
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This device expresses the dxs gene derived from B. subtilis linked to GFP, and is under the control of the lactose promoter. AmilCP proved to be a poor fusion protein for the Dxs protein. Has been sequenced.
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<span class="intro">Pcon-<span class="specialWord">dxs (B. subtilis)</span>-amilCP</span> expresses the <span class="specialWord">dxs</span> gene derived from   <span class="specialWord">Bacillus subtilis</span> linked to GFP, and is under the control of the lactose promoter. AmilCP proved to be a poor fusion protein for the Dxs protein. Has been sequenced.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088007">BBa_K1088007 Plac-dxs (E. coli)-GFP</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088007">BBa_K1088007</a></span><br>
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This device expresses the dxs gene derived from  E. coli linked to GFP, and is under the control of the lactose promoter. The device was used to check the expression level of the E. coli dxs gene under various conditions. Has been sequenced.
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<span class="intro">Plac-<span class="specialWord">dxs (E. coli)</span>-GFP</span> expresses the <span class="specialWord">dxs</span> gene derived from  <span class="specialWord">Escherichia coli</span> linked to GFP, and is under the control of the lactose promoter. The device was used to check the expression level of the <span class="specialWord">E. coli dxs</span> gene under various conditions. Has been sequenced.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088008">BBa_K1088008 Plac-dxs (B. subtilis):GFP</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088008">BBa_K1088008</a></span><br>
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This device expresses the dxs gene derived from B. subtilis linked to GFP, and is under the control of the lactose promoter. The device is was used to check the expression level of the B.subtilis dxs gene under various conditions. Has been sequenced and Plac function characterized by GFP fusion using FACS and growth experiment.
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<span class="intro">Plac-<span class="specialWord">dxs (B. subtilis)</span>:GFP</span> expresses the <span class="specialWord">dxs</span> gene derived from <span class="specialWord">Bacillus subtilis</span> linked to GFP, and is under the control of the lactose promoter. The device is was used to check the expression level of the <span class="specialWord">B.subtilis dxs</span> gene under various conditions. Has been sequenced and Plac function characterized by GFP fusion using <span class="tooltipLink">FACS.</span> <span class="tooltip"><span class="tooltipHeader">FACS</span>Fluorescence Activated Cell Sorting</span> and growth experiment.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088009">BBa_K1088009 Pcon-lacI:LVA-Plac-dxs (B. subtilis)-GFP</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088009">BBa_K1088009</a></span><br>
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This device expresses the dxs gene derived from  B. subtilis linked to GFP, and is under the control of the lactose promoter. The device to check the expression level of the B.subtilis dxs gene under various conditions. Our LacI:LVA (BBa_K1088020) device (with a constitutive promoter was added to optimize the expression control through the lactose promoter. Natural LacI proved to be more efficient, though. Has been sequenced. LacI and Plac function characterized by GFP fusion using FACS and growth experiment.  
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<span class="intro">Pcon-<span class="specialWord">lacI:LVA</span>-Plac-<span class="specialWord">dxs (B. subtilis)</span>-GFP</span> expresses the <span class="specialWord">dxs</span> gene derived from  <span class="specialWord">B. subtilis</span> linked to GFP, and is under the control of the lactose promoter. The device to check the expression level of the <span class="specialWord">Bacillus subtilis</span> dxs gene under various conditions. Our LacI:LVA (<a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088020">BBa_K1088020</a>) device (with a constitutive promoter was added to optimize the expression control through the lactose promoter. Natural LacI proved to be more efficient, though. Has been sequenced. LacI and Plac function characterized by GFP fusion using FACS and growth experiment.  
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088010">BBa_K1088010 Pcon-lacI:LVA-term-Plac-dxs (E. coli)-GFP</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088010">BBa_K1088010</a></span><br>
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This device expresses the dxs gene derived from  E. coli linked to GFP, and is under the control of the lactose promoter. The device is meant for us to check the expression level of the dxs gene under various conditions. Our LacI:LVA (BBa_K1088020) device (with a constitutive promoter was added to optimize the expression control through the lactose promoter.
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<span class="intro">Pcon-<span class="specialWord">lacI:LVA</span>-term-Plac-<span class="specialWord">dxs (E. coli)</span>-GFP</span> expresses the <span class="specialWord">dxs</span> gene derived from  <span class="specialWord">Escherichia coli</span> linked to GFP, and is under the control of the lactose promoter. The device is meant for us to check the expression level of the <span class="specialWord">dxs</span> gene under various conditions. Our LacI:LVA (<a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088020">BBa_K1088020</a>) device (with a constitutive promoter was added to optimize the expression control through the lactose promoter.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088026">BBa_K1088026 Pcon-lacI(N)-Plac-dxs (B. subtilis)-GFP</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088026">BBa_K1088026</a></span><br>
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This device expresses the dxs gene derived from B. subtilis linked to GFP, and is under the control of the lactose promoter. The device is meant for us to check the expression level of the dxs gene under various conditions. Furthermore the lacI gene with a constitutive promoter has been added to optimize the expression control through the lactose promoter. This device proved to have the most efficient expression control (see results for more detail). Has been sequenced. LacI and Plac function characterized by GFP fusion using FACS and growth experiment.  
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<span class="intro">Pcon-<span class="specialWord">lacI(N)</span>-Plac-<span class="specialWord">dxs (B. subtilis)</span>-GFP</span> expresses the <span class="specialWord">dxs</span> gene derived from   <span class="specialWord">Bacillus subtilis</span> linked to GFP, and is under the control of the lactose promoter. The device is meant for us to check the expression level of the dxs gene under various conditions. Furthermore the <span class="specialWord">lacI</span> gene with a constitutive promoter has been added to optimize the expression control through the lactose promoter. This device proved to have the most efficient expression control (see results for more detail). Has been sequenced. LacI and Plac function characterized by GFP fusion using FACS and growth experiment.  
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088011">BBa_K1088011 Plac-dxs(B. subtilis)</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088011">BBa_K1088011</a></span><br>
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This device expresses the dxs gene derived from  B. subtilis, and is under the control of the lactose promoter. The device is meant for us to increase the amount of IPP and DMAPP in the cell. Plac function characterized by GFP fusion using FACS and growth experiment.  
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<span class="intro">Plac-<span class="specialWord">dxs (B. subtilis)</span></span> expresses the <span class="specialWord">dxs</span> gene derived from  <span class="specialWord">B. subtilis</span>, and is under the control of the lactose promoter. The device is meant for us to increase the amount of IPP and DMAPP in the cell. Plac function characterized by GFP fusion using <span class="tooltipLink">FACS.</span> <span class="tooltip"><span class="tooltipHeader">FACS</span>Fluorescence Activated Cell Sorting</span> and growth experiment.  
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088012">BBa_K1088012 Plac-dxs(E. coli)</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088012">BBa_K1088012</a></span><br>
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This device expresses the dxs gene (BBa_K118000) derived from  E. coli, and is under the control of the lactose promoter. The device is meant for us to increase the amount of IPP and DMAPP in the cell. Has been sequenced.
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<span class="intro">Plac-<span class="specialWord">dxs (E. coli)</span></span> expresses the <span class="specialWord">dxs</span> gene (<a class="dialogLink" href="http://parts.igem.org/Part:BBa_K118000" title="">BBa_K118000</a>) derived from  <span class="specialWord">E. coli</span>, and is under the control of the lactose promoter. The device is meant for us to increase the amount of IPP and DMAPP in the cell. Has been sequenced.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088013">BBa_K1088013 Pcon-lacI:LVA-term-Plac-dxs(B. subtilis)</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088013">BBa_K1088013</a></span><br>
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This device expresses the dxs gene derived from  B. subtilis, and is under the control of the lactose promoter. The device is meant for us to increase the amount of <span class="tooltipLink">IPP</span> <span class="tooltip"><span class="tooltipHeader">IPP</span>Isopentenyl pyrophosphate</span> and <span class="tooltipLink">DMAPP</span> <span class="tooltip"><span class="tooltipHeader">DMAPP</span>Dimethylallyl pyrophosphate</span> in the cell. Furthermore the lacI:LVA gene with a constitutive promoter was added to optimize the expression control through the lactose promoter. Natural LacI proved to be more efficient, though. Has been sequenced.
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<span class="intro">Pcon-<span class="specialWord">lacI:LVA</span>-term-Plac-<span class="specialWord">dxs (B. subtilis)</span></span> expresses the dxs gene derived from  <span class="specialWord">B. subtilis</span>, and is under the control of the lactose promoter. The device is meant for us to increase the amount of <span class="tooltipLink">IPP</span> <span class="tooltip"><span class="tooltipHeader">IPP</span>Isopentenyl pyrophosphate</span> and <span class="tooltipLink">DMAPP</span> <span class="tooltip"><span class="tooltipHeader">DMAPP</span>Dimethylallyl pyrophosphate</span> in the cell. Furthermore the <span class="specialWord">lacI:LVA</span> gene with a constitutive promoter was added to optimize the expression control through the lactose promoter. Natural LacI proved to be more efficient, though. Has been sequenced.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088014">BBa_K1088014 Pcon-lacI:LVA-term-Plac-dxs(E. coli)</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088014">BBa_K1088014</a></span><br>
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This device expresses the dxs gene derived from  E. coli, and is under the control of the lactose promoter. The device is meant for us to increase the amount of IPP and DMAPP in the cell. Furthermore the lacI-LVA gene with a constitutive promoter has been added to optimize the expression control through the lactose promoter.
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<span class="intro">Pcon-<span class="specialWord">lacI:LVA</span>-term-Plac-<span class="specialWord">dxs (E. coli)</span></span> expresses the <span class="specialWord">dxs</span> gene derived from  <span class="specialWord">E. coli</span>, and is under the control of the lactose promoter. The device is meant for us to increase the amount of IPP and DMAPP in the cell. Furthermore the <span class="specialWord">lacI-LVA</span> gene with a constitutive promoter has been added to optimize the expression control through the lactose promoter.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088015">BBa_K1088015 Pcon-lacI:LVA-term-Plac-dxs(B. subtilis)-ispG</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088015">BBa_K1088015</a></span><br>
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This device expresses the dxs gene derived from  B. subtilis, and is under the control of the lactose promoter.The device was build to increase the amount of IPP and DMAPP in the cell if the first rate limiting step was overcome.  Furthermore the lacI:LVA gene with a constitutive promoter has been added to optimize the expression control through the lactose promoter (see results for description LacI-LVA efficiency).
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<span class="intro">Pcon-<span class="specialWord">lacI:LVA</span>-term-Plac-<span class="specialWord">dxs (B. subtilis)</span>-ispG</span> expresses the <span class="specialWord">dxs</span> gene derived from  <span class="specialWord">B. subtilis</span>, and is under the control of the lactose promoter. The device was build to increase the amount of IPP and DMAPP in the cell if the first rate limiting step was overcome.  Furthermore the <span class="specialWord">lacI:LVA</span> gene with a constitutive promoter has been added to optimize the expression control through the lactose promoter (see results for description LacI-LVA efficiency).
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088016">BBa_K1088016 Pcon-araC-term-Para-HRT2-(3xFLAG)</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088016">BBa_K1088016</a></span><br>
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This device expresses the HRT2 gene derived from Hevea brasiliensis, and is under the control of the arabinose promoter. The device was made to enable the bacteria to polymerize IPP and DMAPP into rubber. Furthermore the arabinose promoter regulator AraC has been added to check if it would enhance the expression control of arabinose promoter . It did not seem to improve expression control. Has been sequenced. AraC and Para function characterized by Northern blot and HRT2 function characterized by H<sup>1</sup>-NMR.
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<span class="intro">Pcon-<span class="specialWord">araC</span>-term-Para-<span class="specialWord">HRT2-(3xFLAG)</span></span> expresses the <span class="specialWord">HRT2</span> gene derived from <span class="specialWord">Hevea brasiliensis</span>, and is under the control of the arabinose promoter. The device was made to enable the bacteria to polymerize IPP and DMAPP into rubber. Furthermore the arabinose promoter regulator AraC has been added to check if it would enhance the expression control of arabinose promoter. It did not seem to improve expression control. Has been sequenced. AraC and Para function characterized by Northern blot and HRT2 function characterized by H<sup>1</sup>-NMR.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088024">BBa_K1088024 Para-HRT2-(3xFLAG)</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088024">BBa_K1088024</a></span><br>
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This device expresses the HRT2 gene derived from Hevea brasiliensis, and is under the control of the arabinose promoter. The device is meant to enable the bacteria to polymerize IPP and DMAPP into rubber. Has been sequenced. AraC and Para function characterized by Northern blot and HRT2 function characterized by H<sup>1</sup>-NMR.
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<span class="intro">Para-<span class="specialWord">HRT2-(3xFLAG)</span></span> expresses the <span class="specialWord">HRT2</span> gene derived from <span class="specialWord">Hevea brasiliensis</span>, and is under the control of the arabinose promoter. The device is meant to enable the bacteria to polymerize IPP and DMAPP into rubber. Has been sequenced. AraC and Para function characterized by Northern blot and HRT2 function characterized by H<sup>1</sup>-NMR.
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088027">BBa_K1088027 Pcon-lacI(N)-Plac-dxs (B. subtilis)</a></span><br>
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<span class="intro"><a class="dialogLink" href="http://parts.igem.org/Part:BBa_K1088027">BBa_K1088027</a></span><br>
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This device expresses the dxs gene derived from  B. subtilis, and is under the control of the lactose promoter. The device is meant for us to increase the amount of IPP and DMAPP in the cell. Furthermore the lacI gene with a constitutive promoter was added to optimize the expression control through the lactose promoter. This addition proved to have the most efficient expression control. LacI and Plac function characterized by GFP fusion using FACS and growth experiment.   
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<span class="intro">Pcon-<span class="specialWord">lacI(N)</span>-Plac-<span class="specialWord">dxs (B. subtilis)</span></span> expresses the <span class="specialWord">dxs</span> gene derived from  <span class="specialWord">B. subtilis</span>, and is under the control of the lactose promoter. The device is meant for us to increase the amount of IPP and DMAPP in the cell. Furthermore the <span class="specialWord">lacI</span> gene with a constitutive promoter was added to optimize the expression control through the lactose promoter. This addition proved to have the most efficient expression control. LacI and Plac function characterized by GFP fusion using <span class="tooltipLink">FACS.</span> <span class="tooltip"><span class="tooltipHeader">FACS</span>Fluorescence Activated Cell Sorting</span> and growth experiment.   
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Latest revision as of 23:54, 28 October 2013

Submitted Parts

And so, the iGEM parts registry grows

Browse through the tabs below to get a complete picture of our submitted parts. Feel free to follow the links to parts registry, where you can find information on sequencing, characterization, etc.


Coding
BBa_K1088000
The BioBrick contains the coding region of the dxs gene derived from the Gram-positive bacteria Bacillus subtilis. Dxs is the first enzyme in the MEP pathway converting pyruvate and GAP GAPGlyceraldehyde-3-phosphate into DXP. DXP1-Deoxy-D-xylulose 5-phosphate Has been sequenced.

BBa_K1088003
The BioBrick contains the coding region of the HRT2 gene derived from Hevea brasiliensis and codon-optimized for Escherichia coli. HRT2 is the prenyl transferase that polymerizes IPP IPPIsopentenyl pyrophosphate and DMAPP DMAPPDimethylallyl pyrophosphate into rubber. Has been sequenced and HRT2 function characterized by H1-NMR.

BBa_K1088004
The BioBrick contains the coding region of the ispG gene derived from the Gram-negative bacteria Escherichia coli. ispG is the sixth enzyme in the MEP pathway converting MEcPP MEcPP2-C-methyl-D-erythritol 2,4-cyclopyrophosphate into HMB-PP. HMB-PP(E)-4-Hydroxy-3-methyl-but-2-enyl pyrophosphate Has been sequenced.

BBa_K1088005
The BioBrick contains the coding region of the araC gene derived from the Gram-negative bacteria Escherichia coli. AraC is a DNA-binding protein that regulates the transcription of operons involved in arabinose metabolism. With glucose present AraC functions as a repressor, and without glucose and with arabinose present it functions as an activator.

BBa_K1088018
The BioBrick contains the coding region of the lacI gene derived from the Gram-negative bacteria Escherichia coli. LacI is a DNA-binding protein that inhibits the transcription from the lac promoter when allolactose or IPTG IPTGIsopropyl β-D-1-thiogalactopyranoside is absent. Has been sequenced and LacI function characterized by FACS. FACSFluorescence Activated Cell Sorting and growth experiment.
Regulatory devices
BBa_K1088017
Pcon-araC-term: araC is being expressed from a constitutively active promoter. A terminator is put behind the coding region to prevent transcription of genes downstream of the activator. The device was used to check if we could enhance the control of the arabinose promoter. Has been sequenced and AraC function characterized by northern blot.

BBa_K1088019
Pcon-lacI(N)-term: lacI is being expressed from a constitutively active promoter. A terminator is put behind the coding region to prevent transcription of genes downstream of the repressor. The device was used to enable us to control the lactose promoter. This device proved be most effective together for expression control. LacI(N) function characterized by GFP fusion using FACS. FACSFluorescence Activated Cell Sorting and growth experiment.

BBa_K1088020
Pcon-lacI:LVA-term: lacI:LVA (BBa_C0012) is being expressed from a constitutively active promoter. A terminator is put behind the coding region to prevent transcription of genes downstream of the repressor. The LVA-tag is a tag for degradation, and thus there is increased turnover of the protein. The device is meant to enable us to control the lactose promoter. However natural LacI proved to be more effective than the LVA-tagged. Has been sequenced and LacI:LVA function characterized by GFP fusion using FACS and growth experiment.
Reporter fusions
BBa_K1088006
Pcon-dxs (B. subtilis)-amilCP expresses the dxs gene derived from Bacillus subtilis linked to GFP, and is under the control of the lactose promoter. AmilCP proved to be a poor fusion protein for the Dxs protein. Has been sequenced.

BBa_K1088007
Plac-dxs (E. coli)-GFP expresses the dxs gene derived from Escherichia coli linked to GFP, and is under the control of the lactose promoter. The device was used to check the expression level of the E. coli dxs gene under various conditions. Has been sequenced.

BBa_K1088008
Plac-dxs (B. subtilis):GFP expresses the dxs gene derived from Bacillus subtilis linked to GFP, and is under the control of the lactose promoter. The device is was used to check the expression level of the B.subtilis dxs gene under various conditions. Has been sequenced and Plac function characterized by GFP fusion using FACS. FACSFluorescence Activated Cell Sorting and growth experiment.

BBa_K1088009
Pcon-lacI:LVA-Plac-dxs (B. subtilis)-GFP expresses the dxs gene derived from B. subtilis linked to GFP, and is under the control of the lactose promoter. The device to check the expression level of the Bacillus subtilis dxs gene under various conditions. Our LacI:LVA (BBa_K1088020) device (with a constitutive promoter was added to optimize the expression control through the lactose promoter. Natural LacI proved to be more efficient, though. Has been sequenced. LacI and Plac function characterized by GFP fusion using FACS and growth experiment.

BBa_K1088010
Pcon-lacI:LVA-term-Plac-dxs (E. coli)-GFP expresses the dxs gene derived from Escherichia coli linked to GFP, and is under the control of the lactose promoter. The device is meant for us to check the expression level of the dxs gene under various conditions. Our LacI:LVA (BBa_K1088020) device (with a constitutive promoter was added to optimize the expression control through the lactose promoter.

BBa_K1088026
Pcon-lacI(N)-Plac-dxs (B. subtilis)-GFP expresses the dxs gene derived from Bacillus subtilis linked to GFP, and is under the control of the lactose promoter. The device is meant for us to check the expression level of the dxs gene under various conditions. Furthermore the lacI gene with a constitutive promoter has been added to optimize the expression control through the lactose promoter. This device proved to have the most efficient expression control (see results for more detail). Has been sequenced. LacI and Plac function characterized by GFP fusion using FACS and growth experiment.
Constitutively active production devices
BBa_K1088011
Plac-dxs (B. subtilis) expresses the dxs gene derived from B. subtilis, and is under the control of the lactose promoter. The device is meant for us to increase the amount of IPP and DMAPP in the cell. Plac function characterized by GFP fusion using FACS. FACSFluorescence Activated Cell Sorting and growth experiment.

BBa_K1088012
Plac-dxs (E. coli) expresses the dxs gene (BBa_K118000) derived from E. coli, and is under the control of the lactose promoter. The device is meant for us to increase the amount of IPP and DMAPP in the cell. Has been sequenced.
Regulable production devices
BBa_K1088013
Pcon-lacI:LVA-term-Plac-dxs (B. subtilis) expresses the dxs gene derived from B. subtilis, and is under the control of the lactose promoter. The device is meant for us to increase the amount of IPP IPPIsopentenyl pyrophosphate and DMAPP DMAPPDimethylallyl pyrophosphate in the cell. Furthermore the lacI:LVA gene with a constitutive promoter was added to optimize the expression control through the lactose promoter. Natural LacI proved to be more efficient, though. Has been sequenced.

BBa_K1088014
Pcon-lacI:LVA-term-Plac-dxs (E. coli) expresses the dxs gene derived from E. coli, and is under the control of the lactose promoter. The device is meant for us to increase the amount of IPP and DMAPP in the cell. Furthermore the lacI-LVA gene with a constitutive promoter has been added to optimize the expression control through the lactose promoter.

BBa_K1088015
Pcon-lacI:LVA-term-Plac-dxs (B. subtilis)-ispG expresses the dxs gene derived from B. subtilis, and is under the control of the lactose promoter. The device was build to increase the amount of IPP and DMAPP in the cell if the first rate limiting step was overcome. Furthermore the lacI:LVA gene with a constitutive promoter has been added to optimize the expression control through the lactose promoter (see results for description LacI-LVA efficiency).

BBa_K1088016
Pcon-araC-term-Para-HRT2-(3xFLAG) expresses the HRT2 gene derived from Hevea brasiliensis, and is under the control of the arabinose promoter. The device was made to enable the bacteria to polymerize IPP and DMAPP into rubber. Furthermore the arabinose promoter regulator AraC has been added to check if it would enhance the expression control of arabinose promoter. It did not seem to improve expression control. Has been sequenced. AraC and Para function characterized by Northern blot and HRT2 function characterized by H1-NMR.

BBa_K1088024
Para-HRT2-(3xFLAG) expresses the HRT2 gene derived from Hevea brasiliensis, and is under the control of the arabinose promoter. The device is meant to enable the bacteria to polymerize IPP and DMAPP into rubber. Has been sequenced. AraC and Para function characterized by Northern blot and HRT2 function characterized by H1-NMR.

BBa_K1088027
Pcon-lacI(N)-Plac-dxs (B. subtilis) expresses the dxs gene derived from B. subtilis, and is under the control of the lactose promoter. The device is meant for us to increase the amount of IPP and DMAPP in the cell. Furthermore the lacI gene with a constitutive promoter was added to optimize the expression control through the lactose promoter. This addition proved to have the most efficient expression control. LacI and Plac function characterized by GFP fusion using FACS. FACSFluorescence Activated Cell Sorting and growth experiment.