Difference between revisions of "Team:ICT-Mumbai/Composite Part"

 
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<h1>Parts</h1>
 
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<p>The aim of our project is to isolate the promoters of the genes that are upregulated in presence of root exudates of the crops and use these promoters in different applications. As a case study, we worked on construction of a positive feedback circuit to amplify the production of PhoD (alkaline phosphatase) in response to root exudates, during which we prepared several composite parts and intermediates and submitted them to registry.</p>
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<p>This year, Team ICT-Mumbai contributed four composite parts towards the iGEM Parts Registry. Our best composite part is described below:</p>
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<h4>Best Composite Part</h4>
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<h5>BBa_K2802001:</h5>
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<p>This part is a composite part composed of BBa_K274380, BBa_I746351 and BBa_K518012. A promoter can be cloned upstream of this part, and the PSP3 <I>pag</I> phage activator that will be produced will bind to the P<sub>F</sub> phage promoter to drive expression of the PSP3 <I>pag</I> phage activator, forming a genetic amplification circuit. RFP, which will be produced from the gene cloned downstream of the second <I>pag</I> gene, will act as a detectable output of this genetic amplification circuit.</p>
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<h5>Design</h5>
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BBa_K2802000 and BBa_K518012 were joined together using 3A Assembly.</p>
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<img style="display: block; margin: auto; height: 400px;" src="https://static.igem.org/mediawiki/2018/1/14/T--ICT-Mumbai--Best-composite.jpg">
 
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The 718 bp BBa_K2802000 (lane 2) and 828 bp BBa_K518012 parts were joined together to make 1554 bp BBa_K2802001 (lane 3). The plasmid backbone is pSB1C3. Plasmids were digested with EcoRI and PstI to release the parts and determine correct assembly of the parts.
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<h4>Other Composite Parts</h4>
 
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  <li>BBa_2802000:</li>
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<li><h5>BBa_K2802000:</h5></li>
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<p>This part is a composite part composed of BBa_K274380 and BBa_I746351. A promoter can be cloned upstream of this part, and the PSP3 <I>pag</I> phage activator that will be produced will bind to the PF phage promoter to drive expression of the PSP3 <I>pag</I> phage activator, forming a genetic amplifier. </p>
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<p>This part is a composite part composed of BBa_K274380 and BBa_I746351. A promoter can be cloned upstream of this part, and the PSP3 <I>pag</I> phage activator that will be produced will bind to the P<sub>F</sub> phage promoter to drive expression of the PSP3 <I>pag</I> phage activator, forming a genetic amplification circuit. </p>
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  <li>BBa_2802001:</li>
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<li><h5>BBa_K2802002:</h5></li>
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<p>This part is a composite part composed of BBa_K274380, BBa_I746351 and BBa_K518012. A promoter can be cloned upstream of this part, and the PSP3 <I>pag</I> phage activator that will be produced will bind to the PFF phage promoter to drive expression of the PSP3 <I>pag</I> phage activator, forming a genetic amplifier. <I>rfp</I>, which will be produced from the gene cloned downstream of the second <I>pag</I> gene, will act as a detectable output of this genetic amplifier.
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<p>This part is a composite part composed of BBa_K274371 and BBa_I746350. A promoter can be cloned upstream of this part, and the P2 <I>ogr</I> phage activator that will be produced will bind to the P<sub>O</sub> phage promoter to drive expression of the P2 <I>ogr</I> phage activator, forming a genetic amplification circuit.</p>
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  <li>BBa_2802002:</li>
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<li><h5>BBa_K2802003:</h5></li>
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<p>This part is a composite part composed of BBa_K274371 and BBa_I746350. A promoter can be cloned upstream of this part, and the P2 <I>ogr</I> phage activator that will be produced will bind to the PO phage promoter to drive expression of the P2 <I>ogr</I> phage activator, forming a genetic amplifier.</p>
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<p>This part is a composite part composed of BBa_K274371, BBa_I746350 and BBa_K518012. A promoter can be cloned upstream of this part, and the P2 <I>ogr</I> phage activator that will be produced will bind to the P<sub>O</sub> phage promoter to drive expression of the P2 <I>ogr</I> phage activator, forming a genetic amplification circuit. RFP, which will be produced from the gene cloned downstream of the second <I>ogr</I> gene, will act as a detectable output of this genetic amplification circuit.</p>
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<li>BBa_2802003:</li>
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<p>This part is a composite part composed of BBa_K274371, BBa_I746350 and BBa_K518012. A promoter can be cloned upstream of this part, and the P2 <I>ogr</I> phage activator that will be produced will bind to the PO phage promoter to drive expression of the P2 <I>ogr</I> phage activator, forming a genetic amplifier. RFP, which will be produced from the gene cloned downstream of the second <I>ogr</I> gene, will act as a detectable output of this genetic amplifier.</p>
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<h4>Composite Parts Table</h4>
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<center><groupparts>iGEM18 ICT-Mumbai</groupparts></center>
 
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Following are the intermediate parts submitted by the team to the iGEM registry</p>
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Latest revision as of 17:44, 17 October 2018

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Parts

This year, Team ICT-Mumbai contributed four composite parts towards the iGEM Parts Registry. Our best composite part is described below:

Best Composite Part

BBa_K2802001:

This part is a composite part composed of BBa_K274380, BBa_I746351 and BBa_K518012. A promoter can be cloned upstream of this part, and the PSP3 pag phage activator that will be produced will bind to the PF phage promoter to drive expression of the PSP3 pag phage activator, forming a genetic amplification circuit. RFP, which will be produced from the gene cloned downstream of the second pag gene, will act as a detectable output of this genetic amplification circuit.

Design

BBa_K2802000 and BBa_K518012 were joined together using 3A Assembly.


The 718 bp BBa_K2802000 (lane 2) and 828 bp BBa_K518012 parts were joined together to make 1554 bp BBa_K2802001 (lane 3). The plasmid backbone is pSB1C3. Plasmids were digested with EcoRI and PstI to release the parts and determine correct assembly of the parts.

Other Composite Parts

  1. BBa_K2802000:

    2. This part is a composite part composed of BBa_K274380 and BBa_I746351. A promoter can be cloned upstream of this part, and the PSP3 pag phage activator that will be produced will bind to the PF phage promoter to drive expression of the PSP3 pag phage activator, forming a genetic amplification circuit.

  2. BBa_K2802002:

    3. This part is a composite part composed of BBa_K274371 and BBa_I746350. A promoter can be cloned upstream of this part, and the P2 ogr phage activator that will be produced will bind to the PO phage promoter to drive expression of the P2 ogr phage activator, forming a genetic amplification circuit.

  3. BBa_K2802003:

    4. This part is a composite part composed of BBa_K274371, BBa_I746350 and BBa_K518012. A promoter can be cloned upstream of this part, and the P2 ogr phage activator that will be produced will bind to the PO phage promoter to drive expression of the P2 ogr phage activator, forming a genetic amplification circuit. RFP, which will be produced from the gene cloned downstream of the second ogr gene, will act as a detectable output of this genetic amplification circuit.

Composite Parts Table

<groupparts>iGEM18 ICT-Mumbai</groupparts>