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| − | This part is contained in the Green Expansion, a range of parts from <a href="https:// 2019.igem.org/ Team: Marburg"> iGEM Marburg 2019</a> that enables users of the Marburg Collection 2. 0 to design MoClo compatible vectors for cyanobacteria as well as to engineer the genome of several cyanobacterial species. | + | This BioBrick is part of the measurement entry vectors for the characterization of BioBricks in cyanobacteria (Part range <a href="https://parts .igem.org/Part:BBa_K3228073"> BBa_K3228073</a> to <a href="https:// parts.igem.org/ Part: BBa_K3228075"> BBa_K3228075</a> as well as <a href="https://parts.igem.org/Part:BBa_K3228090"> BBa_K3228090</a>) by the iGEM Team Marburg 2019. They contain our MoClo compatible shuttle vector for cyanobacteria <a href="https://parts.igem.org/Part:BBa_K3228069"> BBa_K3228069</a> and are therefore the only MoClo based vectors for the characterization of BioBricks in cyanobacteria. These pre assembled LVL 1 plasmids contain a place holder < a href="https:/ /parts.igem.org/Part:BBa_K3228060"> (compare BBa_K3228060)</ a> for their respective BioBrick type that acts as a Dropout to quickly and effortlessly insert any part of the same type for an easy characterization. |
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| − | ===The Green Expansion===
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| − | The Green Expansion is an addition of parts to the Marburg Collection 2.0 <a href="http://2018.igem.org/Team:Marburg/Design">(See: Design of the Marburg Collection)</a> that features the world's first MoClo compatible shuttle vector for cyanobacteria. <a href="https://parts.igem.org/Part:BBa_K3228069">BBa_K3228069</a>
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| − | | + | <div style="width:100%;display:flex;flex-direction:row;flex-wrap: wrap; justify-content:space-evenly; align-items:center;">[[File:GE LVL2 measurementvectors.png|500px|thumb|left|'''Figure 1''': Design of the standardized measurement vectors for terminator BioBrick characterization ]] |
| − | <div style="width:100%;display:flex;flex-direction:row;flex-wrap: wrap; justify-content:space-evenly; align-items:center;"> | + | |
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| − | [[File:GE LVL1 ori.png|500px|thumb|left|'''Figure 1''': Design of the first MoClo compatible shuttle vector for cyanobacteria for LVL 1 constructs. This can be used for the integration of simple genetic modules. ]] | + | |
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| − | [[File:GE LVL2 ori.png|500px|thumb|left|'''Figure 2''': Design of the first MoClo compatible shuttle vector for cyanobacteria for LVL 2 constructs. This can be used for the integration of complex genetic devices. ]]
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| − | The Green Expansion also offers all the parts needed for the genomic integration of one or multiple genes in cyanobacteria. This M.E.G.A. (Modularized Engineering of Genome Areas) kit convinces with a striking flexibility and a very intuitive workflow for the de novo assembly of your plasmid of choice. It encompasses five different neutral integration sites to choose from: three conventional sites frequently used in the cyanobacterial community (NSI to NSIII) as well as our own rationally designed artificial Neutral integration Site options a.N.S.o. 1 and 2 <a href="https://2019.igem.org/Team:Marburg/Design">(See: Finding new artificial Neutral integration Site options).</a>These sites show no transcriptional activity from neighboring regions according to RNA-seq data and are therefore completely orthogonal. Additionally we offer four different antibiotic markers to use (chloramphenicol, gentamicin, spectinomycin and kanamycin). With the Green Expansion up to 20 genes can be introduced into a cyanobacterial strain.
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| − | <div style="width:100%;display:flex;flex-direction:row;flex-wrap: wrap; justify-content:space-evenly; align-items:center;"> | + | In our <a href="https://2019.igem.org/Team:Marburg/Results">results</a>we show how these measurement entry vectors can save a lot of effort and money when characterizing a greater library of parts. Additionally the usage of the same entry vector for each measurement will aid in greater comparability and reproducibility. |
| | + | For greater comparability across other data sets we decided to use the same Biobricks as in measuring the toolbox for <a href="http://2018.igem.org/Team:Marburg/Results"> Vibrio natriegens in the last year</a>. |
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| − | [[File:GE genomic integration.png|500px|thumb|left|'''Figure 3''': Overview over the modularized editing of genome area kit (M.E.G.A. kit) ]]
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| − | Thanks to the flexible design this expansion can also be used for the genomic modification of any chassis after the introduction of a new species specific LVL 0 integration sites to our Marburg Collection 2.0. As the workflow to build new homologies is a bit more intricate compared to the one pot on step assembly of our other parts due to the internal BsmBI cutting site, we described the workflow for that in our design section <a href="https://2019.igem.org/Team:Marburg/Design">(See: Design of neutral integration sites).</a>
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| − | The Green Expansion proves a valuable addition to our Marburg Collection 2.0 and to the iGEM Registry of Standard Biological Parts. It services users of our chassis and other cyanobacterial strains with a useful tool for genomic modifications but it also contributes a shell that can be used to modify any other model organism as well.
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| − | These parts are compatible with the RCF [1000] standard and can be used in any part collection that uses the PhytoBrick standard of overhangs. For more information we recommend to head over to <a href="http://2018.igem.org/Team:Marburg/Design">Design of the Marburg Collection iGEM Marburg 2018</a>. | + | |
| | + | These parts are compatible with the RCF [1000] standard and can be used in any part collection that uses the PhytoBrick standard of overhangs. For more information we recommend to head over to <a href="http://2018.igem.org/Team:Marburg/Design">Design of the Marburg Collection</a>. |
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| − | ===Parts of the Green Expansion=== | + | ===Available standard measurement vectors=== |
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| | <div class="PCListIcon" style="display:flex;flex-direction:row; flex-wrap: nowrap; justify-content:space-evenly; align-items:flex-start;font-size:82%;"> | | <div class="PCListIcon" style="display:flex;flex-direction:row; flex-wrap: nowrap; justify-content:space-evenly; align-items:flex-start;font-size:82%;"> |
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| − | [[File:Icon hConnector5.png|90px|thumb|none| | + | [[File:Icon MesurementVector.png|90px|thumb|none| |
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228000">K3228000 </a> (aNSo1 integration up) </li> | + | <li> <a href="https://parts.igem.org/Part:BBa_K3228073">K3228073 </a> <br>(promoter entry vector ) </li> |
| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228001">K3228001 </a> (aNSo2 integration up) </li>
| + | <li> <a href="https://parts.igem.org/Part:BBa_K3228074">K3228074 </a> <br>(rbs entry vector) </li> |
| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228020">K3228020 </a> <br> (NS1 up) </li> | + | <li> <a href="https://parts.igem.org/Part:BBa_K3228075">K3228075 </a> <br>(CDS entry vector) </li> |
| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228021">K3228021 </a> <br> (NS2 up) </li> | + | <li> <a href="https://parts.igem.org/Part:BBa_K3228090">K3228090 </a> <br>(terminator entry vector) </li> |
| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228022">K3228022 </a> <br> (NS3 up) </li> | + | |
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228004">K3228004 </a> (shortTB0010) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228005">K3228005 </a> (shortTB0015) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228006">K3228006 </a> (shortTB1002) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228007">K3228007 </a> (shortTB1003) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228008">K3228008 </a> (shortTB1004) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228009">K3228009 </a> (shortTB1005) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228010">K3228010 </a> (shortTB1006) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228011">K3228011 </a> (shortTB1007) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228012">K3228012 </a> (shortTB1008) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228013">K3228013 </a> (shortTB1009) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228014">K3228014 </a> (shortTB1010) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228015">K3228015 </a> (shortDummy) </li>
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| − | <li><a href="https://parts.igem.org/Part:BBa_K3228016">K3228016 </a> (SpecRes_short) </li>
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| − | <li><a href="https://parts.igem.org/Part:BBa_K3228017">K3228017 </a> <br> (CmlRes_short) </li>
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| − | <li><a href="https://parts.igem.org/Part:BBa_K3228018">K3228018 </a> <br> (TetRes_short) </li>
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| − | <li><a href="https://parts.igem.org/Part:BBa_K3228019">K3228019 </a> <br> (KanRes_short) </li>
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| − | <li><a href="https://parts.igem.org/Part:BBa_K3228027">K3228027 </a> <br> (GenRes_short) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228002">K3228002 </a> (aNSo1 integration down) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228003">K3228003 </a> (aNSo1 integration down) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228023">K3228023 </a> <br> (NS1 down) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228024">K3228024 </a> <br> (NS2 down) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228025">K3228025 </a> <br> (NS3 down) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228026">K3228026 </a> (oriT integration) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228069">K3228069 </a> (panS SpecRes LVL1) </li>
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| − | <li> <a href="https://parts.igem.org/Part:BBa_K3228089">K3228089 </a> (panS KanRes LVL2) </li>
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