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| | + | <img src="https://static.igem.org/mediawiki/2013/4/45/Biobrick3-freiburg-2013.png" style="width:800px; margin-left:130px; margin-top:90px;"> |
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| - | <p id="h1">
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| - | BioBricks
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| - | </p>
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| - | <p id="h2">
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| - | Our BioBricks
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| - | </p>
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| - | <div id="content_parts">
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| - | <table id="tabelle">
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| - | <table border="3" frame="box">
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| - | <tr> <th> Parts </th> <th> Biobrick-Number </th> <th> Length [bp] </th> <th> Designer </th> </tr>
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| - | <tr> <td> Cas9 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150000"> BBa_K1150000 </a> </td> <td> 4101 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> VP16 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150001"> BBa_K1150001 </a> </td> <td> 372 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> KRAB </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150002"> BBa_K1150002 </a> </td> <td> 366 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> G9a </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150003"> BBa_K1150003 </a> </td> <td> 840 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> PhyB </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150004"> BBa_K1150004 </a> </td> <td> 1950 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> PIF6 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150005">BBa_K1150005 </a> </td> <td> 300 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> UVR8 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150006">BBa_K1150006 </a> </td> <td> 1110 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> COP1 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150007">BBa_K1150007 </a> </td> <td> 1023 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> NLS </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150010">BBa_K1150010 </a> </td> <td> 21 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> SV40 Promotor RFC 10 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150011">BBa_K1150011 </a> </td> <td> 344 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> BGH Terminator RFC10 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150012">BBa_K1150012 </a> </td> <td> 222 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> 3 AS Linker </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150013">BBa_K1150013 </a> </td> <td> 9 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> 10 AS Linker </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150014">BBa_K1150014 </a> </td> <td> 30 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> CMV Promotor RFC 10 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150015">BBa_K1150015 </a> </td> <td> 580 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> HA tag </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150016">BBa_K1150016 </a> </td> <td> 27 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | </table>
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| - | <td colspan="4">
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| - | <div id="content_devices">
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| - | <table id="tabelle">
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| - | <table border="3" frame="box">
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| - | <tr> <th colspan="4"> <center> Devices & others </center> </th> </tr>
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| - | <tr> <th> Description </th> <th> Biobrick-Number </th> <th> Length [bp] </th> <th> Designer </th> </tr>
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| - | <tr> <td> Cas9 with CMV promoter </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150017"> BBa_K1150017 </a> </td> <td> 5012 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Cas9 with SV40 promoter </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150018"> BBa_K1150018</a> </td> <td> 4776 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS Activator (SV40 promoter) </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150019">BBa_K1150019 </a> </td> <td> 5173 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS Activator (CMV promoter) </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150020 "> BBa_K1150020 </a> </td> <td> 5409 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS Repressor (SV40 promoter) </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150021 "> BBa_K1150021 </a> </td> <td> 5165 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS Repressor (CMV promoter) </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150022 "> BBa_K1150022 </a> </td> <td> 5401 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS Histone Modificator (SV40 promoter) </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150023"> BBa_K1150023</a></td> <td> 5641 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS Histone Modificator (CMV promoter) </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150024 "> BBa_K1150024 </a> </td> <td> 5877 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS Red Light Switch Part I - Stimulator </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150025 "> BBa_K1150025 </a> </td> <td> 5335 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS Red Light Switch Part II - Activator </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150026 "> BBa_K1150026 </a> </td> <td> 3192 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS Red Light Switch Part II - Repressor </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150027 "> BBa_K1150027 </a> </td> <td> 3184 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS Red Light Switch Part II - Histone Modificator </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150028 "> BBa_K1150028 </a> </td> <td> 3660 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS UV Light Switch Part I - Stimulator </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150029 "> BBa_K1150029 </a> </td> <td> 6147 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS UV Light Switch Part II - Activator </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150030 "> BBa_K1150030 </a> </td> <td> 2265 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS UV Light Switch Part II - Repressor </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150031 "> BBa_K1150031 </a> </td> <td> 2257 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS UV Light Switch Part II - Histone Modificator </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150032"> BBa_K1150032</a> </td> <td> 2733 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Constitutive GFP Reporter </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150033 "> BBa_K1150033 </a> </td> <td> 1711 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> dCAS RNA plasmid: "uniCAS RNaimer" </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150034 "> BBa_K1150034 </a> </td> <td> 815 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS RNaimer to EMX1 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150035 "> BBa_K1150035 </a> </td> <td> 829 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS RNaimer to target VEGF1 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150036 "> BBa_K1150036 </a> </td> <td> 829 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS RNaimer to target VEGF2 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150037 "> BBa_K1150037 </a> </td> <td> 829 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS RNaimer to target VEGF3 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150038 "> BBa_K1150038 </a> </td> <td> 829 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS RNaimer to target VEGF4 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150039"> BBa_K1150039</a> </td> <td> 829 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS RNaimer to Bla Target 2 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150040 "> BBa_K1150040 </a> </td> <td> 829 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS RNaimer to Bla Target 3 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150041 "> BBa_K1150041 </a> </td> <td> 829 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS RNaimer to Bla Target 4 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150042 "> BBa_K1150042 </a> </td> <td> 829 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS RNaimer multiple targets to VEGF 2 and VEGF 3 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150043 "> BBa_K1150043 </a> </td> <td> 1666 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS RNaimer multiple targets to Bla target 3 and 4 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150044 "> BBa_K1150044 </a> </td> <td> 1666 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS RNaimer multiple targets to VEGF 4 and bla target 2 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150045 "> BBa_K1150045 </a> </td> <td> 1666 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> uniCAS RNaimer multiple targets to Bla target 2, 3, 4 and VEGF 4 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150046"> BBa_K1150046</a> </td> <td> 3340 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Truncated CMV Cas9 Device #1 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150047 "> BBa_K1150047 </a> </td> <td> 4676 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Truncated CMV Cas9 Device #2 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150048 "> BBa_K1150048 </a> </td> <td> 4706 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Truncated CMV Cas9 Device #3 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150049 "> BBa_K1150049 </a> </td> <td> 3881 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Truncated CMV Cas9 Device #4 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150050 "> BBa_K1150050 </a> </td> <td> 3626 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Truncated CMV Cas9 Device #5 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150051 "> BBa_K1150051 </a> </td> <td> 3722 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Truncated SV40 Cas9 Device #1 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150052 "> BBa_K1150052 </a> </td> <td> 4440 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Truncated SV40 Cas9 Device #2 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150053 "> BBa_K1150053 </a> </td> <td> 4470 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Truncated SV40 Cas9 Device #3 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150054 "> BBa_K1150054 </a> </td> <td> 3645 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Truncated SV40 Cas9 Device #4 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150055 "> BBa_K1150055 </a> </td> <td> 3390 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Truncated SV40 Cas9 Device #5 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150056 "> BBa_K1150056 </a> </td> <td> 3486 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Truncated Cas9 VP16 Device #1 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150057"> BBa_K1150057</a> </td> <td> 4837 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Truncated Cas9 VP16 Device #2 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150058 "> BBa_K1150058 </a> </td> <td> 4867 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Truncated Cas9 VP16 Device #3 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150059 "> BBa_K1150059 </a> </td> <td> 4042 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Truncated Cas9 VP16 Device #4 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150060 "> BBa_K1150060 </a> </td> <td> 3787 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Truncated Cas9 VP16 Device #5 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150061 "> BBa_K1150061 </a> </td> <td> 3883 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> NLS-BGH terminator </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150062 "> BBa_K1150062 </a> </td> <td> 251 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> HA-NLS-Cas9-Linker 3 AA </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150063 "> BBa_K1150063 </a> </td> <td> 4182 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Cas9 trunc. #1 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150064 "> BBa_K1150064 </a> </td> <td> 3765 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Cas9 trunc. #2 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150065 "> BBa_K1150065 </a> </td> <td> 3795 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Cas9 trunc. #3 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150066 "> BBa_K1150066 </a> </td> <td> 2970 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Cas9 trunc. #4 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150067 "> BBa_K1150067 </a> </td> <td> 2715 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | <tr> <td> Cas9 trunc. #5 </td> <td> <a href="https://2013.igem.org/Team:Freiburg/parts/BBa_K1150068 "> BBa_K1150068 </a> </td> <td> 2811 </td> <td> Team Freiburg 2013 </td> </tr>
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| - | </table>
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| - | <div id="sharing">
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| - | <p id="h2">
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| - | Sharing of our BioBricks
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| - | </p>
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| - | </div>
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| - | <p id="h2">
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| - | Our favorite BioBricks
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| - | </p>
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| - | <p id="h2">
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| - | Improvement of an existing Registry part
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| - | </p>
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| - | <p> We decided to improve the BioBrick BBa_K128001 coding for the HA-tag – a tag which is commonly used to detect proteins. As it is cloned to all of our designed devices, we were in need of a well-functioning and easy-clonable HA-part. </p>
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| - | <p> The BBa_K128001 contains the RFC23 pre- and suffix (silver standard) which can be used for the construction of in-frame-fusion parts. In contrast to the RFC25 biobrick (which is also suitable for functional part fusions) there is no start codon and stop codon automatically included in the prefix and suffix. </p>
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| - | <p> That’s why we designed a new version of the HA-tag BBa_K128001 with an identical sequence of amino acids but with an RFC25 pre- and suffix. This has following advantages:
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| - | <ul >
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| - | <li > Cloning the HA-tag BBa_K1150016 N-terminal to other fusion parts leads to an automatic transcriptional start due to the introduced ATG in the RFC25 prefix.</li >
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| - | <li > Cloning the HA-tag BBa_K1150016 C-terminal to other fusion parts leads to an automatic transcriptional stop due to the introduced stop codon in the RFC25 suffix.</li >
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| - | </li >
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| - | </ul >
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| - | </p>
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| - | <p> As tags are often fused at the end (C- or N-terminal) of proteins, this HA-tag facilitates cloning. </p>
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| - | <table class="gelpic">
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| - | <tr>
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| - | <td> <img class="gelpic" src=" https://static.igem.org/mediawiki/parts/d/d7/HA_neu_2_Freigem_2013.png"width="100%"> </td>
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| - | </tr>
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| - | <tr>
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| - | <td <b> Fig.1: BBa_K1150016 with RFC25 prefix and suffix. </b> </td>
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| - | </tr>
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| - | </table>
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| - | <p> Before bringing this tag into the iGEM-standard we first tested its functionality via Western blot detection, see Fig.2. As it worked fine in mammalian cells, we used this sequence for our standardized constructs. Testing of our standardized devices including the BBa_K1150016 worked well and also western blot detection was possible, see Fig.3. </p>
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| - | <td> <img class="gelpic" src=" https://static.igem.org/mediawiki/parts/9/95/HA_neu_nicht_standardisiert_Freigem_2013.png"> </td>
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| - | <td <b> Fig.2: HA-tag fused to CRY2-C-RAF-CIBN-protein encoded on the pEF6/V5-His-TOPO plasmid.</b></td>
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| - | <td> <img class="gelpic" src="https://static.igem.org/mediawiki/2013/0/04/UniBAssblotfertig2.1-team_Freiburg.PNG"> </td>
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| - | <p> <td <b> Fig.3: HA-tag fused to different dcas9-fusion proteins encoded on the RFC25 pSB1C3 backbone.</b></td></p>
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