Difference between revisions of "Part:BBa K1758325:Design"
 
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===Design Notes===
 
===Design Notes===
 
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<p> BioBrick upstream insertion of the Biobrick <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_ I719005" target="_blank">BBa_I719005 </a> into the BioBrick <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1758323" target="_blank">BBa_K1758323</a>. </p>
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For changing the repressor from <i>RFP</i> to <i>sfGFP</i> and adding the 5’UTR  we designed primers for Gibson Assembly to ligate the <i>sfGFP</i> with the <i>copAP</i>. For this aim we designed four primers for the generation of homologous overlaps between 5’-UTR-<i>sfGFP</i> and <i>copAP</i> used a template with <i>sfGFP</i> like (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1758102 " target="_blank"> BBa_K1758102</a>) and <i>copAP</i> plasmid (<a BBa_K1758321 href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1758321 " target="_blank"> BBa_K1758321</a>) :
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<p><a href="http://2015.igem.org/Team:Bielefeld-CeBiTec/Primers#cm_fwd" target="_blank"> <a href="http://2015.igem.org/Team:Bielefeld-CeBiTec/Primers#cm_fwd" target="_blank">cm_fwd</a></p> <p>CGGCATCAGCACCTTGTC </p>
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<p> <a href="http://2015.igem.org/Team:Bielefeld-CeBiTec/Primers#copAP_UTR_rev " target="_blank"> <a href="http://2015.igem.org/Team:Bielefeld-CeBiTec/Primers#copAP_UTR_rev" target="_blank"> copAP_UTR_rev</a></p> CCTTCTTTTTTTTTTTTAAAGTTAAACAAAATTATTCTAGTACTTTAAGACAGTTTTG<p>
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<p> <a href="http://2015.igem.org/Team:Bielefeld-CeBiTec/Primers#cm_rev" target="_blank"> <a href="http://2015.igem.org/Team:Bielefeld-CeBiTec/Primers#cm_rev" target="_blank">cm_rev</a></p> <p> TATACGCAAGGCGACAAG </p>
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<p><a href="http://2015.igem.org/Team:Bielefeld-CeBiTec/Primers#UTR_mod_fwd" target="_blank"> <a href="http://2015.igem.org/Team:Bielefeld-CeBiTec/Primers#UTR_mod_fwd" target="_blank"> UTR_mod_fwd</a> </p> <p> GTTTAACTTTAAAAAAAAAAAAGAAGGAGAATAATCTCTGCGTAAAGGCGAAGAGCTGTTCAC</p>
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===Source===
 
===Source===
 
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===References===
 
===References===

Latest revision as of 02:25, 19 September 2015

Copper responsive promoter with T7-promoter and UTR-sfGFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 29
    Illegal SapI.rc site found at 179


Design Notes

For changing the repressor from RFP to sfGFP and adding the 5’UTR we designed primers for Gibson Assembly to ligate the sfGFP with the copAP. For this aim we designed four primers for the generation of homologous overlaps between 5’-UTR-sfGFP and copAP used a template with sfGFP like ( BBa_K1758102) and copAP plasmid ( BBa_K1758321) :

cm_fwd

CGGCATCAGCACCTTGTC

copAP_UTR_rev

CCTTCTTTTTTTTTTTTAAAGTTAAACAAAATTATTCTAGTACTTTAAGACAGTTTTG

cm_rev

TATACGCAAGGCGACAAG

UTR_mod_fwd

GTTTAACTTTAAAAAAAAAAAAGAAGGAGAATAATCTCTGCGTAAAGGCGAAGAGCTGTTCAC


Source

References