Difference between revisions of "Part:BBa K1758312:Design"

 
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<partinfo>BBa_K1758312 SequenceAndFeatures</partinfo>
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===Design Notes===
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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 crP. For this aim we designed four primers for the generation of homologous overlaps between 5’-UTR-sfGFP and crP used a template with sfGFP like (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1758102 " target="_blank"> BBa_K1758102</a>) and crP plasmid (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1758311 " target="_blank"> BBa_K1758311 </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#CrProm_UTR_rev" target="_blank"> <a href="http://2015.igem.org/Team:Bielefeld-CeBiTec/Primers#CrProm_UTR_rev" target="_blank"> CrProm_UTR_rev </a></p>CTTTTTTTTTTTTAAAGTTAAACAAAATTATTGTCGTAGATGTTACTACAATAATGAG<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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<partinfo>BBa_K1758312 short</partinfo>
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<partinfo>BBa_K1758312 SequenceAndFeatures</partinfo>
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===Design Notes===
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Revision as of 02:14, 19 September 2015


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 136
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 229

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 crP. For this aim we designed four primers for the generation of homologous overlaps between 5’-UTR-sfGFP and crP used a template with sfGFP like ( BBa_K1758102) and crP plasmid ( BBa_K1758311 ) :

cm_fwd

CGGCATCAGCACCTTGTC

CrProm_UTR_rev

CTTTTTTTTTTTTAAAGTTAAACAAAATTATTGTCGTAGATGTTACTACAATAATGAG

cm_rev

TATACGCAAGGCGACAAG

UTR_mod_fwd

GTTTAACTTTAAAAAAAAAAAAGAAGGAGAATAATCTCTGCGTAAAGGCGAAGAGCTGTTCAC


Source

Biobrick modified after K1058007

References