Difference between revisions of "Part:BBa K1758312:Design"
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+ | <partinfo>BBa_K1758312 SequenceAndFeatures</partinfo> | ||
+ | ===Design Notes=== | ||
+ | <html> | ||
+ | For changing the repressor from <i>RFP</i> to sfGFP and adding the 5’UTR we designed primers for Gibson Assembly to ligate the <i>sfGFP</i> with the <i>crP</i>. For this aim we designed four primers for the generation of homologous overlaps between <i>5’-UTR-sfGFP</i> and <i>crP</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>crProm </i> plasmid (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1758311 " target="_blank"> BBa_K1758311 </a>) : | ||
+ | <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> | ||
+ | <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> | ||
+ | <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> | ||
+ | <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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Latest revision as of 02:54, 19 September 2015
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 136
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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 crProm plasmid ( BBa_K1758311 ) :
CGGCATCAGCACCTTGTC
CTTTTTTTTTTTTAAAGTTAAACAAAATTATTGTCGTAGATGTTACTACAATAATGAG
TATACGCAAGGCGACAAG
GTTTAACTTTAAAAAAAAAAAAGAAGGAGAATAATCTCTGCGTAAAGGCGAAGAGCTGTTCAC
Source
Biobrick modified after K1058007