Difference between revisions of "Part:BBa K1758325:Design"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K1758325 short</partinfo> | <partinfo>BBa_K1758325 short</partinfo> | ||
<partinfo>BBa_K1758325 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1758325 SequenceAndFeatures</partinfo> | ||
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===Design Notes=== | ===Design Notes=== | ||
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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>) : | ||
+ | <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#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> | ||
+ | <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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===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:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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) :
CGGCATCAGCACCTTGTC
CCTTCTTTTTTTTTTTTAAAGTTAAACAAAATTATTCTAGTACTTTAAGACAGTTTTG
TATACGCAAGGCGACAAG
GTTTAACTTTAAAAAAAAAAAAGAAGGAGAATAATCTCTGCGTAAAGGCGAAGAGCTGTTCAC
Source