Difference between revisions of "Part:BBa K2671000"
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<partinfo>BBa_K2671000 short</partinfo> | <partinfo>BBa_K2671000 short</partinfo> | ||
− | Liu iGEM2017s plasmid (BBa_K2474000) with an improvement. A stop codon was mutated behind mNeonGreen (mNG). The original part was mNG fused to | + | Liu iGEM2017s plasmid (BBa_K2474000) with an improvement. A stop codon was mutated behind mNeonGreen (mNG). The original part was mNG fused to Aβ1-42, where the fusion protein had its stop codon after Aβ1-42. Another mutation was made in the GS-linker sequence where a SpeI-site was added and the sequence for Aβ1-42 was cleaved off. With these improvements mNG can now be used as a reporter in a fully functional operon with the pBAD-AraC inducible system. |
===Usage and Biology=== | ===Usage and Biology=== | ||
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Verification | Verification | ||
As seen in fig 1. the improved version has a much stronger color, which indicates that the Aß1-42 fusion was successfully terminated. And as seen in fig 2. the sequencing results confirms this. | As seen in fig 1. the improved version has a much stronger color, which indicates that the Aß1-42 fusion was successfully terminated. And as seen in fig 2. the sequencing results confirms this. | ||
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+ | [[File:T--Linkoping_Sweden--improveseq1.png|200px|thumb|left|Figure 2. Sequencing results analyzed in benchling, annotations added for clarity. The sequencing data verifies a successful removal of Aβ1-42 and addition of a stop codon. Sequencing was done with the VR primer.]] | ||
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Revision as of 14:46, 15 October 2018
AraC-pBAD-mNG
Liu iGEM2017s plasmid (BBa_K2474000) with an improvement. A stop codon was mutated behind mNeonGreen (mNG). The original part was mNG fused to Aβ1-42, where the fusion protein had its stop codon after Aβ1-42. Another mutation was made in the GS-linker sequence where a SpeI-site was added and the sequence for Aβ1-42 was cleaved off. With these improvements mNG can now be used as a reporter in a fully functional operon with the pBAD-AraC inducible system.
Usage and Biology
Verification As seen in fig 1. the improved version has a much stronger color, which indicates that the Aß1-42 fusion was successfully terminated. And as seen in fig 2. the sequencing results confirms this.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1267
Illegal BamHI site found at 1144 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961