Difference between revisions of "Part:BBa K4365021"
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* codon optimized turboRFP [https://parts.igem.org/Part:BBa_K4365020 K4365020] | * codon optimized turboRFP [https://parts.igem.org/Part:BBa_K4365020 K4365020] | ||
* terminator tCYC1 [https://parts.igem.org/Part:BBa_K849009 K849009] | * terminator tCYC1 [https://parts.igem.org/Part:BBa_K849009 K849009] | ||
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| + | [[BBa K4365021 schematic.png|500px|center|thumb|Figure 1: illustration of SP-SUMO system for secretion and scar-less protein production in yeast. The SP-SUMO is attached to the cargo protein turboRFP as this was the construct we tested during our experiments. Created with Biorender.com.]] | ||
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<br><span class='h3bb'>Sequence and Features</span> | <br><span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K4365021 SequenceAndFeatures</partinfo> | <partinfo>BBa_K4365021 SequenceAndFeatures</partinfo> | ||
Revision as of 08:22, 11 October 2022
SP-SUMO-turboRFP
SP-SUMO-tRFP is a reporter device which could be used to test protein secretion in yeast. Moreover, if tRFP is exchanged for the protein of interest, after protein purification with His-tag you can scarlessly remove the tag from the protein with a SUMO protease. This part is optimized for expression in Saccharomyces cerevisiae.
SP-SUMO-tRFP consists of:
- FIG1 promoter K4365000
- signal peptide (alpha mating factor signal peptide) K4365019
- glycine linker K4365005
- 6x His-Tag K3033006
- SUMO K4365004
- codon optimized turboRFP K4365020
- terminator tCYC1 K849009
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 1001
Illegal PstI site found at 1036
Illegal PstI site found at 1335 - 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 1001
Illegal PstI site found at 1036
Illegal PstI site found at 1335 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2168
Illegal BamHI site found at 1007 - 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 1001
Illegal PstI site found at 1036
Illegal PstI site found at 1335 - 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 1001
Illegal PstI site found at 1036
Illegal PstI site found at 1335
Illegal NgoMIV site found at 172 - 1000COMPATIBLE WITH RFC[1000]