Difference between revisions of "Part:BBa K2333414"
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<partinfo>BBa_K2333414 short</partinfo> | <partinfo>BBa_K2333414 short</partinfo> | ||
| − | mScarlet | + | This part is contained in a suite of protein degradation tagged mScarlet reporters under the control of the strong constitutive promoter J23100. These parts, in combination with inducible mf-Lon protease constructs, allowed us to characterize the degradation properties of each protein degradation tag (pdt) on a plasmid-based system. We successfully demonstrated distinct levels of protein degradation by each of the 6 pdt’s, and mScarlet reporters have been codon-optomized for E. coli and feature a double stop codon for enhanced efficiency. This specific part is a tagless control construct (J23100 mScarlet with no pdt) which can be used as a comparison against protein degradation for parts with pdt's. |
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===Usage and Biology=== | ===Usage and Biology=== | ||
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| + | This part contains mScarlet-I with pdt-A under the control of the constitutive promoter J23100. Protein degradation tag A is the strongest of the 6 protein degradation tags that William and Mary 2017 characterized, and is associated with the E. Coli orthogonal protease mf-Lon (<partinfo>Bba_K2333011</partinfo>). This part also contains a double stop codon and <partinfo>Bba_B0015</partinfo> (double terminator) in the William and Mary iGEM Universal Nucleotide Sequences (UNS) format. This enables easy cloning with Gibson Assembly, as UNS primers are designed for easy PCRs and high yield Gibson Assembly. When used in combination with inducible mf-Lon protease constructs, William and Mary 2017 was able to characterize the degradation properties of protein degradation tag A on a plasmid-based system. This is a part of the first experimentally-demonstrated system that allows future iGEM teams to access modular, predictive control over the temporal dynamics of their circuits by swapping parts at the genetic sequence level. | ||
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Revision as of 17:51, 29 October 2017
UNS J23100 mScarlet-I pdt A
This part is contained in a suite of protein degradation tagged mScarlet reporters under the control of the strong constitutive promoter J23100. These parts, in combination with inducible mf-Lon protease constructs, allowed us to characterize the degradation properties of each protein degradation tag (pdt) on a plasmid-based system. We successfully demonstrated distinct levels of protein degradation by each of the 6 pdt’s, and mScarlet reporters have been codon-optomized for E. coli and feature a double stop codon for enhanced efficiency. This specific part is a tagless control construct (J23100 mScarlet with no pdt) which can be used as a comparison against protein degradation for parts with pdt's.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 47
Illegal NheI site found at 70
Illegal NotI site found at 605 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]