Difference between revisions of "Part:BBa K4437503:Design"
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This sequence was designed as a positive control for experiments comparing RBS strength and resulting FLAG-phasin-HlyA expression levels, with the purpose of improving the expression system to increase phasin yields. As such, this part was designed with the same RBS type as the original BBa_K2260002, to compare the relative expression levels of the other variable RBS types when combined with the FLAG-phasin-HlyA sequence. | This sequence was designed as a positive control for experiments comparing RBS strength and resulting FLAG-phasin-HlyA expression levels, with the purpose of improving the expression system to increase phasin yields. As such, this part was designed with the same RBS type as the original BBa_K2260002, to compare the relative expression levels of the other variable RBS types when combined with the FLAG-phasin-HlyA sequence. | ||
| − | + | The restriction enzyme cut sites for BstAPI and BstBI were also added, making the part compatible for digestion and ligation into the Tokyo Tech 2012 team’s BBa_K934001 plasmid for PHB expression. | |
===Source=== | ===Source=== | ||
Latest revision as of 09:18, 12 October 2022
B0034-FLAG-phasin-HlyA (E. coli)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
This sequence was designed as a positive control for experiments comparing RBS strength and resulting FLAG-phasin-HlyA expression levels, with the purpose of improving the expression system to increase phasin yields. As such, this part was designed with the same RBS type as the original BBa_K2260002, to compare the relative expression levels of the other variable RBS types when combined with the FLAG-phasin-HlyA sequence.
The restriction enzyme cut sites for BstAPI and BstBI were also added, making the part compatible for digestion and ligation into the Tokyo Tech 2012 team’s BBa_K934001 plasmid for PHB expression.
Source
This part is based on the BBa_K2260002 part from the University of Calgary's 2017 iGEM team. The phasin (PhaP) gene is from R. eutropha, the HlyA tag is from E. coli, and the T7 promoter is from the T7 bacteriophage.