Difference between revisions of "Part:BBa K2956000:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | This part is a composite part consisting of BBa_J23100 promoter, BBa_B0034 ribosome binding sequence, and BBa_K2956000 coding region for flavone synthase (FNS). These parts were assembled together to allow for constitutive expression of the FNS gene which converts naringenin to apigenin. When inserted into a vector plasmid and expressed in E. coli, naringenin was shown to be converted at a rate that was positively correlated to cell growth, demonstrating the efficacy of this system. | + | This part is a composite part consisting of BBa_J23100 promoter, BBa_B0034 ribosome binding sequence, and BBa_K2956000 coding region for flavone synthase (FNS). The FNS gene comes from Petroselinum crispum. These parts were assembled together to allow for constitutive expression of the FNS gene which converts naringenin to apigenin. When inserted into a vector plasmid and expressed in E. coli, naringenin was shown to be converted at a rate that was positively correlated to cell growth, demonstrating the efficacy of this system. |
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| + | Silent mutations were introduced to BBa_K2956000 to remove illegal sites, allowing RFC[10] compatibility. | ||
===Source=== | ===Source=== | ||
Latest revision as of 16:13, 21 October 2019
RBS-FNS under J23100 Promoter
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 794
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
This part is a composite part consisting of BBa_J23100 promoter, BBa_B0034 ribosome binding sequence, and BBa_K2956000 coding region for flavone synthase (FNS). The FNS gene comes from Petroselinum crispum. These parts were assembled together to allow for constitutive expression of the FNS gene which converts naringenin to apigenin. When inserted into a vector plasmid and expressed in E. coli, naringenin was shown to be converted at a rate that was positively correlated to cell growth, demonstrating the efficacy of this system.
Silent mutations were introduced to BBa_K2956000 to remove illegal sites, allowing RFC[10] compatibility.
Source
The DNA sequences for the basic parts that are included in this system were obtained from the iGEM registry. The J23100 promoter, part BBa_J23100, was created by iGEM 2006 Berkeley and widely used in other iGEM projects for its high strength. The ribosome binding site, part BBa_B0034, was also chosen because it had been highly characterized. The coding region for flavone synthase, BBa_K2819001, was taken from Petroselinum crispum and codon optimized.