Coding
Part:BBa_K525562:Design
Designed by: Anna Drong Group: iGEM11_Bielefeld-Germany (2011-10-28)
Fusion protein of NADP+ Oxidoreductase and BisdA and BisdB with middle strong promoter,RBS
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1411
Illegal BamHI site found at 2149 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 65
Illegal AgeI site found at 2402 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 746
Design Notes
- Fusion protein of FNR, BisdA and BisdB behind medium strong constitutive promoter to avoid overexpression which can leed to misfolding as [http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2672.2008.03843.x/full Sasaki et al. (2008)] reported.
- Fusion protein in and assembled via Freiburg BioBrick assembly standard (RFC 25) leaving AgeI and NgoMIV restriction sites
Source
- bisdA and bisdB originated in Sphingomonas bisphenolicum AO1
- FNR originated in Escherichia coli TOP10
- Anderson promoter BBa_J23110 and RBS BBa_B0034 from parts.igem
- All subparts (except the FNR) can be found in the kit plates
References
Sasaki M, Tsuchido T, Matsumura Y (2008) Molecular cloning and characterization of cytochrome P450 and ferredoxin genes involved in bisphenol A degradation in Sphingomonas bisphenolicum strain AO1, [http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2672.2008.03843.x/full J Appl Microbiol 105(4):1158-1169].