Part:BBa_K4831007:Design
P_A1lacO-1-RiboJ
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1498
Illegal SpeI site found at 1511 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1498
Illegal SpeI site found at 1511 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1498
Illegal BglII site found at 1612
Illegal XhoI site found at 1481 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1498
Illegal SpeI site found at 1511 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1498
Illegal SpeI site found at 1511 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
Added RiboJ which is a self-cleaving ribonuclease element. RiboJ cleaves the part in mRNA between 5' UTR and the starting codon so that the mRNA molecules are always the same size and cutting the extra sequence also makes the mRNA less stable, which decreases the unspesific background when inducing the activity or when measuring the uninduced signal.
Source
http://dx.doi.org/10.1371/journal.pone.0050470
References
Guerrero, F., Carbonell, V., Cossu, M., Correddu, D. and Jones. P. R. (2012) Ethylene synthesis and regulated expression of recombinant protein in Synechocystis sp. PCC 6803. PLoS ONE 7(11):e50470.http://dx.doi.org/10.1371/journal.pone.0050470
Thiel, K., Mulaku, E., Dandapani, H., Nagy, C., Aro, E-M., Kallio, P. (2018) Translation efficiency of heterologous proteins is significantly affected by the genetic context of RBS sequences in engineered cyanobacterium Synechocystis sp. PCC 6803. Microbial Cell Factories 17(1):34. https://doi.org/10.1186/s12934-018-0882-2
Thiel K, Patrikainen P, Nagy C, Fitzpatrick D, Pope N, Aro EM, Kallio P. (2019) Redirecting photosynthetic electron flux in the cyanobacterium Synechocystis sp. PCC 6803 by the deletion of flavodiiron protein Flv3. Microbial Cell Factories, 18: 189. https://doi.org/10.1186/s12934-019-1238-2
Nagy, C., Thiel, K., Mulaku, E., Mustila, H., Tamagnini, P., Aro, E-M, Pacheco, C. C., Kallio, P. (2021) Comparison of alternative integration sites in the chromosome and the native plasmids of the cyanobacterium Synechocystis sp. PCC 6803 in respect to expression efficiency and copy number. Microbial Cell Factories 20, 130. https://doi.org/10.1186/s12934-021-01622-2