Part:BBa_K4390044
Lambda cI controlled Chloramphenicol resistance
This part is not compatible with BioBrick RFC10 assembly but is compatible with the iGEM Type IIS Part standard which is also accepted by iGEM.
This is a level 1 part formed by assembly of the following level 0 parts:
| Promoter | R0051 |
| RBS | B0034 |
| N-O-C part | J31005 |
| Terminator | K4390001 |
Usage and Biology
The promoter of this designed part is able to be regulated by Lambda cI, which is a transcriptional repressor which allows Lambda phage to establish and maintain latency after infect E. coli. It regulates the entry of lytic cycle by repressing the lytic promoters (Johnson, A. D. et al., 1979). This Lambda cI sequence was codon optimised for expression in E. coli K12, and was be used in Seamless Enrichment of Ligand-Inducible Sensors (SELIS) as the repressor (d’Oelsnitz, S. et al., 2022). CmR gene encodes protein which is able to induce chloramphenicol resistance by triggering putative efflux pump (Nilsen, I. W. et al., 1996).
In this designed part, the CmR gene will be expressed under regulation of Lambda cI regulated promoter, thus CmR gene will only be expressed when there are no Lambda cI present in the environment. In iGEM22_Edinburgh-UHAS_Ghana SELIS design, the expressed CmR gene will trigger chloramphenicol resistance to the bacteria which can be used to select the bacteria that contains the desired circuit. Since Lambda cI is normally been regulated, colonies that expressing this part will be able to survive in chloramphenicol plates, whereas the colonies that does not contain CmR gene will be killed. For further information please refer to any of SELIS arsR evolution construct, SELIS merR evolution construct, SELIS mutated merR evolution construct and SELIS pbrR evolution construct.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
References
D'Oelsnitz, S. et al., (2022) Using fungible biosensors to evolve improved alkaloid biosyntheses. Nature chemical biology. 18 (9), 981–989.
Johnson, A. D. et al. (1979) Interactions between DNA-Bound Repressors Govern Regulation by the $\lambda $ Phage Repressor. Proceedings of the National Academy of Sciences - PNAS. 76 (10), 5061–5065.
Nilsen, I. W. et al. (1996) Isolation of cmr, a novel Escherichia coli chloramphenicol resistance gene encoding a putative efflux pump. Journal of Bacteriology. 178 (11), 3188–3193.
| None |