Composite
Part:BBa_K3110010:Design
Designed by: Mrugank Dake Group: iGEM19_IISER_Tirupati (2019-10-14)
Weak Promoter Strong RBS lldD
Assembly Compatibility:
- 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 NgoMIV site found at 1202
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 639
Design Notes
lldD was codon optimized to meet synthesis requirements and to remove the illegal EcoRI site present in the genomic lldD.
Source
The lldD Synthesized from Twist Bioscience. Specific Promoter and RBS with D flank and Terminator with D flank was ordered from IDT and all the 3 parts were merged together using SOEing (Spliced Extension Overlapped) PCR.
References
Aguilera, L., Campos, E., Gimenez, R., Badia, J., Aguilar, J., & Baldoma, L. (2008). Dual Role of LldR in Regulation of the lldPRD Operon, Involved in L-Lactate Metabolism in Escherichia coli. Journal of Bacteriology, 190(8), 2997–3005. doi:10.1128/jb.02013-07