Part:BBa_K4811019
PT7-TMS(Kan1)-TT7
This is a transcriptional unit for a tRNA mimicking structure (TMS), using the T7 promoter and terminator. In E. coli BL21(DE3), the T7 polymerase is under LacI control, meaning it will be expressed by IPTG, leading to expression of the TMS RNA. The TMS is a novel piece of synthetically designed RNA, which folds in the same manner as tRNA, developed by Paul et al. It is a trans-encoded genetic switch, which binds to the Ribosome Binding Site (RBS) BBa_K4811001, repressing translation. The binding regions are flanking the RBS, and no binding happens in the RBS consensus sequence. This means that there are very few limitations on both the possible TMSs and repressible RBSs which could be engineered. Also, the D-loop of the TMS can be exchanged for additional functionality, such as an aptamer, making the TMS responsive to a ligand.
In this particular part, the D-loop has been exchanged for a Kanamycin-sensitive aptamer.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 43
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 43
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 43
Illegal XhoI site found at 50
Illegal XhoI site found at 86 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 43
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 43
- 1000COMPATIBLE WITH RFC[1000]
None |