Part:BBa_K1806002
T7 lacO pelB RFP 6xHis
The part has been designed as a backbone part to integrate the T7 lacO pelB + 6xHis system to a targeted sequence. The RFP has BamH1 and Xho1 restriction sites to be cut out and replaced by the targeted sequence. T7 lacO allows the system to be inducible by iPTG and the pelB signal peptide allows the cells to excrete the translated proteins placed in the following sequence. The 6xHis is present for protein analysis.
Cloning
The system was ligated with the pSB1C3 cloning vector and then transformated to be verified with Colony PCR. The RFP present in the iGEM Registry contains the Hind3 enzyme restriction site unlike the RFP sequence present in this part. Hence, the part was cut with the EcoR1 and Hind3 restriction enzymes.
Tagged in the gel runs below as G-Block 4, it can be seen that the gel runs yielded accurate base length.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2
Illegal BamHI site found at 196
Illegal XhoI site found at 881 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 163
Illegal AgeI site found at 757
Illegal AgeI site found at 869 - 1000COMPATIBLE WITH RFC[1000]
| None |