Translational_Unit
Part:BBa_K1465301:Design
Designed by: iGEM-Team Bielefeld 2014 Group: iGEM14_Bielefeld-CeBiTec (2014-10-06)
adhA gene from Lactococcus lactis
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 436
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
For the isolation of the adhA gene from Lactococcus lactis we first had to isolate the genomic DNA. The ahdA gene was combined with the RBS BBa_B0034 via PCR amplification. Together with the amplified backbone we performed a Gibson Assembly. We verified the correct insertion of the adhA connected with RBS in the backbone pSB1K3 via colony PCR and sequencing. Please note, that due to difficulties during the cloning process, we first did an intermediate cloning step into the pSB1K3 backbone. For that reason we performed a successful recloning in pSB1C3 which lead to the pSB1C3_adhA.
Source
- We used the gemomic DNA from Lactococcus lactis
- BBa_B0034: RBS