Part:BBa_K1789000:Design
IaaM
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 402
Illegal BamHI site found at 1347 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 109
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The reason why er designed this part is that we needed IAA producing pathway as a reporter in our system, and the IAAM and IAAH CDS needed to be fused with the coding sequence of the TAL effector we engineered. So we used the part BBa_K515100, which contains the CDS of IaaM and IaaH, and used PCR to eliminate the RBS sequence from the 5' region of the CDS.
The PCR was performed with Premix EX Taq by TAKARA under the protocol recommended by the users manual.
F-Prime: 5’- GGAATTCGCGGCCGCTTCTAGAGATGTTTGGACCGG-3’
R-Prime: 5’- GCGGCGGACTAGTCTTATTAGTCCCCCAGCG -3’
The PCR product was purified and cut by EcoRI and SpeI, then ligated with PSB1C3 vector
Source
This part is gained from BBa_K515100 by PCR. The IAA producing pathway originates from Pseudomonas savastanoi.