Composite
Part:BBa_K191003:Design
Designed by: Le Thanh Tu NGUYEN Group: iGEM09_EPF-Lausanne (2009-10-11)
Promoter Lac I - RBS - LovTAP - Term
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 370
Illegal PstI site found at 627 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 370
Illegal PstI site found at 627 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 370
Illegal PstI site found at 627 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 370
Illegal PstI site found at 627 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 764
Design Notes
- Notice : 2 restriction sites of PstI in LovTAP sequence so LovTAP can't be cut with PstI.
- LovTAP was obtained by PCR and ligated into double-term plasmid ( LovTAP was cut with E and S, plasmid E and X)
- LovTAP-Term is then amplified by PCR and cut with E and X to be put into another plasmid of high-copy and only Kanamycine resistance.
- LacI was ligated into RBS
- PCR on LacI-RBS, digestion with E and S then ligated into LovTAP-Term plasmid which was cut by E and X.
Source
BBa_R0010, BBa_B0030, BBa_B0015.
Plasmid containing LovTAP gene was kindly provided by the authors of : Light-activated DNA binding in a designed allosteric protein; Strickland et al., Proceedings of the National Academy of Sciences (2008) vol. 105 (31) pp. 10709
References
Light-activated DNA binding in a designed allosteric protein; Strickland et al., Proceedings of the National Academy of Sciences (2008) vol. 105 (31) pp. 10709