Coding

Part:BBa_K2560271:Design

Designed by: Memduha Muratoglu   Group: iGEM18_Marburg   (2018-10-02)


Flp recombinase

Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 747
    Illegal SpeI site found at 1065
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 747
    Illegal SpeI site found at 1065
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 747
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 747
    Illegal SpeI site found at 1065
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 747
    Illegal SpeI site found at 1065
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal EcoRI site found at 747
    Illegal SpeI site found at 1065


Design Notes

The part contains the whole coding region from the FLP recombinase gene of S. cerevisiae without the start- and stopcodon. This region is flanked by overhangs which are Phytobrick- and MoClo-compatible and by two BsaI recognition sites (Weber et al., 2011).


Source

Source of this sequence is the pBR-flp plasmid (Blokesh et al., 2012) This part was integrated into the vector BBa_K2560002 via BsmBI

References

Blokesch, M., (2012) TransFLP--a method to genetically modify Vibrio cholerae based on natural transformation and FLP-recombination. Journal of visualized experiments : JoVE.

Broach, J.R., V.R. Guarascio & M. Jayaram, (1982) Recombination within the yeast plasmid 2mu circle is site-specific. Cell 29: 227-234.

Buchholz, F., P.O. Angrand & A.F. Stewart, (1998) Improved properties of FLP recombinase evolved by cycling mutagenesis. Nature biotechnology 16: 657-662.

Sadowski, P.D., (1995) The Flp recombinase of the 2-microns plasmid of Saccharomyces cerevisiae. Progress in nucleic acid research and molecular biology 51: 53-91.

Weber E., Engler C., Gruetzner R., Werner S., Marillonnet S. (2011). A modular cloning system for standardized assembly of multigene constructs. PLoS One 6:e16765. 10.1371/journal.pone.0016765