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
    COMPATIBLE WITH RFC[1000]


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