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Part:BBa_K1945002:Design

Designed by: Sharon Lian   Group: iGEM16_CSU_Fort_Collins   (2016-10-14)


Homologous Recombination Intermediate for Counter-selection in Synechocystis sp. PCC 6803


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1589
    Illegal PstI site found at 305
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1589
    Illegal NheI site found at 1447
    Illegal PstI site found at 305
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1589
    Illegal BamHI site found at 2
    Illegal BamHI site found at 12
    Illegal XhoI site found at 3057
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1589
    Illegal PstI site found at 305
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1589
    Illegal PstI site found at 305
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2550


Design Notes

The counter-selection method is detailed on our wiki http://2016.igem.org/Team:CSU_Fort_Collins/Description. This piece is not in pSB1C3 and not compatible with the iGEM Registry as it is only a cloning intermediate with a very specific purpose for this strain of Synechocystis.

Please contact Christie Peebles at christie.peebles@colostate.edu for inquires about a strain of cyanobacteria already containing this counter-selection sequence.

Source

MazF is a endoribonuclease from E. coli which serves as a protein synthesis inhibitor.

References

The counter-selection homologous recombination method was developed by Cheah, Y. E., Albers, S. C., & Peebles, C. A. (2012). A novel counter-selection method for markerless genetic modification inSynechocystissp. PCC 6803. Biotechnology Progress, 29(1), 23-30. doi:10.1002/btpr.1661.