Coding

Part:BBa_K2560270:Design

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


SXT operon


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 496
    Illegal EcoRI site found at 2005
    Illegal PstI site found at 754
    Illegal PstI site found at 1001
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 496
    Illegal EcoRI site found at 2005
    Illegal PstI site found at 754
    Illegal PstI site found at 1001
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 496
    Illegal EcoRI site found at 2005
    Illegal BamHI site found at 2066
    Illegal XhoI site found at 3318
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 496
    Illegal EcoRI site found at 2005
    Illegal PstI site found at 754
    Illegal PstI site found at 1001
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 496
    Illegal EcoRI site found at 2005
    Illegal PstI site found at 754
    Illegal PstI site found at 1001
    Illegal AgeI site found at 574
    Illegal AgeI site found at 2231
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 2248


Design Notes

The part contains the whole coding region from an SXT operon consisting of SXT-Exo, SXT-Beta from V. cholerae and lambda-Gam from E. coli under the control of an arabinose promoter. This region is flanked by overhangs which are Phytobrick- and MoClo-compatible and by two BsaI recognition sites (Weber et al., 2011). But the coding region contains EcoRI and PstI restriction sites.

Source

The source of the SXT operon was a plasmid, encoding the full sequence. This part was integrated into the vector BBa_K2560002 via BsmBI

References

Church, G.M., Gold, M.A., Ostrov, N., Lee, H.H., (2017) Recombineering in Vibrio natriegens. bioRxiv.

Murphy, K.C., (2016) lambda Recombination and Recombineering. EcoSal Plus 7.

Sharan, S.K., L.C. Thomason, S.G. Kuznetsov & D.L. Court, (2009) Recombineering: a homologous recombination-based method of genetic engineering. Nature protocols 4: 206-223.

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