Composite
psb1C3_Int

Part:BBa_K743010:Design

Designed by: Bernardo Pollak y Simon Alamos   Group: iGEM12_UC_Chile   (2012-09-22)
Revision as of 21:49, 22 September 2012 by Juan Alamos (Talk | contribs) (→‎Source)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

psb1C3_IntS recombination plasmid with RFP


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 1135
    Illegal suffix found in sequence at 2226
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1135
    Illegal SpeI site found at 2227
    Illegal PstI site found at 2241
    Illegal NotI site found at 1141
    Illegal NotI site found at 2234
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1135
    Illegal XhoI site found at 3689
    Illegal XhoI site found at 4581
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 1135
    Illegal suffix found in sequence at 2227
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 1135
    Illegal XbaI site found at 1150
    Illegal SpeI site found at 2227
    Illegal PstI site found at 2241
    Illegal NgoMIV site found at 867
    Illegal AgeI site found at 1937
    Illegal AgeI site found at 2049
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2783


Design Notes

The recombination sites are just 450bp long so that complete homology with any version of the CopS gene is achieved (i.e chromosomic and plasmidial versions) Though according to literature 1300 bp is the optimum size for recombination sites in Synechocystis, 450 bp yields reasonably high transformation efficiency.


Source

Recombination sites were PCR amplified from Synechocystis PCC6803 DNA. All the sequences between recombination sites are from psb4K5. The rest of the plasmid comes from psb1C3 sequence.

All the plasmid was built by Gibson assembly.

References

Giner-Lamia, J., Lopez-Maury, L., Reyes, J. C., & Florencio, F. J. (2012). The CopRS two-component system is responsible for resistance to copper in the cyanobacterium Synechocystis sp. PCC 6803. Plant physiology, 159(August), 1806-1818.