Composite
psb1C3_Int

Part:BBa_K743010

Designed by: Bernardo Pollak y Simon Alamos   Group: iGEM12_UC_Chile   (2012-09-22)
Revision as of 22:45, 22 September 2012 by Juan Alamos (Talk | contribs)

psb1C3_IntS recombination plasmid with RFP

psb1C3_IntS recombination plasmid has biobrick prefix and suffix sequences flanked by recombination sites, therefore it is designed to integrate any desired biobricks or other sequences into Synechocystis PCC6803 chromosome and/or pSYSX endogenous plasmid while interrupting CopS gene coding sequence.

Between biobrick prefix and suffix it has mRFP under LacI promoter (from this device) for visual selection of posotive biobrick integration in E. coli.

Synechocystis naturally undergoes double homologous recombination so the use of recombination sites flanking prefix and suffix were designed with complete homology to CopS gene, present in the chromosome and in a endogenous plasmid. As CopS is essential for copper stress tolerance in this bacteria, by using this plasmid a succeptible strain incapable of thriving in the enviroment is created (see references for more information).


Sequence and Features


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


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