Difference between revisions of "Part:BBa K743010"
 
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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 <i>Synechocystis PCC6803</i> chromosome and/or pSYSX endogenous plasmid while interrupting CopS gene coding sequence.
 
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 <i>Synechocystis PCC6803</i> chromosome and/or pSYSX endogenous plasmid while interrupting CopS gene coding sequence.
  
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This part has mRFP under LacI promoter (from this [https://parts.igem.org/Part:BBa_J04450 device]) between biobrick prefix and suffix for visual selection of positive biobrick integration in <i>E. coli</i>.
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This part has mRFP under LacI promoter (from this [https://parts.igem.org/Part:BBa_J04450 device]) between biobrick prefix and suffix for visual selection of biobrick integration in <i>E. coli</i>.
  
 
<i>Synechocystis PCC6803</i> undergoes double homologous recombination naturally, 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.
 
<i>Synechocystis PCC6803</i> undergoes double homologous recombination naturally, 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.

Latest revision as of 16:30, 28 September 2012

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.

This part has mRFP under LacI promoter (from this device) between biobrick prefix and suffix for visual selection of biobrick integration in E. coli.

Synechocystis PCC6803 undergoes double homologous recombination naturally, 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 environment 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