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 Synechocystis PCC6803 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 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 [https://parts.igem.org/Part:BBa_J04450 device] for visual selection of posotive biobrick integration in E. coli. | + | Between biobrick prefix and suffix it has mRFP under LacI promoter (from this [https://parts.igem.org/Part:BBa_J04450 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. | 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. | ||
Revision as of 22:45, 22 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.
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
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 1135
Illegal suffix found in sequence at 2226 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1135
Illegal XhoI site found at 3689
Illegal XhoI site found at 4581 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 1135
Illegal suffix found in sequence at 2227 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2783