Difference between revisions of "Part:BBa K743008"
 
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<partinfo>BBa_K743008 short</partinfo>
 
<partinfo>BBa_K743008 short</partinfo>
  
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This plasmid allows the integration of a double terminator followed by a reversed Kanamycin resistance casette in to Synechocystis chromosome, making possible selection in this cyanobacteria. It has a Chloramphenicol resistance casette for selection in E.coli
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This plasmid is intended to be used as a starting backbone for further addition of DNA parts by Gibson assembly between RS1 and B0015. As <i>Synechocystis PCC6803</i> undergoes double homologous recombination naturally, any sequence between RS1 and RS2 will be introduced into its chromosome along with a KanR gene for selection in cyanobacteria or in <i>E. coli</i>.
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The use of [https://parts.igem.org/Part:BBa_K743000 BBa_K743000] and [https://parts.igem.org/Part:BBa_K743001 BBa_K743001] as recombination sites has no deleterious phenotypic effects on the cells.
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The plasmid is intended to be used as a starting backbone for further adition of dna parts by Gibson assembly.  
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It was used by [http://2012.igem.org/Team:UC_Chile UC_Chile 2012 team] to succesfully transform Synechocystis
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[[Image:gelC4_UC_Chile.jpg|400px]]
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The plasmid backbone also has a Chloramphenicol resistance casette for selection in <i>E. coli</i> only.
  
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[[Image:Example.jpg]]
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The use of [https://parts.igem.org/Part:BBa_K743000 BBa_K743000] and [https://parts.igem.org/Part:BBa_K743001 BBa_K743001] as recombination sites has no deleterious phenotypic effects on the cells.
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This part was used by [http://2012.igem.org/Team:UC_Chile UC_Chile 2012 team] to succesfully transform <i>Synechocystis PCC6803</i>.
  
 
<!-- Add more about the biology of this part here
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K743008 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K743008 SequenceAndFeatures</partinfo>
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[[Image:gel_UC_Chile.jpg|400px]]
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'''Verification by Colony PCR'''
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[[Image:gel_uc_chile.jpg|400px]]
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'''psb1C3_IntK plasmid with BBa_K743008 digested with EcoR1 and Pst1'''
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Latest revision as of 19:46, 24 September 2012

psb1C3_IntKR recombination plasmid for Synechocystis PCC6803

This plasmid is intended to be used as a starting backbone for further addition of DNA parts by Gibson assembly between RS1 and B0015. As Synechocystis PCC6803 undergoes double homologous recombination naturally, any sequence between RS1 and RS2 will be introduced into its chromosome along with a KanR gene for selection in cyanobacteria or in E. coli.

The plasmid backbone also has a Chloramphenicol resistance casette for selection in E. coli only.

The use of BBa_K743000 and BBa_K743001 as recombination sites has no deleterious phenotypic effects on the cells.

This part was used by [http://2012.igem.org/Team:UC_Chile UC_Chile 2012 team] to succesfully transform Synechocystis PCC6803.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1758
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1758
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1758
    Illegal XhoI site found at 2255
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1758
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1758
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 511

Gel UC Chile.jpg Verification by Colony PCR

Gel uc chile.jpg psb1C3_IntK plasmid with BBa_K743008 digested with EcoR1 and Pst1