Composite
psb1C3_int

Part:BBa_K743008

Designed by: Bernardo Pollak, Isaac Nuñez   Group: iGEM12_UC_Chile   (2012-09-20)
Revision as of 13:46, 23 September 2012 by Juan Alamos (Talk | contribs)

psb1C3_IntKR recombination plasmid for Synechocystis PCC6803

This plasmid is intended to be used as a starting backbone for further adition of dna parts by Gibson assembly between RS1 and B0015. As synechocystis naturally undergoes double homologous recombination, any sequence between RS1 and RS2 will be introduced in to it´s 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.

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

Gel UC Chile.jpg Verification Colony PCR

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

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


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Categories
Parameters
None