Plasmid_Backbone

Part:BBa_J153000:Design

Designed by: Daniel Camsund   Group: Lindblad Lab   (2012-02-02)
Revision as of 16:10, 2 February 2012 by Registry (Talk | contribs)

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Broad-host-range shuttle vector pPMQAK1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 7676
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 7682
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 7676
    Illegal XhoI site found at 6569
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 7676
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 7676
    Plasmid lacks a suffix.
    Illegal XbaI site found at 7691
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal AgeI site found at 1061
    Illegal AgeI site found at 1462
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 6715


Design Notes

As cloning by plasmid preparation and subsequent digestion/ligation was problematic due to low copy number and difficulties in digesting the RSF1010-derived part from pAWG1.1, pPMQAK1 was made using PCR and subsequent digestion/ligation.



Source

pPMQAK1 was constructed by ligating a PCR product from the pAWG1.1 plasmid, corresponding to the RSF1010 replicon, with a PCR product from the pSB1AK3 plasmid, corresponding to the BioBrick cloning site with flanking functional sequences plus the ampicillin and kanamycin resistance cassettes.

References