Difference between revisions of "Part:BBa J153000:Design"

 
(→‎References)
 
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_J153000 short</partinfo>
 
<partinfo>BBa_J153000 short</partinfo>
Line 17: Line 16:
  
 
===References===
 
===References===
 +
 +
 +
Huang, H.H., Camsund, D., Lindblad, P. and Heidorn, T. (2010) Design and characterization of molecular tools for a Synthetic Biology approach towards developing cyanobacterial biotechnology. Nucleic Acids Res, 38, 2577-2593.

Latest revision as of 17:29, 2 February 2012

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

Huang, H.H., Camsund, D., Lindblad, P. and Heidorn, T. (2010) Design and characterization of molecular tools for a Synthetic Biology approach towards developing cyanobacterial biotechnology. Nucleic Acids Res, 38, 2577-2593.