Plasmid_Backbone

Part:BBa_K260008

Designed by: Kaj Bernhardt   Group: iGEM09_BIOTEC_Dresden   (2009-10-18)

pMA-RQ-@CC2FosMB-10000bp

This plasmid backbone is based on pMA-RQ from Geneart. It has BioBrick prefix and suffix flanked on both sides by 70 bp of homology arms for insertion of the contained BioBrick into the 10000 locus of pCC2FosMB (BBa_K260001) by recombineering.

The pMA-RQ-@CC2FosMB-10000bp backbone is available with BBa_K260015 depicted below, which is composed of promoter-FRT-dhfr. PMA-RQ-P FRT dhfr 10000bp.jpg

If the PvuII fragment from BBa_K260008-BBa_K260015 is transferred by recombineering to the 10000 locus of BBa_K260001-BBa_K260016, the intervening sequence between the FRT sites of BBa_K260015 and BBa_K260016 on the BBa_K260001 backbone will be 10,000 bp exactly.

These plasmid backbones are similar and all allow recombineering-mediated transfer of the contained BioBrick (typically BBa_K260015) to the pCC2FosMB backbone BBa_K260001. They differ in the homology arms, which determine the locus of insertion into pCC2FosMB. The bp numbers denote final inter-FRT distance as described above:

BBa_K260004: pMA-CC2FosMB-00500bp

BBa_K260005: pMA-CC2FosMB-01000bp

BBa_K260006: pMA-CC2FosMB-02000bp

BBa_K260007: pMA-CC2FosMB-05000bp

BBa_K260008: pMA-RQ-CC2FosMB-10000bp


Sequence and Features


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 2527
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 2533
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2527
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 2527
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 2527
    Plasmid lacks a suffix.
    Illegal XbaI site found at 2542
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.


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Categories
Parameters
None