Difference between revisions of "Part:BBa K260010"

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K260010 short</partinfo>
 
<partinfo>BBa_K260010 short</partinfo>
  
 
DESCRIPTION:
 
DESCRIPTION:
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This plasmid backbone is based on pMA from Geneart. It has BioBrick prefix and suffix flanked on both sides by 70 bp of homology arms and a blasticidin resistance cassette (BsdR) for insertion of the contained BioBrick into the proper locus of pCC2FosM ([[Part:BBa_K260000|]]) by recombineering using the HincII fragment including the BioBrick. The product of this reaction is the pCC2FosMB backbone ([[Part:BBa_K260001|]]), which then contains the same BioBrick that was inside the present backbone [[Part:BBa_K260010|]] before, with BioBrick prefix and sufix maintained intact. Initially this was done with the TT_FRT_GFP BioBrick ([[Part:BBa_K260016|]]) using [[Part:BBa_K260010|]]-[[Part:BBa_K260016|]].
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This plasmid backbone is based on pMA from Geneart. It has BioBrick prefix and suffix flanked on both sides by 70 bp of homology arms and a codon-optimised blasticidin resistance cassette (BsdR) for insertion of the contained BioBrick into the proper locus of pCC2FosM ([[Part:BBa_K260000|BBa_K260000]]) by recombineering using the HincII fragment including the BioBrick. The product of this reaction is the pCC2FosMB backbone ([[Part:BBa_K260001|BBa_K260001]]), which then contains the same BioBrick that was inside the present backbone [[Part:BBa_K260010|BBa_K260010]] before, with BioBrick prefix and sufix maintained intact. Initially this was done with the TT_FRT_GFP BioBrick ([[Part:BBa_K260016|BBa_K260016]]) using [[Part:BBa_K260010|BBa_K260010]]-[[Part:BBa_K260016|BBa_K260016]].
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The pMA-@CC2FosMB-00500bp backbone is available with [[Part:BBa_K260015|]], which is composed of promoter-FRT-dhfr. If the PvuII fragment from [[Part:BBa_K260004|]]-[[Part:BBa_K260015|]] is transferred by recombineering to the the 00500 locus of [[Part:BBa_K260001|]]-[[Part:BBa_K260016|]], the intervening sequence between the FRT sites of [[Part:BBa_K260015|]] and [[Part:BBa_K260016|]] on the [[Part:BBa_K260001|]] backbone will be 500 bp exactly.
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These plasmid backbones are similar and all allow recombineering-mediated transfer of the contained BioBrick (typically [[Part:BBa_K260015|]]) to the pCC2FosMB backbone [[Part:BBa_K2600001|]]. They differ in the homology arms, which determines the locus of insertion into pCC2FosMB. The bp numbers denote final inter-FRT distance as described above:
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[[Part:BBa_K260004|]]: pMA-CC2FosMB-00500bp
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[[Part:BBa_K260005|]]: pMA-CC2FosMB-01000bp
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[[Part:BBa_K260006|]]: pMA-CC2FosMB-02000bp
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[[Part:BBa_K260007|]]: pMA-CC2FosMB-05000bp
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[[Part:BBa_K260008|]]: pMA-RQ-CC2FosMB-10000bp
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SOURCE:
 
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Geneart plasmid pMA, with 2x 70 bp from pCC2FosMB [[Part:K260001|]].
 
  
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DESIGN:
 
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The 70 bp homologies were designed to delete 100 bp of [[Part:BBa_K260001|]]-[[Part:BBa_K260016|]] and put the FRT site of [[Part:BBa_K260015|]] (if used with the present backbone [[Part:BBa_K260004|]]) at a defined distance from the FRT site of [[Part:BBa_K260016|]].
 
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 08:31, 18 October 2009

pMA-BsdR-@CC2FosM

DESCRIPTION: This plasmid backbone is based on pMA from Geneart. It has BioBrick prefix and suffix flanked on both sides by 70 bp of homology arms and a codon-optimised blasticidin resistance cassette (BsdR) for insertion of the contained BioBrick into the proper locus of pCC2FosM (BBa_K260000) by recombineering using the HincII fragment including the BioBrick. The product of this reaction is the pCC2FosMB backbone (BBa_K260001), which then contains the same BioBrick that was inside the present backbone BBa_K260010 before, with BioBrick prefix and sufix maintained intact. Initially this was done with the TT_FRT_GFP BioBrick (BBa_K260016) using BBa_K260010-BBa_K260016.


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 3076
    Illegal NheI site found at 3014
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 3082
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3076
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 3076
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 3076
    Plasmid lacks a suffix.
    Illegal XbaI site found at 3091
    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.