Difference between revisions of "Part:BBa K1985016"

 
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<partinfo>BBa_K1985016 short</partinfo>
 
<partinfo>BBa_K1985016 short</partinfo>
  
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This is an improvement on Part:BBa_K1321333. The arabinose-inducible promoter pBAD and it's transcriptional inhibitor/activator AraC was inserted into the pSB1A3 backbone.
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This part encodes Part:BBa_K1321333 in a pSB1A3 backbone. It was ligated to this backbone as it confers the ampicillin resistance that was needed for in vivo expression for our composite parts: Bba_K1985017, Bba_K1985008, Bba_K1985009, Bba_K1985013, Bba_K1985014, Bba_K1985015.
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===Validation===
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The plasmid was analysed through a diagnostic double restriction cut, using the enzymes EcoRI and PStI. This was followed by agarose gel electrophoresis. The enzymes cleave the pBAD Ara-C promoter at 1165 bp, with the remainder plasmid being 2114 bp. The sizes of the two fragments were compared with the size of the Invitrogen 1kB DNA marker and was found that our fragments were the correct.
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<!-- Add more about the biology of this part here
 
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===Usage and Biology===
 
  
 
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Revision as of 21:58, 19 October 2016


AraC-pBAD in pSB1A3

This part encodes Part:BBa_K1321333 in a pSB1A3 backbone. It was ligated to this backbone as it confers the ampicillin resistance that was needed for in vivo expression for our composite parts: Bba_K1985017, Bba_K1985008, Bba_K1985009, Bba_K1985013, Bba_K1985014, Bba_K1985015.

Validation

The plasmid was analysed through a diagnostic double restriction cut, using the enzymes EcoRI and PStI. This was followed by agarose gel electrophoresis. The enzymes cleave the pBAD Ara-C promoter at 1165 bp, with the remainder plasmid being 2114 bp. The sizes of the two fragments were compared with the size of the Invitrogen 1kB DNA marker and was found that our fragments were the correct.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961