Regulatory

Part:BBa_K1985016

Designed by: Rita Adriani   Group: iGEM16_Kent   (2016-10-14)

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 to express our composite parts: BBa_K1985017, BBa_K1985008, BBa_K1985009, BBa_K1985013, BBa_K1985014 and BBa_K1985015.

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


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.

Figure 1.1% agarose gel of the restriction digest of BBa_K1985016 in pSB1A3 plasmid backbone with EcoRI and PstI



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