DNA

Part:BBa_J119022

Designed by: Caleb J. Carr   Group: Eckdahl_Lab   (2011-10-19)

HPP Bsa J23100 promoter in pSB1A2

J119022 can be used with either Part:BBa_J119044 or Part:BBa_J100091 for Golden Gate Assembly (GGA) to move promoter Part:BBa_J23100 upstream of RFP where the transcriptional terminator (TT) used to be located. J119022 is an ideal way to start learning how easy it is to swap two parts using GGA. You can follow a complete protocol for use in teaching labs.

J119022 contains a BsaI-flanked Part:BBa_J23100 promoter in pSB1A8 (Part:BBa_J119043) for BsaI/Ligase insertion into Part:BBa_J119044 or Part:BBa_J100091.

The Part:BBa_J23100 promoter is designed for simultaneous BsaI digestion and ligation into Part:BBa_J119044 or Part:BBa_J100091 as a positive control for use in Golden Gate Assembly. J119022 is the first promoter to be use as part of the Registry of Functional Promoters.

J119022 is composed of: a BsaI site on the left cutting to the right; a sticky end CGAC; Part:BBa_J23100 promoter; a different sticky end of GCGG; and BsaI site which cuts to the left. The fragment is 57bp long after digestion. It should then be ligated into (Part:BBa_J119044) or Part:BBa_J100091.


Sequence and Features

Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 15
    Illegal SpeI site found at 80
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 39
    Illegal NheI site found at 62
    Illegal SpeI site found at 80
    Illegal NotI site found at 6
    Illegal NotI site found at 87
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 15
    Illegal SpeI site found at 80
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 15
    Illegal SpeI site found at 80
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 22
    Illegal BsaI.rc site found at 73
    Illegal XbaI site found at 15
    Illegal SpeI site found at 80


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