Part:BBa_J119022

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 [http://gcat.davidson.edu/mediawiki-1.15.0/index.php/Golden_Gate GGA.] You can follow a [http://gcat.davidson.edu/mediawiki-1.15.0/index.php/Golden_Gate_Assembly_protocol 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 [http://gcat.davidson.edu/RFP/ 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