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 in pSB1A8 Part:BBa_J119043 with a BsaI site on the left cutting to the right, with sticky end CGAC being freed up, followed by Part:BBa_J23100 promoter, followed up by a different sticky end of GCGG which is freed up by the following 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
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 15
Illegal SpeI site found at 80 - 12INCOMPATIBLE 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 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 15
Illegal SpeI site found at 80 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 15
Illegal SpeI site found at 80 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 22
Illegal BsaI.rc site found at 73
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