Part:BBa_J100091:Design
For Testing New Promoters via Golden Gate Assembly
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 906 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1
Illegal SpeI site found at 907
Illegal PstI site found at 921
Illegal NotI site found at 7
Illegal NotI site found at 914 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1
- 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 907 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 1
Illegal XbaI site found at 16
Illegal SpeI site found at 907
Illegal PstI site found at 921
Illegal AgeI site found at 779
Illegal AgeI site found at 891 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 129
Illegal BsaI.rc site found at 28
Design Notes
The major question is what plasmid to use. Since both Part:BBa_J119044 and Part:BBa_J100091 both contain a transcriptional terminator (TT), this may lead to recombination if more than two TTs are in the same plasmid. We have some evidence that Part:BBa_J119044 can undergo some recombination when cloned into the plasmid pSB1A8 Part:BBa_J119043. Therefore, we recommend Part:BBa_J119044 and Part:BBa_J100091 be cloned into a modified version of pSB1A2 that has the Bsa I site removed from the ampicillin gene. We call this modified plasmid pSB1A2 BR for Bsa I Removed.
Source
This part was engineered by Todd Eckdahl and Malcolm Campbell. They devised this as part of a national effort to have undergraduates contribute to a registry of functional promoters (RFP; [http://gcat.davidson.edu/RFP/ Registry of Functional Promoters]). There is a very similar part called J119044 Part:BBa_J119044. The only difference between these two parts is that J00091 uses E1010 Part:BBa_E1010 version of RFP while Part:BBa_J119044 uses an E. coli-optimized set of codons and was produced by GeneArt.