Part:BBa_K4472988
Split ribozyme detecting hcat and expressing eforRED
This composite part is build out of the split ribozyme halfs, guide RNAs complementary to the stable endogenous E.coli mRNA hcat and an eforRED reporter. We assembled this part using golden gate assembly and transformed it into E.coli DH5alpha.
In our colony PCRs we were able to confirm that we have assembled the construct correctly as all our PCRs had bands at the expected height of around 1600-1700 bp.
Figure 1: Gel of Colony PCR of assembled split ribozyme - reporter constructs. Bands expected at around 1600-1700bp. Ladder: 1kb benchtop from Promega. Marked is the PCR of 2 clones with BBa_K4472990.
The promoters were too weak to get any signal in our plate reader measurements afterwards. Our model showed that stronger promoters would have had a much stronger effect, see here: https://2022.igem.wiki/uni-hamburg/model
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 331
Illegal NheI site found at 354
Illegal NheI site found at 686 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 588
Illegal XhoI site found at 135 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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