Part:BBa_K3424025
pPhoA-insulator-GFP
In the absence of a lab, The University of Florida 2020 iGEM team offers a contribution to further understand and improve part BBa_K1139201. To increase the expression of Part BBa_K1139201 designed by iGem Tokyo 2013 Team, the RiboJ insulator sequence can be added between the pPhoA promoter and the downstream sequences. Insulators protect against unexpected interactions between neighboring sequences in a genetic circuit (Clifton 2). A common insulator, RiboJ, is made up of the sTRSV-ribozyme, along with a subsequent 23-nucleotide hairpin sequence (Luo 3). The hairpin structure helps expose the ribosome binding site, so that translation can be increased for the downstream sequence transcripts (Luo 3). Therefore, insulators, and specifically, the RiboJ sequence, can be used to increase the efficacy of gene and protein expression in future experimental genetic constructs. More information on literature supporting this improvement can be found on our wiki: our contribution for UFlorida 2020 wiki. By including the RiboJ sequence between the pPhoA and the subsequent genes in Part BBa_K1139201, the expression of sfGFP protein will be increased in future experiments that quantify absolute fluorescence as the output of an experimental design.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 837
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