Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Katie Lund
Date created: 2011-05-05 11:00:00
Date modified: 2015-05-08 01:14:05
PaceBAK with Gemini
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_M36704_sequence (Version 1) |
Description
PaceBAK, the IclR associated promoter region of the aceBAK operon, has been coupled with the Gemini protein generator. PaceBAK includes numerous binding sites for IclR, which, when actively bound by pyruvate-stabilized IclR, will inhibit expression of Gemini.Notes
This part will test the efficacy of PaceBAK, the aceBAK promoter sequence to which IclR binds, in order to ensure that there isn't an issue with the promoter region itself. Though there will not be equal levels of IclR and the plasmid containing PaceBAK, we expect under high pyruvate levels there should be enough restriction to witness a decline in Gemini expression.The part will function by expressing Gemini in the absence of IclR and Pyruvate. If there is adequate amounts of pyruvate-stabilized IclR, however, the promoter region will be inhibited and Gemini will not be expressed.
The reason we had to alter the part from M36700 was because of a missing part of the promoter. In the M36707 version, the -10 region of the promoter was missing, as well as a sequence of what we believe is spacing DNA. When using multiple sources to figure out the sequence we needed, there was a mis-numbering in which the full promoter was included on the one website (biocyc.org) while the numbering on the website we actually copied the sequence from, (genome.jp) had a slightly different numbering system. Without the -10 region of the promoter, and the spacer following it, not only would the polymerase not bind, but without the geographic spacing between the binding site and the start codon, there may not be adequate space for the gene downstream to be synthesized. Thus we concluded that without the -10 region of the promoter, the polymerase will never bind and thus the downstream gene will never made even in the absence of IclR and pyruvate. As for without the spacer, even if the polymerase can bind the DNA, the downstream gene still will not be made because the ribosome does not have adequate space to bind and find the start codon.
Source
See individual parts| Access | Instance | Definition |
| public public | BBa_M36699 BBa_M36709 | BBa_M36699 BBa_M36709 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_M36699 BBa_M36709 | 1,218 219,1265 | BBa_M36699 BBa_M36709 |