Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Anthony Eckdahl
Date created: 2015-03-21 12:00:00
Date modified: 2015-08-31 04:08:29
Psimp1 Promoter with Four Consensus Variants
| Types | DnaRegion |
| Roles | Regulatory promoter |
| Sequences | BBa_J119376_sequence (Version 1) |
Description
Mutations were introduced into the Ptac promoter using NNNNNN in place of the TTGACA of the -35 region. Synthetic oligonucleotides carry the 4096 possible sequences were cloned via Golden Gate Assembly into pClone Red. A total of 81 different promoters were picked. The DNA sequence of the 81 mutant promoters was determined and their strength was measured by fluorometry (see BBa_J119375). A weighted method of consensus building was developed that assigned a score to each base in each -35 region based on the RFP expression of the mutant promoter that contained it. The resulting pClone -35 Consensus produced is shown below.Four variants of the consensus were cloned into a simplified promoter sequence to test the effect of sequence context surrounding the -35 region. The simplified promoter is called Psimp1 and is derived from De Mey et al. (2007) MC Biotechnol 7:34. Below are images and fluorescent data that show the strength of the Psimp1 promoter with the TTGACA consensus and four variants of the pClone -35 Consensus sequence.
Notes
None.Source
Synthetic oligonucleotides.| Sequence Annotation | Location | Component / Role(s) |
| -35 region -10 region pClone -35 variants | 20,25 43,48 20,25 | promoter feature/promoter feature/promoter promoter sequence_alteration feature/mutation |