Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Phoebe Parrish
Date created: 2013-07-12 11:00:00
Date modified: 2015-08-31 04:08:23
eCDM8 and GFP Riboswitch into pSB1A8
| Types | DnaRegion |
| Roles | sequence_feature DNA |
| Sequences | BBa_J100114_sequence (Version 1) |
Description
High promoter/RBS combination with eCDM8 (part J119303) plus promoter/riboswitch/GFP construct (J100079), ligated into pSB1A8 (J119043). This part was constructed using iPCR and PCR to amplify the desired sequences and add BsaI sites, and Golden Gate Assembly to ligate the sequences together. eCDM8 demethylates caffeine to produce theophylline, which in turn binds to the riboswitch, allowing the translation of GFP mRNA, producing green fluorescent protein as a reporter of theophylline production.Notes
This construct was built using iPCR primers to amplify the promoter/RBS/eCDM8 construct (J119303) on pSB1A8 and add to BsaI sites to the end of the amplified sequence. This amplification removed the stuffer and the SpeI site from pSB1A8. PCR primers were used to amplify the promoter/riboswitch/GFP construct (J100079) and to add BsaI sites to the end of this sequence. GGA was then used to ligate these two amplified parts together.Source
All parts taken from the registry.| Sequence Annotation | Location | Component / Role(s) |
| P5 Promoter BD2 Bicistronic Translational Junction eCDM8 Stop T5 Promoter Superfolder GFP Riboswitch | 1,36 37,124 122,3271 3269,3271 3272,3322 3429,4150 3373,3430 | promoter feature/promoter feature/rbs ribosome_entry_site feature/dna sequence_feature feature/stop stop_codon feature/promoter promoter feature/dna sequence_feature sequence_feature feature/misc |