Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Claire Shinneman, Paul Frederick
Date created: 2013-01-13 12:00:00
Date modified: 2015-08-31 04:08:27
Device for Testing New Promoters via Golden Gate Assembly
| Types | DnaRegion |
| Roles | engineered_region Measurement |
| Sequences | BBa_J119138_sequence (Version 1) |
Description
This part will allow users to clone and test new promoters without gel purification or other preparation of DNA - it is ideal for teaching labs. It is a destination vector for Golden Gate Assembly (GGA) using BsaI and Ligase. A new promoter can be derived from synthetic oligos, PCR, or a plasmid clone. The new promoter must be flanked by BsaI sites that produce the 4 nt overhangs required for assembly. The left site must be 5' CGAC 3' and the right site must be 5' GCGG 3'. Successful GGA assembly replaces the reverse GFP expression casette with the new promoter. A functional new promoter will drive RFP expression in the forward direction. The part incorporate the BD18 bicistronic translational junction (see Part:BBa_J119024) engineered by Vivek Mutalik and The BIOFAB Team at biofab.org.Notes
NoneSource
GGA BsmBI assembly of PCR product from BBa_K119013 and BBa_J119136| Sequence Annotation | Location | Component / Role(s) |
| Sticky end Sticky end BsaI left BsaI right GFP reverse RBS reverse pLacIQ1 reverse RFP BD18 bicistron RBS | 23,26 843,846 28,33 849,855 34,753 760,772 781,836 933,1613 848,935 | sequence_feature feature/misc feature/misc sequence_feature feature/dna sequence_feature feature/dna sequence_feature feature/cds CDS ribosome_entry_site feature/rbs feature/promoter promoter CDS feature/cds ribosome_entry_site feature/rbs |