Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Todd Eckdahl and A. Malcolm Campbell
Date created: 2012-01-06 12:00:00
Date modified: 2015-08-31 04:08:26
RFP GGA destination vector BsaI
| Types | DnaRegion |
| Roles | engineered_region Measurement |
| Sequences | BBa_J119044_sequence (Version 1) |
Description
The construct allows for the cloning and testing of new promoter sequences. It is a destination vector for Golden Gate Assembly using BsaI and Ligase, based on part J100028. 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 (see J119022). Assembly replaces the double terminator in the destination vector with the new promoter. Upon assembly, a functional new promoter will be expected to cause RFP expression. The destination vector also incorporates the BD18 bicistronic translational junction (see J119024) engineered by Vivek Mutalik and The BIOFAB Team at biofab.org.Notes
Golden Gate Assembly, replacement of TT with promoter, BD18 bicistronic translational junction engineered by Vivek Mutalik and The BIOFAB Team at biofab.org.Source
De novo DNA synthesis.| Sequence Annotation | Location | Component / Role(s) |
| Bba prefix BsaI sticky end left BsaI cuts left TT B0014 Bsa cuts right BsaI sticky end right BD18 bicistron leader RBS RFP RBS Stop for BD18 Start RFP RFP optimized for E. coli Double Stop BBa suffix | 1,22 23,26 28,33 34,128 129,134 136,139 140,227 157,165 210,218 223,225 225,227 225,905 900,905 906,926 | engineered_region feature/BioBrick feature/dna sequence_feature sequence_feature feature/misc stem_loop feature/stem_loop feature/misc sequence_feature feature/dna sequence_feature feature/misc sequence_feature ribosome_entry_site feature/rbs feature/rbs ribosome_entry_site stop_codon feature/stop start_codon feature/start feature/protein CDS stop_codon feature/stop feature/BioBrick engineered_region |