Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Todd Eckdahl
Date created: 2012-10-29 12:00:00
Date modified: 2015-08-31 04:08:27
For Testing New Promoters via Golden Gate Assembly with BsmBI
| Types | DnaRegion |
| Roles | engineered_region Measurement |
| Sequences | BBa_J119127_sequence (Version 1) |
Description
This part allows users to clone and test new promoters without gel purification or other preparation of DNA. It is a destination vector for Golden Gate Assembly (GGA) using BsmBI and Ligase. A new promoter can be derived from synthetic oligos, PCR, or a plasmid clone. The new promoter must be flanked by BsmBI sites that produce the 4 nt overhangs required for assembly (see [[Part:BBa_J119022]] as an example, or the picture below). Successful GGA assembly replaces the double terminator in BBa_J119127 with the new promoter. Upon assembly, a functional new promoter should cause RFP expression. BBa_J119127 incorporates the BD18 bicistronic translational junction (see [[Part:BBa_J119024]]) engineered by Vivek Mutalik and The BIOFAB Team at biofab.org.See the full GGA protocol online.
[[Image:J100028.png]]
Notes
None.Source
Golden Gate assembled of PCR product and BBa_J100091 using BsaI and Ligase.| Sequence Annotation | Location | Component / Role(s) |
| BsmBI sticky end left BsmBI cuts left TT B0014 BsmBI cuts right BsmBI sticky end right BD18 biscistron leader RBS RBS for RFP Stop for BD18 leader RFP E1010 Start for RFP Double stop for E1010 | 1,4 6,11 12,106 107,112 114,117 118,205 135,143 188,196 201,203 203,883 203,205 878,883 | sequence_feature feature/dna feature/dna sequence_feature stem_loop feature/stem_loop sequence_feature feature/misc feature/dna sequence_feature feature/misc sequence_feature feature/rbs ribosome_entry_site feature/rbs ribosome_entry_site feature/stop stop_codon feature/protein CDS start_codon feature/start stop_codon feature/stop |