Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Claire Shinneman, Paul Frederick, A. Malcolm Campbell, Todd Eckdahl
Date created: 2013-01-13 12:00:00
Date modified: 2015-08-31 04:08:27
pClone Red - Device for Testing New Promoters via Golden Gate Assembly
| Types | DnaRegion |
| Roles | Reporter engineered_region |
| Sequences | BBa_J119137_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 promoter driving GFP expression with the new promoter. The new promoter will drive RFP expression in the forward direction or GFP expression if it has reverse promoter function. The part incorporate the BD18 bicistronic translational junction (see Part:BBa_J119024) engineered by Vivek Mutalik and The BIOFAB Team at biofab.org.Notes
None.Source
GGA BsmBI assembly of PCR product and BBa_J119136.| Sequence Annotation | Location | Component / Role(s) |
| GFP reverse B0030 RBS reverse Sticky end BsaI left pLacIQ1 reverse BsaI right Sticky end BD18 bicistron RBS RFP double stop | 1,720 727,738 755,758 760,765 766,836 837,842 844,847 860,944 945,1625 1620,1625 | feature/cds CDS ribosome_entry_site feature/rbs sequence_feature feature/misc sequence_feature feature/dna feature/promoter promoter feature/dna sequence_feature feature/misc sequence_feature feature/rbs ribosome_entry_site feature/cds CDS feature/stop stop_codon |