Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Claire Shinneman
Date created: 2013-01-22 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_J119139_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 ampil (Blue) gene expression casette 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
BsmBI GGA of J119136 with PCR product from J100103.| Sequence Annotation | Location | Component / Role(s) |
| GFP reverse RBS reverse Sticky end Sticky end BD18 bicistron RBS RFP BD18 bicistron RBS ampil CP (blue) P2 (strong) BsaI right BsaI left | 1,720 727,741 772,775 1590,1593 1594,1678 1679,2359 1452,1536 783,1451 1536,1582 1583,1588 777,782 | feature/cds CDS ribosome_entry_site feature/rbs sequence_feature feature/misc sequence_feature feature/misc ribosome_entry_site feature/rbs CDS feature/cds feature/rbs ribosome_entry_site CDS feature/cds promoter feature/promoter sequence_feature feature/misc sequence_feature feature/dna |