Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Claire Shinneman, Paul Frederick
Date created: 2013-01-10 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 Reporter |
| Sequences | BBa_J119136_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 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. The left site must be 5' CGAC 3' and the right site must be 5' GCGG 3'. Successful GGA assembly replaces the double terminator in with the new promoter. Upon assembly, a functional new promoter with activity to the right should cause RFP expression. A promoter with activity to the left will cause GFP expression. 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
PCR product from amplification of E5500 ws cloned into BBa_J119127.| Access | Instance | Definition |
| public public | BBa_J119135 BBa_J119127 | BBa_J119135 BBa_J119127 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_J119135 BBa_J119127 | 1,741 750,1632 | BBa_J119135 BBa_J119127 |