Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Mike Quaney, Todd Eckdahl
Date created: 2014-07-09 11:00:00
Date modified: 2015-08-31 04:08:28
tClone Red: Device for Testing Transciptional Terminators and Riboswitches via Golden Gate Assembly
| Types | DnaRegion |
| Roles | Device engineered_region |
| Sequences | BBa_J119361_sequence (Version 1) |
Description
This part (also called tClone Red) will allow users to clone and test new transcriptional terminators and riboswitches that function by antitermination without gel purification or other preparation of DNA. It is a destination vector for Golden Gate Assembly (GGA) using BsaI and ligase. A new terminator or riboswitch can be derived from synthetic oligos, PCR, or a plasmid clone. For proper assembly, the new insert must have the appropriate 4 nt sticky ends or be flanked by BsaI sites that produce the sticky ends. With reference to the top strand, the left site must be 5' CGAC 3' and the right site must be 5' GCGG 3'. BsaI always produces a 5' overhang sticky end. Successful GGA assembly replaces the reverse promoter driving GFP expression with the new terminator or riboswitch. Transcription will be initiated in the direction of the RFP coding sequence. Whether or not transcription proceeds to the RFP gene is determined by the strength of the terminator or whether termination or antitermination occurs at the riboswitch. 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
Part J119137 was used to clone the pTac promoter. The resulting plasmid was used in BsmBI GGA to produce J119361.| Sequence Annotation | Location | Component / Role(s) |
| P5 promoter BsaI left GFP reverse B0030 RBS reverse P2 promoter reverse BsaI right BD18 C dog RBS RFP | 1,36 42,47 55,774 781,795 803,848 849,854 860,947 945,1625 | promoter feature/promoter feature/dna sequence_feature CDS feature/cds ribosome_entry_site feature/rbs feature/promoter promoter sequence_feature feature/dna feature/rbs ribosome_entry_site CDS feature/cds |