Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Anthony Eckdahl
Date created: 2015-05-01 11:00:00
Date modified: 2015-08-31 04:08:29
rClone Red: Device for GGA Cloning and Testing RBS elements and Riboswitches
| Types | DnaRegion |
| Roles | Device engineered_region |
| Sequences | BBa_J119384_sequence (Version 1) |
Description
rClone Red allows users to clone and test new RBS elements and riboswitches without gel purification or other preparation of DNA. It is a destination vector for Golden Gate Assembly (GGA) using BsaI and ligase. A new RBS 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 RBS or riboswitch. Transcription will be initiated in the direction of the RFP coding sequence.Notes
None.Source
BsmBI GGA mutagenesis of J119361.| Sequence Annotation | Location | Component / Role(s) |
| P5 promoter BsaI left GFP reverse B0034 RBS reverse P2 promoter reverse BsaI right RFP | 1,36 42,47 55,774 781,792 803,848 849,854 862,1542 | promoter feature/promoter feature/dna sequence_feature feature/cds CDS ribosome_entry_site feature/rbs feature/promoter promoter feature/dna sequence_feature CDS feature/cds |