Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Anthony Eckdahl
Date created: 2015-06-28 11:00:00
Date modified: 2015-08-31 04:08:29
rClone Blue: Device for GGA Cloning and Testing RBS elements and Riboswitches
| Types | DnaRegion |
| Roles | engineered_region Device |
| Sequences | BBa_J119389_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. The level of expression of RFP will depend on the efficiency of the newly cloned RBS or riboswitch.Notes
None.Source
PCR of J119139 and J119384 and BbsI GGA.| Sequence Annotation | Location | Component / Role(s) |
| B0034 RBS reverse GFP reverse BsaI left P5 promoter BsaI right Amil CP Blue P2 promoter reverse | 781,792 55,774 42,47 1,36 849,854 862,1530 803,848 | feature/rbs ribosome_entry_site CDS feature/cds sequence_feature feature/dna feature/promoter promoter sequence_feature feature/dna feature/cds CDS promoter feature/promoter |