Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Samantha Huckuntod
Date created: 2011-12-14 12:00:00
Date modified: 2015-08-31 04:08:26
RFP GGA BD18 destination vector - BbsI and BsaI
| Types | DnaRegion |
| Roles | Measurement engineered_region |
| Sequences | BBa_J119023_sequence (Version 1) |
Description
The construct allows for the cloning and testing of new promoter sequences. It is a destination vector for Golden Gate Assembly 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 (see J119022). Assembly replaces the double terminator in the destination vector with the new promoter. Upon assembly, a functional new promoter will be expected to cause RFP expression. The destination vector also incorporates the BD18 bicistronic translational junction engineered by Vivek Mutalik and The BIOFAB Team at biofab.org.Notes
Golden Gate Assembly, replacement of TT with promoter, BD18 bicistronic translational junction engineered by Vivek Mutalik and The BIOFAB Team at biofab.org.Source
Synthetic oligos cloned into existing part.| Sequence Annotation | Location | Component / Role(s) |
| BBa_prefix BsaI cuts left TT B0014 BsaI cuts right BsaI Word 1 BsaI Word 2 BD18 bicistron stop for BD18 Start GOI RFP E1010 with 4 pt mutations Stop E1010 BBa_suffix BbsI cuts left BbsI Word 1 BbsI cuts right BbsI Word 2 C--> T for E. coli C--> T for E. coli C--> T for E. coli C--> T for E. coli leader RBS GOI RBS | 1,22 34,39 40,134 135,140 29,32 142,145 158,245 241,243 243,245 243,923 918,923 924,944 23,28 17,20 146,151 154,157 260,260 545,545 704,704 863,863 175,183 228,236 | engineered_region feature/BioBrick feature/misc sequence_feature feature/stem_loop stem_loop sequence_feature feature/misc sequence_feature feature/dna sequence_feature feature/dna CDS feature/cds stop_codon feature/stop start_codon feature/start feature/protein CDS stop_codon feature/stop feature/BioBrick engineered_region feature/misc sequence_feature feature/dna sequence_feature feature/misc sequence_feature feature/dna sequence_feature feature/s_mutation point_mutation feature/s_mutation point_mutation feature/s_mutation point_mutation point_mutation feature/s_mutation feature/rbs ribosome_entry_site feature/rbs ribosome_entry_site |