Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Bernardo Pollak y Simon Alamos
Date created: 2012-09-21 11:00:00
Date modified: 2015-05-08 01:13:09
psb1C3_IntS recombination plasmid with RFP
| Types | DnaRegion |
| Roles | Composite engineered_region |
| Sequences | BBa_K743010_sequence (Version 1) |
Description
psb1C3_IntS recombination plasmid is designed to integrate any desired biobricks or other sequences into Synechocystis PCC6803 chromosome and/or pSYSX endogenous plasmid while interrupting CopS gene coding sequence.Synechocystis naturally undergoes double homologous recombination so the use of recombination sites flanking prefix and suffix were designed with complete homology to CopS gene, present in the chromosome and in a endogenous plasmid.
As CopS is essential for copper stress tolerance in this bacteria, by using this plasmid a succeptible strain incapable of thriving in the enviroment is created.(see references for more information)
Notes
The recombination sites are just 450bp long so that complete homology with any version of the CopS gene is achieved (i.e chromosomic and plasmidial versions)Though according to literature 1300 bp is the optimum size for recombination sites in Synechocystis, 450 bp yields reasonably high transformation efficiency.
Source
Recombination sites were PCR amplified from Synechocystis PCC6803 DNA. The rest of the plasmid comes from psb1C3 sequence.| Sequence Annotation | Location | Component / Role(s) |
| Spectinomycin resistance promoter Spectinomycin resistance Verification forward primer binding site B0055 terminator three stop codons TOPO A cloning site BBa_B0043 lacI promoter BBa_E1010 mRFP stop codons B0015 double terminator biobrick Suffix Biobrick Prefix TOPO A cloning site three stop codons B0054 terminator Verification revers primer binding site 5?? recombination site 3' recombination site plasmid backbone | 1,219 220,1011 1012,1031 1032,1110 1111,1121 1112,1156 1157,1356 1383,2057 2058,2063 2097,2225 2226,2246 1135,1156 2247,2259 2260,2271 2272,2340 2341,2360 2366,2832 4589,5032 2833,4588 | promoter feature/promoter CDS feature/protein primer_binding_site feature/primer_binding feature/stem_loop stem_loop feature/stop stop_codon feature/conserved conserved_region promoter feature/promoter CDS feature/protein stop_codon feature/stop stem_loop feature/stem_loop feature/conserved conserved_region conserved_region feature/conserved feature/conserved conserved_region feature/stop stop_codon feature/stem_loop stem_loop feature/primer_binding primer_binding_site sequence_feature feature/dna sequence_feature feature/dna conserved_region feature/conserved |