Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Cyrille Pauthenier
Date created: 2012-09-19 11:00:00
Date modified: 2015-05-08 01:13:27
Golden brick version of pSB1C3 plasmid for Golden Gate and Golden Brick cloning
| Types | DnaRegion |
| Roles | plasmid_vector Plasmid_Backbone |
| Sequences | BBa_K812050_sequence (Version 1) |
Description
This is a modified version of pSB1C3 from the registry with the J4450 negative control. The biobrick sites are modified to incorporate a BsaI restriction site that enable to the use of golden gate cloning and golden brick cloning to assemble full contructs in a single shot of digestion/ligation.The complete protocol of the goldengate can be find in the original paper: Engler C, Gruetzner R, Kandzia R, Marillonnet S (2009) Golden Gate Shuffling: A One-Pot DNA Shuffling Method Based on Type IIs Restriction
Enzymes. PLoS ONE 4(5): e5553. doi:10.1371/journal.pone.0005553
The Golden Brick assembly is an invention of the iGEM Evry 2012 Team. A complete description of the procedure can be find on our website: http://2012.igem.org/Team:Evry/GB
Notes
The negative control J4450 has been amplified with new goldenbrick extensions using PCR and re-inserted in pSB1C3 using EcoRI and PstI cloning.Source
This part is a derivation of the pSB1C3 plasmid from the registry.| Sequence Annotation | Location | Component / Role(s) |
| Golden Brick Back SpeI BsaI PstI BbsI OH5 pSB1C3 backbone E. coli His Operon terminator VR pMB1 replication origin T0 terminator Cloramphenicol resistance CDS VF2 Terminator Golden Brick Back EcoRI Bbsi OH1 BsaI XbaI | 1,27 1,6 7,12 23,27 17,22 14,17 28,2057 29,101 165,184 269,883 1050,1155 1168,1827 1941,1960 2030,2049 2058,2084 2058,2062 2063,2068 2068,2071 2073,2078 2079,2084 | feature/BioBrick engineered_region feature/misc sequence_feature sequence_feature feature/misc sequence_feature feature/misc sequence_feature feature/misc sequence_feature feature/misc sequence_feature feature/dna stem_loop feature/stem_loop primer_binding_site feature/primer_binding feature/misc sequence_feature feature/stem_loop stem_loop feature/protein CDS primer_binding_site feature/primer_binding sequence_feature feature/misc engineered_region feature/BioBrick sequence_feature feature/misc feature/misc sequence_feature feature/misc sequence_feature sequence_feature feature/misc feature/misc sequence_feature |