Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Cyrille Pauthenier
Date created: 2012-09-23 11:00:00
Date modified: 2015-05-08 01:13:27
Golden brick version of pSB1K3 plasmid for Golden Gate and Golden Brick cloning
| Types | DnaRegion |
| Roles | Plasmid_Backbone plasmid_vector |
| Sequences | BBa_K812051_sequence (Version 1) |
Description
This is a modified version of pSB1K3 from the registry with the J4450 negative control. The biobrick sites are modified to incorporate a BsaI restriction site that enables 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 pSB1K3 plasmid from the registry.| Sequence Annotation | Location | Component / Role(s) |
| Goldenbrick suffix SpeI BsaI OH5 BbsI PstI E. coli his Operon terminator VR pMB1 rep origin Kanmycin resistance marker CDS Repeat region VF2 Terminator Goldenbrick prefix EcoRI BbsI OH1 BsaI XbaI | 1,27 1,6 7,12 14,17 17,22 23,27 29,100 164,183 268,882 1104,1919 2024,2067 2074,2093 2163,2182 2191,2217 2191,2195 2196,2201 2201,2204 2206,2211 2212,2217 | feature/BioBrick engineered_region feature/misc sequence_feature sequence_feature feature/misc sequence_feature feature/misc sequence_feature feature/misc sequence_feature feature/misc stem_loop feature/stem_loop primer_binding_site feature/primer_binding feature/misc sequence_feature feature/misc sequence_feature sequence_feature feature/misc feature/primer_binding primer_binding_site stem_loop feature/stem_loop engineered_region feature/BioBrick sequence_feature feature/misc sequence_feature feature/misc feature/misc sequence_feature feature/misc sequence_feature feature/misc sequence_feature |