Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Sigrun Stulz
Date created: 2016-10-13 11:00:00
Date modified: 2016-10-14 01:15:18
Vector containing Hisx6 and Sortase tag LPETGG
| Types | DnaRegion |
| Roles | Plasmid_Backbone plasmid_vector |
| Sequences | BBa_K2144101_sequence (Version 1) |
Description
This vector is constructed to simplify the usage of the transpeptidase Sortase A for future iGEM teams. Following a BamHI restriction site we constructed a synthetic sequence consisting of three main elements:1) A linker (GSSS) to allow for limited mobility of the attached peptides and facilitate correct folding
2) A Sortase A tag (LPETGG) that is used to conjugate the proteins to poly-glycines
3) A standard Hisx6 tag commonly used for Immobilized Metal Affinity Chromatography (IMAC)
This sequence follows a BamHI restriction site which can be used by teams to clone in whichever peptide they would like to conjugate using Sortase A.
Notes
BamHI is a very convenient restriction enzyme for our application as its restriction site will code for a glycine and a serine, both of which are often used in linkers and tend to be unproblematic in such sequences.Source
The plasmid was constructed using the iGEM-supplied pSB1C3 and a synthetically produced sequence.| Sequence Annotation | Location | Component / Role(s) |
| BBa_R0010 CAP -35 -10 LacI start BBa_B0034 conserved ori barcode BBa_B0010 stem_loop BBa_B0012 T7 TE polya stop CmR Hisx6 Sortase A Tag Stop | 1,200 89,126 137,142 161,166 166,200 173,173 209,220 213,216 254,842 908,932 941,1020 952,995 1029,1069 1036,1055 1056,1069 1062,1062 1140,1799 2077,2094 2059,2076 2095,2097 | engineered_region feature/BioBrick non_covalent_binding_site feature/binding promoter feature/promoter promoter feature/promoter non_covalent_binding_site feature/binding start_codon feature/start feature/BioBrick engineered_region feature/conserved conserved_region sequence_feature feature/misc engineered_tag feature/barcode engineered_region feature/BioBrick stem_loop feature/stem_loop engineered_region feature/BioBrick feature/stem_loop stem_loop feature/polya polyA_site feature/stop stop_codon CDS feature/cds CDS feature/cds feature/cds CDS feature/stop stop_codon |