Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Karmella Haynes
Date created: 2013-04-18 11:00:00
Date modified: 2015-08-31 04:08:35
Kozak-XbaI-NLS-6His-stop
| Types | DnaRegion |
| Roles | plasmid_vector Plasmid_Backbone |
| Sequences | BBa_J176171_sequence (Version 1) |
Description
Mammalian expression vector (without a promoter) designed to add a eukaryotic RBS (Kozak) upstream (in-frame) and a nuclear localization tag, 6-histidine tag, and stop codon downstream (in-frame) of a protein coding insert. This method only works for Silver Standard (RFC 23) inserts.Notes
- A Kozak-XbaI-NLS-6His-stop with SpeI sticky overhangs was inserted into the XbaI site.
- Sequencing data confirms proper orientation of the inserted part.
- XbaI site can accept any Silver Standard (RFC 23) protein coding region that is cut with XbaI/SpeI.
- Single-cut vectors can easily re-ligate: It is highly recommended that the user dephosphorylates the XbaI-cut vector prior to ligation.
- Insertion of a XbaI/SpeI insert is non-directional: It is high recommended that the user selects at least 3 colonies after ligation check (vi sequencing or some other method) for proper orientation of the insert.
Source
Built from BBa_J176122| Sequence Annotation | Location | Component / Role(s) |
| DD122 B pCMV remnant MCS: SpeI BstXI EcoRI PstI EcoRV BstXI NotI XhoI Kozak XbaI NLS 6-His A - Stop - S - stop BGH reverse DD123 B BGH polyA f1 origin SV40 promoter/ origin Puromycin R SV40 polyA Ampicillin R bla promoter | 186,203 232,249 250,304 311,328 329,334 335,355 356,373 374,386 418,435 426,444 424,648 694,1122 1127,1471 1475,2074 2288,2417 3615,4475 4476,4574 | primer_binding_site feature/primer_binding sequence_feature feature/misc feature/misc sequence_feature feature/rbs ribosome_entry_site feature/misc sequence_feature feature/tag tag tag feature/tag feature/stop stop_codon feature/primer_binding primer_binding_site feature/primer_binding primer_binding_site polyA_site feature/polya sequence_feature feature/misc feature/promoter promoter feature/protein CDS feature/polya polyA_site feature/protein CDS promoter feature/promoter |