Basic Vector Information
pTS1012 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pTS1012 vector Sequence
LOCUS 40924_44369 5778 bp DNA circular SYN 18-DEC-2018 DEFINITION Expression vector pTS1012, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5778) AUTHORS Scheller L, Strittmatter T, Fuchs D, Bojar D, Fussenegger M. TITLE Generalized extracellular molecule sensor platform for programming cellular behavior JOURNAL Nat. Chem. Biol. 14 (7), 723-729 (2018) PUBMED 29686358 REFERENCE 2 (bases 1 to 5778) AUTHORS Scheller L, Strittmatter T. TITLE Direct Submission JOURNAL Submitted (06-NOV-2017) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 4058, Switzerland REFERENCE 3 (bases 1 to 5778) TITLE Direct Submission REFERENCE 4 (bases 1 to 5778) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem. Biol."; date: "2018"; volume: "14"; issue: "7"; pages: "723-729" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (06-NOV-2017) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 4058, Switzerland" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..5778 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 21..400 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 401..604 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" promoter 649..667 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" sig_peptide 708..767 /label=Ig-kappa leader /note="leader sequence from mouse immunoglobulin kappa light chain" CDS 786..1298 /codon_start=1 /label=Nluc /note="NanoLuc(R) luciferase" /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW RLCERILA" polyA_signal 1359..1583 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1629..2057 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2071..2400 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 2467..3258 /codon_start=1 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" polyA_signal 3435..3568 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3605..3621) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(3629..3645) /label=lac operator /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(3653..3683) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(3698..3719) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4007..4592) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4766..5623) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(5624..5728) /label=AmpR promoter
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