Basic Vector Information
EXP4(+) 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 ℃.
EXP4(+) vector Sequence
LOCUS Exported 6724 bp ds-DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector EXP4(+), complete sequence. ACCESSION EF028663 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6724) AUTHORS Meima ME, Weening KE, Schaap P. TITLE Vectors for expression of proteins with single or combinatorial fluorescent protein and tandem affinity purification tags in Dictyostelium JOURNAL Protein Expr. Purif. 53 (2), 283-288 (2007) PUBMED 17296313 REFERENCE 2 (bases 1 to 6724) AUTHORS Meima ME, Weening KE, Schaap P. TITLE Direct Submission JOURNAL Submitted (27-SEP-2006) School of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, United Kingdom REFERENCE 3 (bases 1 to 6724) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..6724 /organism="Cloning vector EXP4(+)" /lab_host="Dictyostelium discoideum" /mol_type="other DNA" /note="Cloning vector Exp4(+) was constructed by Dynes et al., (Genes & Development 8, 948-958, 1994) from pATSP (Dynes et al., Proc.Natl.Acad.Sci 86, 7966-7970, 1989), a derivative of the commercial vector pAT153" /db_xref="taxon:409536" misc_feature 1..30 /label=multipe cloning site /note="multipe cloning site" regulatory 42..1022 /regulatory_class="terminator" /note="2H3 terminator" CDS 1755..2585 /codon_start=1 /product="neomycin phosphotransferase" /label=neomycin phosphotransferase /protein_id="ABK40467.1" /translation="MDGEDVQAGSFRMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSD AAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWL LLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAG LVDQDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCG RLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" CDS 1791..2585 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" promoter 3475..3579 /gene="bla" /label=AmpR promoter CDS 3580..4440 /codon_start=1 /product="beta-lactamase" /label=beta-lactamase /protein_id="ABK40466.1" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 4611..5199 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" regulatory 6416..6724 /regulatory_class="promoter" /note="actin15 promoter"
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