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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