The pcDNA3.4 circular vector is a constitutive mammalian expression vector designed to deliver exceptionally high levels of transgene expression. It can be used in suspension-adapted cells, such as Expi293F™, FreeStyle 293-F, and FreeStyle CHO-S cells, for transient protein expression. The vector can also be used as a Geneticin®-selectable expression plasmid for engineering stable cell lines. The enhanced, full-length CMV promoter and other expression elements typically enable higher expression levels than pcDNA 3.1-, 3.2-, and 3.3-based expression constructs.
| 载体基本信息 | |||
| 载体名称 | pcDNA3.4 | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 6011 bp | 载体类型 | Mammalian Expression Vectors |
| 载体来源 | Thermo Fisher Scientific | 启动子 | CMV |
pcDNA3.4 载体载体图谱
pcDNA3.4 载体载体序列
LOCUS Exported 6011 bp ds-DNA circular SYN 08-JAN-2019
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCDNA3.4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6011)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6011
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 50..429
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 430..633
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 787..1375
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(1258..1269)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
polyA_signal 1441..1489
/note="HSV TK poly(A) signal"
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 1691..2119
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2133..2462
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2313..2448
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 2529..3323
/codon_start=1
/gene="aph(3)-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3499..3620
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(3669..3685)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3693..3709
/bound_moiety="lac repressor encoded by lacI"
/note="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(3717..3747)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3762..3783
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4071..4659)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4830..5690)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5691..5795)
/gene="bla"
/note="AmpR promoter"
