Basic Vector Information
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.
Basic Vector Information | |||
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Vector Name | pcDNA3.4 | Length | 6011 bp |
Type | Mammalian Expression Vectors | Source | Thermo Fisher Scientific |
Promoter | CMV |
pcDNA3.4 vector Vector Map
pcDNA3.4 vector Sequence
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 JOURNAL Exported Saturday, November 14, 2020 from SnapGene 3.2.1 http://www.snapgene.com 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"
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