pCDNA3.0 Plasmid

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  • Model: PVT1001
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pCDNA3.0

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pCDNA3.0,Plasmid pCDNA3.0,pCDNA3.0 vector

 

 

pCDNA3.0 Information

Promoter: CMV promoter

Replicon: ColE1 ori, F1 ori

Terminator: bGH poly (A) signal

Plasmid classification: lactation serial plasmid; lactation expression plasmid; pCDNA series plasmid.

Plasmid size: 5446bp

Plasmid label: None

Prokaryotic resistance: ampicillin Ampicillin (100 g/ml)

Screening marker: neomycin Neomycin /G418

Cloning strains: E. coli DH5 and E.

Culture conditions: 37 C, aerobic, LB

Expression host: mammalian cells such as 293T

Culture conditions: 37 C, 5%CO2

Induction mode: no need to induce

5'sequencing primers: pCDNA3.1-F (CTAGAGAACCCACTGCTTAC)

3'sequencing primers: pCDNA3.1-R (TAGAAGGCACAGTCGAGG)

Remarks: commonly used mammalian cell eukaryotic expression vectors

 

pCDNA3.0 Description

PCDNA 3.0 vector is specially designed for high level stable and transient expression in mammalian cells. Human cytomegalovirus early promoter (CMV) can be highly expressed in a wide range of mammalian cells. In addition, the neomycin resistance gene was provided for screening stable cell lines, which could be screened by G418.

 

 pcDNA3.0 is 5.4 kb vectors  designed for high-level stable and transient expression in mammalian hosts. High-level stable and non-replicative transient expression can be carried out in most mammalian cells. The vectors contain the following elements: Human cytomegalovirus immediate-early (CMV) promoter for high-level expression in a wide range of mammalian cells.  Neomycin resistance gene for selection of stable cell lines.
        pcDNA3.0 vector is designed for high-level, constitutive expression in a variety of mammalian cell lines. It contains a Geneticin selectable marker and a forward-orientation multiple cloning site. High-level stable and non-replicative transient expression can be carried out in most mammalian cells. The vectors contain the following elements: Human cytomegalovirus immediate-early (CMV) promoter for high-level expression in a wide range of mammalian cells Multiple cloning sites orientations to facilitate cloning .Neomycin resistance gene for selection of stable cell lines Episomal replication in cells lines that are latently infected with SV40 or that express the SV40 large T antigen.

 

                        CMV promoter: bases 209-863                                    T7 promoter: bases 864-882
                        Polylinker: bases 889-994                                            Sp6 promoter: bases 999-1016
                        BGH poly A: bases 1018-1249                                     SV40 promoter: bases 1790-2115
                        SV40 origin of replication: bases 1984-2069                Neomycin ORF: bases 2151-2945
                        SV40 poly A: bases 3000-3372                                    ColE1 origin: bases 3632-4305
                        Ampicillin ORF: bases 4450-5310
 
 
pCDNA3.0 Multiple cloning site
pCDNA3.0-vector
 
 
 
pCDNA3.0 Sequence
 
LOCUS       Exported                5446 bp ds-DNA    circular SYN 29-10-2015
KEYWORDS    Untitled
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5446)
  AUTHORS   admin
  TITLE     Direct Submission
  JOURNAL   Exported 2015-10-29  
FEATURES             Location/Qualifiers
     source          1..5446
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(998..1016)
                     /note="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    1042..1266
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1312..1740
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1754..2084
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      1935..2070
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             2151..2945
                     /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    3119..3240
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3289..3305)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    3313..3329
                     /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(3337..3367)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3382..3403
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3691..4279)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4450..5310)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
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