pCDNA3.1 (- )/ Myc- His B/ pCDNA3.1 (- )/ Myc- His B质粒/ pCDNA3.1 (- )/ Myc- His B Plasmid

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pCDNA3.1(-)/Myc-His B Information

Promoter: CMV
Replicator: pUC ori, F1 ori
Terminator: BGH poly (A) signal
Plasmid classification: mammalian cell carrier; protein overexpression plasmid
Plasmid size: 5520bp
Plasmid label: 6X His, myc
Prokaryotic resistance: ampicillin Amp
Screening markers: neomycin Neo
Cloned strain: DH5 alpha
Culture conditions: 37 centigrade, aerobic, LB
Expression host: mammalian cells

pCDNA3.1(-)/Myc-His B Description

pcDNA3.1/myc-His(-) A, B, and C are 5.5 kb vectors designed for overproduction of recombinant proteins in mammalian cell lines. The vectors are supplied in three reading frames to facilitate in frame cloning, with a C-terminal peptide containing a polyhistidine metal-binding tag and the myc (c-myc) epitope. The human cytomegalovirus immediate-early (CMV) promoter provides highlevel expression in a wide range of mammalian cells. In addition, the vector will replicate episomally in cell lines that are latently infected with SV40 or that express the SV40 large T antigen (e.g. COS7). High-level stable and nonreplicative transient expression can be carried out in most mammalian cells. The control plasmid, pcDNA3.1/myc-His(-)/lacZ, is the pcDNA3.1/myc-His(-) A vector with a 3.2 kb fragment containing the β-galactosidase gene cloned in frame with the C-terminal peptide. It is included for use as a positive control for transfection, expression, and detection in the cell line of choice.

pCDNA3.1(-)/Myc-His B Sequence

LOCUS       Exported                5520 bp ds-DNA    circular SYN 29-10-2015
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5520)
  AUTHORS   admin
  TITLE     Direct Submission
  JOURNAL   Exported 2015-10-29  

FEATURES             Location/Qualifiers
     source          1..5520
                     /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"
     CDS             1005..1034
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /note="Myc"
                     /translation="EQKLISEEDL"
     CDS             1050..1067
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     polyA_signal    1117..1341
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1387..1815
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1829..2158
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      2009..2144
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             2225..3019
                     /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    3193..3314
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3363..3379)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    3387..3403
                     /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(3411..3441)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3456..3477
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3765..4353)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4524..5384)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
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