p3xFLAG-CMV-14 载体 (V011361#)

载体基本信息
载体名称 p3xFLAG-CMV-14 载体抗性 Ampicillin
载体长度 6310 bp 载体类型 Mammalian Expression Vectors
载体来源 Sigma-Aldrich 筛选标记 Neomycin/G418(Geneticin)
拷贝数 High copy number 启动子 CMV
克隆方法 Enzyme digestion and ligation 5'引物 M13 fwd
3'引物 M13 rev

p3xFLAG-CMV-14 载体载体图谱

p3xFLAG-CMV-146310 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300M13 fwdCMV enhancerCMV promoterMCS3xFLAGhGH poly(A) signalSV40 promoterNeoR/KanRSV40 poly(A) signalM13 revoriAmpRAmpR promoterf1 ori

p3xFLAG-CMV-14 载体载体序列

LOCUS       Exported                6310 bp ds-DNA    circular SYN 28-11-2013
DEFINITION  Vector with a neomycin resistance marker for expression of 
            C-terminally 3xFLAG-tagged proteins in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    p3xFLAG-CMV-14
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6310)
  AUTHORS   Sigma-Aldrich
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6310
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     primer_bind     141..157
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    924..990
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             993..1058
                     /codon_start=1
                     /product="three tandem FLAG epitope tags, followed by an 
                     enterokinase cleavage site"
                     /note="3xFLAG"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1069..1691
                     /note="hGH poly(A) signal"
                     /note="human growth hormone polyadenylation signal"
     promoter        1733..2030
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             2084..2878
                     /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)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3530..3664
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3702..3718)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4104..4692)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4863..5723)
                     /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(5724..5828)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      5855..6310
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"

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