pCMV- Tag 4B/ pCMV- Tag 4B质粒/ pCMV- Tag 4B Plasmid

PVT1029 2ug

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pCMV-Tag 4B Information

Promoter: CMV

Replicon: pUC ori, F1 ori

Terminator: SV40 poly (A) signal

Plasmid Classification: Mammalian Cell Vector; Protein Overexpression Plasmid

Plasmid size: 4320 BP

Plasmid tag: C-FLAG tag

Prokaryotic resistance: kanamycin Kan

Screening marker: neomycin Neo

Cloning strain: XL1-Blue

Culture conditions: 37 C, aerobic, LB

Expression host: mammalian cells

Culture conditions: 37 C, aerobic, LB

Induction mode: instantaneous expression without induction

5'Sequencing Primers: CMV-F: CGCAAATGGGGGGGTAGGGGGGGTG

3'Sequencing Primers: Sequence-based Design

pCMV-Tag 4B Sites

pCMV-Tag 4B Sequence

LOCUS       Exported                4320 bp ds-DNA     circular SYN 28-Nov-2013
DEFINITION  Mammalian expression vector for tagging proteins with a C-terminal
            FLAG epitope. For other reading frames, use pCMV-Tag 4A or pCMV-Tag
            4C.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-Tag 4B
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4320)
  AUTHORS   Agilent Technologies
  TITLE     Direct Submission
  JOURNAL   Exported 2016-09-18 from SnapGene Viewer 2.8.1
             
COMMENT     The cloned gene must include a Kozak sequence and start codon.
FEATURES             Location/Qualifiers
     source          1..4320
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     enhancer        66..370
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        620..638
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    651..744
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             745..768
                     /codon_start=1
                     /product="FLAG epitope tag, followed by an enterokinase
                     cleavage site"
                     /note="FLAG"
                     /translation="DYKDDDDK"
     misc_feature    784..794
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     promoter        complement(822..840)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1114..1235
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1242..1697)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1724..1826
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        1830..2187
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             2222..3016
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    3248..3295
                     /note="HSV TK poly(A) signal"
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     rep_origin      3624..4212
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
      661 gcggtggcgg ccgctctagc ccgggcggat cccccgggct gcaggaattc gatatcaagc
      721 ttatcgatac cgtcgacact cgaggattac aaggatgacg acgataagta gggcccggta
      781 ccttaattaa ttaaggtacc aggtaagtgt acccaattcg ccctatagtg agtcgtatta
      841 caattcactc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggagatccaa
      901 tttttaagtg tataatgtgt taaactactg attctaattg tttgtgtatt ttagattcac
      961 agtcccaagg ctcatttcag gcccctcagt cctcacagtc tgttcatgat cataatcagc
     1021 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     1081 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     1141 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1201 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     1261 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     1321 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     1381 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     1441 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     1501 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     1561 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     1621 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     1681 gccgct <