pCMV-Tag 4A哺乳表达质粒

基本信息

原核抗性:	Kan
筛选标记:	Neo/G418
克隆菌株:	DH5α
培养条件:	LB/37℃

质粒简介

pCMV-Tag 4A哺乳表达质粒

质粒图谱

质粒序列

LOCUS       Exported                4319 bp ds-DNA     circular SYN 10-JUL-2018
DEFINITION  Mammalian expression vector for tagging proteins with a C-terminal 
            FLAG epitope. For other reading frames, use pCMV-Tag 4B or pCMV-Tag 
            4C.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-Tag 4A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4319)
  AUTHORS   Agilent Technologies
  TITLE     Direct Submission
COMMENT     The cloned gene must include a Kozak sequence and start codon.
FEATURES             Location/Qualifiers
     source          1..4319
                     /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..743
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             744..767
                     /codon_start=1
                     /product="FLAG epitope tag, followed by an enterokinase 
                     cleavage site"
                     /note="FLAG"
                     /translation="DYKDDDDK"
     misc_feature    783..793
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     promoter        complement(821..839)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1113..1234
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1241..1696)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1723..1825
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        1829..2186
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             2221..3015
                     /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    3247..3294
                     /note="HSV TK poly(A) signal"
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     rep_origin      3623..4211
                     /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 cgtcgacctc gaggattaca aggatgacga cgataagtag ggcccggtac
      781 cttaattaat taaggtacca ggtaagtgta cccaattcgc cctatagtga gtcgtattac
      841 aattcactcg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gagatccaat
      901 ttttaagtgt ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattcaca
      961 gtcccaaggc tcatttcagg cccctcagtc ctcacagtct gttcatgatc ataatcagcc
     1021 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     1081 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     1141 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     1201 gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt
     1261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     1321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     1381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1561 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     1621 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     1681 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1741 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1801 cttcaataat attgaaaaag gaagaatcct gaggcggaaa gaaccagctg tggaatgtgt
     1861 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     1921 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     1981 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2041 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     2101 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     2161 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     2221 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     2281 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     2341 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     2401 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     2461 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     2521 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     2581 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     2641 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     2701 gaacatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     2761 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     2821 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     2881 atagcgttgg ctacccgtga tattgctgaa gaacttggcg gcgaatgggc tgaccgcttc
     2941 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     3001 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     3061 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     3121 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     3181 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     3241 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     3301 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     3361 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     3421 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     3481 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3541 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3601 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3661 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3721 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3781 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3841 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3901 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3961 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4021 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4081 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4141 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4201 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4261 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcc