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pECMV-3×FLAG-N

pECMV-3×FLAG-N
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  • 产品名称：pECMV-3×FLAG-N
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  • 产品展商：XYbscience
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简单介绍
pECMV-3×FLAG-N菌株发货前均经过严格的多重验证，如存在质量问题，请在收到产品的三个月内通知我司。pECMV-3×FLAG-N收到质粒后请短暂离心，加入20μl ddH2O溶解质粒，取2μl转化至对应感受态中，挑取单克隆重新提取质粒后使用。
产品描述
 

pECMV-3×FLAG-N

  • 产品信息
  • 基本信息
  • 质粒简介
  • 质粒图谱
  • 质粒序列

产品信息

产品货号 产品名称 产品规格 优惠价
XY0159 pECMV-3×FLAG-N

5μg

¥请询价

使用说明

信裕质粒平台的各批次质粒菌株发货前均经过严格的多重验证，如存在质量问题，请在收到产品的三个月内通知我司。收到质粒后请短暂离心，加入20μl ddH2O溶解质粒，取2μl转化至对应感受态中，挑取单克隆重新提取质粒后使用。如需获取其他详细信息请登录我司官网查询。

基本信息

启动子: CMV promoter，T7 promoter
复制子: f1 ori，SV40 ori
终止子: SV40 poly(A) signal
质粒分类: 哺乳系列质粒；哺乳表达质粒；pCMV系列质粒
质粒大小: 5508bp
质粒标签: N-3×FLAG
原核抗性: 氨苄青霉素Amp
筛选标记: 新霉素Neo
克隆菌株: DH5α等哺乳细胞
培养条件: 37℃，有氧，LB
表达宿主: 293T等哺乳细胞
培养条件: 37℃，5%CO2
诱导方式: 无需诱导，瞬时表达
5'测序引物: pCDNA3.1-F（CTAGAGAACCCACTGCTTAC）
3'测序引物: pCDNA3.1-R（TAGAAGGCACAGTCGAGG）

质粒简介

        pECMV-3×FLAG-N含有一个在哺乳动物细胞中高水平表达的强启动子CMV，以及易于用抗体检测的3×FLAG标签，可用G418来筛选稳定细胞系，多克隆位点MCS处有丰富的酶切位点，可以用来连接目的片段，进而和3×FLAG融合表达。

质粒图谱



质粒序列

LOCUS       Exported                5502 bp ds-DNA     circular SYN 30-JUN-2017
DEFINITION  synthetic circular DNA
KEYWORDS    pECMV-3×FLAG-N
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5502)
  TITLE     Direct Submission
  JOURNAL   Exported Friday, June 30, 2017 from MiaoLingPlasmid
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..5502
                     /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             922..987
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /note="3xFLAG"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    985..1076
                     /note="MCS"
     polyA_signal    1102..1326
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1372..1800
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1814..2143
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      1994..2129
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             2210..3004
                     /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    3178..3299
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3348..3364)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    3372..3388
                     /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(3396..3426)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3441..3462
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3750..4335)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4506..5366)
                     /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(5367..5471)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt ggtaccgagc tcggatccac tagtccagtg
     1021 tggtggaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccgttt
     1081 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     1141 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     1201 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     1261 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     1321 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     1381 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     1441 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     1501 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     1561 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     1621 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     1681 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     1741 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     1801 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     1861 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     1921 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     1981 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2041 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     2101 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     2161 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     2221 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     2281 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     2341 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     2401 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     2461 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     2521 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     2581 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     2641 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     2701 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     2761 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     2821 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     2881 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     2941 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3001 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     3061 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     3121 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     3181 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3241 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3301 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     3361 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3421 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3481 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3541 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3601 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3661 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3721 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3781 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3841 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3901 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3961 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4021 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4081 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4141 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4201 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4261 ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga
     4321 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4381 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4441 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4501 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4561 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4621 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4681 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4741 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4801 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4861 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4921 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4981 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5041 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5101 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5161 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5221 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5281 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5341 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5401 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5461 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
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