pcDNA3.4

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联系方式:I47-825O-882O

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pcDNA3.4-TOPO载体基本信息

出品公司: Thermo fisher
载体名称: pcDNA3.4 , pcDNA3.4-TOPO ,pcDNA3.4载体 , pcDNA3.4-TOPO载体 
质粒类型: 哺乳动物细胞载体, 瞬时表达载体
高拷贝/低拷贝: 高拷贝
克隆方法: 限制性内切酶,多克隆位点
启动子: CMV
载体大小: 6.0 kb
5' 测序引物及序列: CMV-F: 5’–CGC AAA TGG GCG GTA GGC GTG–3’
3' 测序引物及序列: pcDNA3.4-R:5’–CAA CAT AGT TAA GAA TAC CAG TC–3’
载体标签: --
载体抗性: 氨苄青霉素
筛选标记: G418
克隆菌株: DH5α 等
宿主细胞(系): 常规细胞系(293、CV-1、CHO等)
备注: 用于蛋白的瞬转表达,表达量较高!
产品目录号: A14697
稳定性: 瞬时表达
组成型/诱导型: 组成型
病毒/非病毒: 非病毒

pcDNA3.4-TOPO载体质粒图谱和多克隆位点信息

pcDNA3.4载体图谱


pcDNA3.4多克隆位点

pcDNA3.4-TOPO载体简介

pcDNA3.4载体含有CMV启动子,用于哺乳细胞的蛋白高表达,或者哺乳细胞分泌性蛋白表达,使用瞬时转染方法进行蛋白表达。

pcDNA3.4载体含有的关键元件如下:
1、WPRE(Woodchuck posttranscriptional regulatory element)元件,位于多克隆位点下游,可以有效提高基因的转录和表达

 
2、CMV启动子能够增强多种基因在哺乳细胞中的高水平表达。
 
3、Herpes Simplex Virus thymidine kinase polyadenylation signal 单纯疱疹病毒胸苷激酶聚腺苷酸信号可以控制基因转录的终止。
 
4、拥有新霉素抗性基因,可以利用Geneticin筛选稳定表达细胞系。
 
5、pUC起始复制子,保证了质粒在大肠杆菌中的高拷贝数
 
6、在大肠杆菌中,具有氨苄抗性基因。

pcDNA3.4-TOPO载体序列

LOCUS       pcDNA3.4-TOPO                6011 bp ds-DNA     linear   SYN 02-MAY-2019
DEFINITION  Linearized mammalian vector with 3'-T overhangs and bound 
            topoisomerase, for TOPO(R) TA cloning of PCR products and high level
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.4-TOPO (linearized)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6011)
  AUTHORS   Thermo Fisher (Invitrogen)
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, January 2, 2020 from SnapGene Viewer 3.2.1
            http://www.snapgene.com
COMMENT     Do not use phosphorylated primers for the PCR amplification because 
            5' phosphates will prevent ligation into this vector.
FEATURES             Location/Qualifiers
     source          1..6011
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     misc_feature    46..634
                     /note="WPRE"
                     /note="woodchuck posttranscriptional regulatory element"
     polyA_signal    700..748
                     /note="HSV TK poly(A) signal"
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     rep_origin      950..1378
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1392..1721
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      1572..1707
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             1788..2582
                     /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    2758..2879
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3330..3918)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4089..4949)
                     /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(4950..5054)
                     /gene="bla"
                     /note="AmpR promoter"
     enhancer        5320..5699
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5700..5903
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 aagggttcga tccctaccgg ttagtaatga gtttgatatc tcgacaatca acctctggat
       61 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
      121 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
      181 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
      241 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
      301 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
      361 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
      421 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
      481 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
      541 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
      601 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa
      661 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
      721 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
      781 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
      841 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
      901 ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt
      961 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     1021 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     1081 tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg
     1141 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     1201 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     1261 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     1321 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     1381 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     1441 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     1501 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     1561 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     1621 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     1681 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     1741 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     1801 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     1861 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     1921 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     1981 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     2041 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     2101 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     2161 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     2221 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     2281 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     2341 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     2401 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     2461 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     2521 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     2581 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     2641 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     2701 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     2761 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2821 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     2881 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     2941 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3001 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3061 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3121 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3181 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3241 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3301 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3361 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3421 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3481 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     3541 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3601 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3661 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3721 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     3781 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3841 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3901 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3961 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4021 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4081 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4141 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4201 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4261 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4321 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4381 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4441 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4501 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4561 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4621 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4681 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4741 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4801 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4861 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4921 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4981 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5041 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     5101 ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc
     5161 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     5221 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     5281 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     5341 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     5401 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     5461 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     5521 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     5581 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     5641 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     5701 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     5761 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     5821 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     5881 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
     5941 cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg actctagagg
     6001 atcgaaccct t
//

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