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 |
买家导航
pcDNA3.4载体 pcDNA3.4-TOPO质粒基本信息
pcDNA3.4质粒图谱 pcDNA3.4-TOPO载体图谱和pcDNA3.4载体序列 pcDNA3.4-TOPO质粒序列多克隆位点信息
pcDNA3.4质粒 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质粒序列 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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pcDNA3.4质粒 pcDNA3.4-TOPO载体应用举例
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