| 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-TOPO载体基本信息
pcDNA3.4-TOPO载体图谱质粒图谱和多克隆位点信息
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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