| 出品公司: | Invitrogen |
|---|---|
| 载体名称: | pcDNA3.1/V5-His B |
| 质粒类型: | 哺乳动物细胞载体 |
| 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: | 5506 dp |
| 5' 测序引物及序列: | CMV-F |
| 3' 测序引物及序列: | BGH-rev |
| 载体标签: | His Tag (6x), V5 Epitope Tag |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | 新霉素 |
| 克隆菌株: | -- |
| 宿主细胞（系）: | -- |
| 备注: | 哺乳动物细胞表达C末端V5和6xHis标记蛋白的载体。 |
| 产品目录号: | -- |
| 稳定性: | -- |
| 组成型/诱导型: | -- |
| 病毒/非病毒: | 非病毒 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pcDNA3.1/V5-His B 5506 bp ds-DNA circular SYN 24-OCT-2017
DEFINITION Mammalian vector for expressing C-terminally V5- and 6xHis-tagged
proteins. For other reading frames, use pcDNA(TM)3.1/V5-His A or
pcDNA(TM)3.1/V5-His C.
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1 V5-His B
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5506)
AUTHORS Thermo Fisher (Invitrogen)
TITLE Direct Submission
JOURNAL Exported Saturday, November 16, 2019 from SnapGene Viewer 3.1.4
http://www.snapgene.com
COMMENT The cloned gene must include a Kozak sequence and start codon.
FEATURES Location/Qualifiers
source 1..5506
/organism="synthetic DNA construct"
/lab_host="Mammalian Cells"
/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"
misc_feature 902..1003
/note="MCS"
/note="multiple cloning site"
CDS 1004..1045
/codon_start=1
/product="epitope tag from simian virus 5"
/note="V5 tag"
/translation="GKPIPNPLLGLDST"
CDS 1055..1072
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
polyA_signal 1101..1325
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 1371..1799
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1813..2142
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 1993..2128
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 2209..3003
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3179..3300
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(3751..4339)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4510..5370)
/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(5371..5475)
/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 gctggctagt
901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcca
961 gcacagtggc ggccgctcga gtctagaggg cccgcggttc gaaggtaagc ctatccctaa
1021 ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgagttta
1081 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
1141 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
1201 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
1261 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
1321 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
1381 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
1441 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
1501 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
1561 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
1621 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
1681 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
1741 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
1801 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
1861 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
1921 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
1981 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
2041 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
2101 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
2161 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
2221 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
2281 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
2341 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
2401 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
2461 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
2521 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
2581 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
2641 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
2701 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
2761 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
2821 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
2881 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
2941 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
3001 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
3061 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
3121 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
3181 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
3241 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
3301 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
3361 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
3421 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
3481 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
3541 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
3601 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
3661 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
3721 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
3781 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
3841 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
3901 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
3961 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
4021 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
4081 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
4141 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
4201 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
4261 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
4321 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
4381 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
4441 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
4501 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
4561 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
4621 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
4681 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
4741 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
4801 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
4861 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
4921 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
4981 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
5041 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
5101 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
5161 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
5221 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
5281 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
5341 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
5401 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
5461 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
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