| 出品公司: | Invitrogen |
|---|---|
| 载体名称: | pcDNA4/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: | 限制性酶 |
| 启动子: | CMV |
| 载体大小: | 5096 dp |
| 5' 测序引物及序列: | 5'd[TAATACGACTCACTATAGGG]3' |
| 3' 测序引物及序列: | -- |
| 载体标签: | 6X His, V5 |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | 博来霉素 |
| 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: | V86120 |
| 稳定性: | 瞬时表达 |
| 组成型/诱导型: | -- |
| 病毒/非病毒: | 非病毒 |
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载体基本信息
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载体简介
载体序列
LOCUS pcDNA4 V5-His B 5096 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)4/V5-His A or
pcDNA(TM)4/V5-His C.
ACCESSION .
VERSION .
KEYWORDS pcDNA4 V5-His B
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5096)
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..5096
/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 895..1012
/note="MCS"
/note="multiple cloning site"
CDS 1013..1054
/codon_start=1
/product="epitope tag from simian virus 5"
/note="V5 tag"
/translation="GKPIPNPLLGLDST"
CDS 1064..1081
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
polyA_signal 1110..1334
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 1380..1808
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1822..2151
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2002..2137
/note="SV40 ori"
/note="SV40 origin of replication"
promoter 2199..2246
/note="EM7 promoter"
/note="synthetic bacterial promoter "
CDS 2265..2639
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/note="BleoR"
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 2769..2890
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(3341..3929)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4100..4960)
/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(4961..5065)
/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 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc
961 agatatccag cacagtggcg gccgctcgag tctagagggc ccgcggttcg aaggtaagcc
1021 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca
1081 ttgagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
1141 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
1201 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
1261 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
1321 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
1381 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
1441 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
1501 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
1561 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
1621 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
1681 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
1741 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
1801 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
1861 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
1921 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
1981 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
2041 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
2101 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
2161 ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat
2221 agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca
2281 gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc
2341 ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg
2401 tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg
2461 tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact
2521 tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt
2581 tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac
2641 acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
2701 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
2761 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
2821 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
2881 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
2941 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
3001 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
3061 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
3121 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
3181 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
3241 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
3301 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
3361 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
3421 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
3481 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
3541 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
3601 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
3661 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
3721 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
3781 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
3841 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
3901 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
3961 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
4021 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
4081 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
4141 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
4201 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
4261 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
4321 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
4381 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
4441 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
4501 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
4561 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
4621 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
4681 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
4741 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
4801 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
4861 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
4921 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
4981 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
5041 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
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