- 商品货号:FH1306
- 规格:10μl质粒
- 启动子:ECMV promoter
- 复制子:pUC ori,f1 ori,SV40 ori
- 终止子:bGH poly(A) signal
- 质粒分类:基因文库质粒;小鼠基因质粒;小鼠真核质粒
- 质粒大小:6100bp
- 质粒标签:N-3×FLAG
- 原核抗性:Amp(100μg/ml)
- 筛选标记:Neo
- 克隆菌株:DH5α
- 表达宿主:293T等哺乳细胞
- 诱导方式:无需诱导
- 5测序引物:CMV-F(CGCAAATGGGCGGTAGGCGTG)
- 3测序引物:BGH-R(TAGAAGGCACAGTCGAGG)
- 备注:本质粒由武汉淼灵生物科技有限公司分享至淼灵质粒平台。
- 质粒简介: pECMV-3×FLAG-HMGB1-m质粒是在pECMV-3×FLAG载体的基础上改造而成,CMV启动小鼠源HMGB1基因的过表达,N端的3×FLAG融合标签能很好的被FLAG抗体识别。该质粒可作为过表达质粒直接转染哺乳细胞。质粒本身带有Neo筛选标记,可用G418筛选转染后的细胞得到稳转细胞系。
基因NCBI信息:Mus musculus high mobility group box 1 (Hmgb1), transcript variant 1, mRNA;NCBI Reference Sequence: NM_001313894.1
质粒分享者信息:武汉淼灵生物科技有限公司成立于2014年6月,是专业从事生物技术服务及生化试剂研发、生产和销售的公司。我们致力于研发出质优价适的精品试剂,打破国内企业在低端恶性竞争、国外公司漫天要价的痛心局面,不断开拓国内外市场,成为中国生物公司里的海尔和华为,国外生物公司中的ATCC和addgene。
- 质粒图谱1: png/1-1FG01R045X9.png
质粒序列: 点击查看序列LOCUS Exported 6100 bp ds-DNA circular SYN 10-七月-2017
KEYWORDS pECMV-3×FLAG-HMGB1-m
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6100)
AUTHORS liutao
TITLE Direct Submission
JOURNAL Exported 2017年7月10日 from MiaoLingPlasmid
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6100
/organism="synthetic DNA construct"
/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"
CDS 922..987
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/note="3xFLAG"
/translation="DYKDHDGDYKDHDIDYKDDDDK"
misc_feature 1009..1656
/note="HMGB1-m"
polyA_signal 1700..1924
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 1970..2398
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2412..2741
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2592..2727
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 2808..3602
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="Neo"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3776..3897
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(4348..4933)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5104..5964)
/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(5965..6069)
/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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
961 atcgactaca aggatgacga tgacaagctt ggtaccgagc tcggatccat gggcaaagga
1021 gatcctaaaa agccgagagg caaaatgtcc tcatatgcat tctttgtgca aacttgccgg
1081 gaggagcaca agaagaagca cccggatgct tctgtcaact tctcagagtt ctccaagaag
1141 tgctcagaga ggtggaagac catgtctgct aaagaaaagg ggaaatttga agatatggca
1201 aaggctgaca aggctcgtta tgaaagagaa atgaaaacct acatcccccc caaaggggag
1261 accaaaaaga agttcaagga ccccaatgca cccaagaggc ctccttcggc cttcttcttg
1321 ttctgttctg agtaccgccc caaaatcaaa ggcgagcatc ctggcttatc cattggtgat
1381 gttgcaaaga aactaggaga gatgtggaac aacactgcag cagatgacaa gcagccctat
1441 gagaagaaag ctgccaagct gaaggagaag tatgagaagg atattgctgc ctacagagct
1501 aaaggaaaac ctgatgcagc gaaaaagggg gtggtcaagg ctgaaaagag caagaaaaag
1561 aaggaagagg aagatgatga ggaggatgaa gaggatgagg aagaggagga agaagaggaa
1621 gacgaagatg aagaagaaga tgatgatgat gaataactcg agtctagagg gcccgtttaa
1681 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
1741 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
1801 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
1861 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
1921 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
1981 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
2041 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
2101 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
2161 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
2221 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
2281 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
2341 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
2401 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
2461 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
2521 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
2581 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
2641 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
2701 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
2761 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
2821 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
2881 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
2941 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
3001 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
3061 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
3121 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
3181 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
3241 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
3301 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
3361 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
3421 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
3481 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
3541 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
3601 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
3661 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
3721 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
3781 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
3841 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
3901 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
3961 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
4021 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
4081 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
4141 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
4201 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
4261 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
4321 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
4381 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
4441 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
4501 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
4561 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
4621 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
4681 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
4741 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
4801 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
4861 caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa
4921 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
4981 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
5041 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
5101 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
5161 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
5221 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
5281 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
5341 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
5401 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
5461 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
5521 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
5581 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
5641 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
5701 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
5761 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
5821 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
5881 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
5941 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
6001 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
6061 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
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