• 规格：20μl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
//