pEGFP-Notch1-NICD-r BioVector NTCC质粒载体菌种细胞基因保藏中心

pEGFP-Notch1-NICD-r

BioVector NTCC质粒载体菌种细胞基因保藏中心                      

启动子: ECMV promoter
复制子: pUC ori,F1 ori,SV40 ori
终止子: SV40 poly(A) signal
质粒分类: 基因文库质粒；大鼠基因质粒；大鼠真核质粒
质粒大小: 7079bp
质粒标签: C-EGFP
原核抗性: Kan（50μg/ml）
筛选标记: Neo/G418
克隆菌株: DH5α等大肠杆菌
培养条件: 37℃，LB，有氧
表达宿主: 293T等哺乳细胞
诱导方式: 无需诱导
5'测序引物: pEGFP-N-5（TGGGAGGTCTATATAAGCAGAG）
3'测序引物: pEGFP-N-3（CGTCGCCGTCCAGCTCGACCAG）

Map图谱

质粒简介
       

跨膜大分子Notch1受体主要参与发育过程中的细胞命运的选择。其G1743和V1744部位发生水解释放产生的活性片段－－胞质段（Notch1 intracellular cytoplasmic domain，NICD）能被转运进核，激活下激靶基因的表达，Notch1参与细胞的增殖、分化、程序性死亡、发育过程中的形态发生和器官形成等许多重要过程。我们构建的pEGFP-Notch1-NICD-r质粒转染COS-7L，观察到绿色荧光蛋白为核定位，提示克隆的NICD符合进一步研究NICD在分裂后神经元的功能。

质粒序列

LOCUS       Exported                7097 bp ds-DNA     circular SYN 04-JUL-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pEGFP-Notch1-NICD-r
SOURCE      synthetic DNA construct
 ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7097)
 AUTHORS   yinchu
 TITLE     Direct Submission
 JOURNAL   Exported Tuesday, July 4, 2017 from SnapGene Viewer 3.3.4
           http://www.snapgene.com
FEATURES             Location/Qualifiers
    source          1..7097
                    /organism="synthetic DNA construct"
                    /mol_type="other DNA"
    enhancer        61..364
                    /label=CMV enhancer
                    /note="human cytomegalovirus immediate early enhancer"
    promoter        365..568
                    /label=CMV promoter
                    /note="human cytomegalovirus (CMV) immediate early
                    promoter"
    misc_feature    637..3002
                    /label=Notch1-NICD-r
                    /note="Rattus norvegicus notch 1 (Notch1), mRNA

                    NCBI Reference Sequence: NM_001105721.1

                    "
    CDS             3043..3762
                    /codon_start=1
                    /product="enhanced GFP"
                    /label=EGFP
                    /note="mammalian codon-optimized"
                    /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                    KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                    GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                    VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                    EFVTAAGITLGMDELYK"
    polyA_signal    3885..4006
                    /label=SV40 poly(A) signal
                    /note="SV40 polyadenylation signal"
    rep_origin      complement(4013..4468)
                    /direction=LEFT
                    /label=f1 ori
                    /note="f1 bacteriophage origin of replication; arrow
                    indicates direction of (+) strand synthesis"
    promoter        4495..4599
                    /gene="bla"
                    /label=AmpR promoter
    promoter        4601..4958
                    /label=SV40 promoter
                    /note="SV40 enhancer and early promoter"
    rep_origin      4809..4944
                    /label=SV40 ori
                    /note="SV40 origin of replication"
    CDS             4993..5787
                    /codon_start=1
                    /gene="aph(3')-II (or nptII)"
                    /product="aminoglycoside phosphotransferase from Tn5"
                    /label=NeoR/KanR
                    /note="confers resistance to neomycin, kanamycin, and G418
                    (Geneticin(R))"
                    /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                    VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                    SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                    GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                    LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
    polyA_signal    6019..6066
                    /label=HSV TK poly(A) signal
                    /note="herpes simplex virus thymidine kinase
                    polyadenylation signal (Cole and Stacy, 1985)"
    rep_origin      6395..6983
                    /direction=RIGHT
                    /label=ori
                    /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                    replication"
ORIGIN
       1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
     601 ccggactcag atctcgagct caagcttcga attctgatgc tgctgtcccg caagcgcagg
     661 cggcagcatg gccagctctg gttccctgag ggtttcaaag tgtcagaggc cagcaagaag
     721 aagcggagag aacccctcgg cgaggactca gtcggcctca agcccctgaa gaacgcctca
     781 gatggtgccc tgatggacga caatcagaac gagtgggggg acgaagacct ggagaccaag
     841 aagttccggt ttgaggaacc agtggttctc cctgacctgg atgatcagac tgaccaccgg
     901 cagtggaccc agcagcacct ggatgccgct gacctacgtg tgtctgccat ggccccaacg
     961 ccgcctcagg gggaggtaga tgctgactgc atggacgtca atgttcgagg accagatggc
    1021 ttcactcccc taatgattgc ctcctgcagc ggagggggcc tggagacagg caacagtgag
    1081 gaagaagaag atgcacctgc tgtcatctcc gacttcatct atcagggtgc cagcttgcac
    1141 aaccagacgg accgcacagg ggagactgcc ctgcacctgg ctgcccgata ctctcgttca
    1201 gatgctgcca agcgcttgct ggaggccagc gcagatgcca acatccaaga caacatgggg
    1261 cgtaccccat tacatgccgc tgtttctgca gacgctcagg gtgtcttcca gatcctgctc
    1321 cggaacagag ccacagatct ggatgcccga atgcatgatg gcacaacccc tctgatcctg
    1381 gcggcacgcc tggccgtgga aggcatgcta gaggacctca tcaactctca cgctgatgtc
    1441 aatgctgtgg atgacctagg caagtcagct ctgcactggg cagccgctgt gaacaatgtg
    1501 gacgctgctg ttgtgctcct gaagaacgga gccaacaaag acatgcagaa caacaaggag
    1561 gagactcccc tgttcctggc cgcccgtgag ggcagctatg agactgccaa agtgttgctg
    1621 gaccactttg ccaaccggga catcacggat cacatggacc gattgccacg ggacattgca
    1681 caggagcgca tgcaccacga tatcgtgcgg cttttggatg aatacaacct ggtgcgcagc
    1741 ccacagctgc atggcactgc cttgggtggc acacccactc tgtctcccac actctgctcg
    1801 cccaacggct acctgggcaa cctcaagtct gccacacagg gcaagaaggc ccgaaagccc
    1861 agcaccaaag ggctggcttg cagtagcaag gaagctaagg acctcaaggc ccggaggaag
    1921 aagtcccagg atggcaaggg ctgcctgttg gacagctcaa gcatgctgtc acccgtggac
    1981 tccctcgagt caccccatgg ctacttgtca gatgtggcct caccacccct ccttccctcc
    2041 ccgttccagc agtctccatc catgcctctc agccacctgc caggtatgcc tgacacccac
    2101 ctgggcatca gccacttgaa tgtggcagcc aagcccgaga tggcagctct ggccggaggc
    2161 agccggttgg cctttgagcc acccccacca cgcctctccc acctgcctgt agcctccagt
    2221 gccagcacag tgctgagtac caatggcaca ggggctatga atttcaccgt gggtgcaccg
    2281 gcaagcttga atggccagtg tgagtggctt ccccggctcc agaatggcat ggtgcccagc
    2341 cagtacaacc cgctaaggcc aggtgtgact ccgggcacac taagcacaca ggcagctggc
    2401 ctccagcatg gcatgatggg cccgatacac agcagcctct ccaccaatac cttgtccccg
    2461 attatctacc agggcctgcc caacacaagg ctggccacac agccccacct ggtgcagacc
    2521 cagcaggtgc agccacagaa cttacaaatc cagcctcaga acctgcagcc accatcgcag
    2581 ccacacctca gtgtgagctc agcagccaat gggcacctgg gtcggagctt cctgagcggg
    2641 gagcccagcc aggcagacgt acagccgctg ggccccagca gtctgcctgt gcacaccatt
    2701 ctgccccagg aaagccaggc tctgccgaca tcactgccat cctccatggt cccacccatg
    2761 accactaccc agttcctgac ccctccttct cagcacagct actcatcctc acctgtggac
    2821 aacaccccca gccaccagct gcaggtgcca gagcacccct tcctcacccc atcccctgag
    2881 tcccctgacc agtggtccag ctcctccccg cattccaaca tctctgattg gtccgagggc
    2941 atctctagcc cgcccacgag catgccgtcc cagatcaccc acattccaga ggcatttaag
    3001 tcgtcgacgg taccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag
    3061 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
    3121 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
    3181 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
    3241 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
    3301 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
    3361 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
    3421 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
    3481 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
    3541 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
    3601 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
    3661 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
    3721 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga
    3781 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
    3841 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
    3901 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
    3961 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
    4021 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
    4081 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
    4141 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
    4201 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
    4261 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
    4321 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
    4381 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
    4441 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
    4501 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
    4561 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
    4621 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
    4681 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
    4741 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
    4801 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
    4861 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
    4921 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
    4981 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
    5041 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
    5101 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
    5161 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
    5221 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
    5281 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
    5341 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
    5401 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
    5461 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
    5521 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
    5581 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
    5641 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
    5701 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
    5761 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
    5821 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
    5881 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
    5941 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
    6001 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
    6061 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
    6121 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
    6181 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
    6241 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
    6301 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
    6361 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
    6421 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
    6481 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
    6541 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
    6601 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
    6661 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
    6721 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
    6781 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
    6841 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
    6901 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
    6961 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
    7021 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
    7081 gtattaccgc catgcat
//      [Supplier来源]  http://www.biovector.net