pEGFP-Notch1-NICD-r大鼠基因质粒

保存条件 ：负20度

保质期 ：2年

供应商 ：钦诚生物

规格 ：5ug

基本信息

启动子:	CMV 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
培养条件:	5%CO2，37℃
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）

质粒简介

        pEGFP-Notch1-NICD-r质粒在CMV强启动子的作用下表达EGFP-Notch1-NICD融合蛋白，其中的跨膜大分子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质粒转染进COS-7L细胞，可以观察到绿色荧光蛋白为核定位，提示克隆的NICD基因符合进一步研究NICD在分裂后神经元的功能。
       

质粒图谱

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 
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