质粒名称: | pEGFP-rNFM |
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出品公司: | Addgene |
目录编号: | 32907 |
存储实验室: | Anthony Brown |
载体骨架基本信息 |
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载体名称: | pEGFP-C1 |
载体出品公司: | Clontech |
载体抗性: | 卡那霉素 |
空载体大小: | 4134 bp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳细胞表达 |
筛选标记: | 新霉素(G418) |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | EGFP-rat NFM fusion,NFM,neurofilament protein M |
物种来源: | R. norvegicus (rat) |
插入大小: | 3330 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | CMV |
标签/融合蛋白: | EGFP(插入端N端) |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | ApaI(未破坏) |
3' 克隆位点: | ApaI(克隆过程中销毁) |
5' 测序引物 | -- |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pEGFP-rNFM was a gift from Anthony Brown (Addgene plasmid # 32907 ; http://n2t.net/addgene:32907 ; RRID:Addgene_32907) |
参考文献部分: | Rapid movement of axonal neurofilaments interrupted by prolonged pauses. Wang L, Ho CL, Sun D, Liem RK, Brown A. Nat Cell Biol. 2000 Mar;2(3):137-41. 10.1038/35004008 PubMed 10707083 |
买家导航
pEGFPrNFM载体基本信息
pEGFPrNFM载体图谱质粒图谱和多克隆位点信息
pEGFPrNFM载体简介
pEGFPrNFM载体序列
LOCUS pEGFP-rNFM 7464 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Mammalian expression construct driving the expression of a fluorescent fusion protein consisting of EGFP linked to the N-terminus of rat neurofilament protein M. ACCESSION . VERSION . KEYWORDS pEGFP-rNFM SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7464) AUTHORS Wang L, Ho CL, Sun D, Liem RK, Brown A TITLE Rapid movement of axonal neurofilaments interrupted by prolonged pauses. JOURNAL Nat Cell Biol. 2000 Mar;2(3):137-41. PUBMED 10707083 REFERENCE 2 (bases 1 to 7464) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7464 /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" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1329 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(658..679) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(919..938) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1266..1287 /label=EGFP-C /note="EGFP, forward primer" RBS 2854..2865 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" polyA_signal 4252..4373 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4289..4308) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4343..4362 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4380..4835) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4517..4538) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4729..4748 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4862..4966 /gene="bla" /label=AmpR promoter promoter 4968..5325 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4991..5011) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5176..5311 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5238..5257 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5360..6154 /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" primer_bind complement(5414..5433) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6024..6043 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(6342..6361) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6386..6433 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6762..7350 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7251..7270 /label=pBR322ori-F /note="pBR322 origin, forward primer" 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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc 841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac 1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc 1381 gcgggcccaa ttccggcctc caagatgagc tacacgctgg actcgctggg caacccgtcc 1441 gcctaccggc gggtcaccga gacccgctcc agcttcagtc gtgtgagcgg ttccccgtcc 1501 agcggcttcc gctcgcagtc ctggtcccgc ggctcgccca gcaccgtgtc ctcctcctac 1561 aagcgcagcg cgctcgcccc gcgcctcgcc tacagctcgg ctatgctcag ctcggccgag 1621 agcagcctcg acttcagcca gtcctcttcg ctgcttaacg gcggctccgg cggcgactac 1681 aagctgtccc gctccaacga gaaagagcag ctgcaggggc tgaacgaccg cttcgccggc 1741 tacatcgaga aagtgcacta cttggaacaa cagaacaagg agatcgaggc agagatccac 1801 gcgctgcggc agaagcaggc ctcgcacgcc cagctgggtg acgcttacga ccaggagatc 1861 cgagagctgc gcgccaccct ggagatggtg aatcacgaga aggctcaagt gcagctggac 1921 tctgatcact tggaggaaga catccaccgg ctcaaggagc gcttcgagga ggaggcgcgg 1981 ctgcgggacg acaccgaggc tgccatccgg gcgctgcgca aagacataga ggagtcgtcg 2041 atggttaagg tggagctgga caagaaggtg cagtcgctgc aggatgaggt ggccttcctg 2101 cggagcaatc acgaagagga ggtggccgac ctgctggccc agatccaggc gtcgcacatc 2161 accgtagagc gcaaagacta cctgaagaca gacatctcca cggcgctgaa agagatccgc 2221 tcccagctcg agtgtcactc cgaccagaac atgcaccagg ccgaagagtg gttcaaatgc 2281 cgctacgcca agctcaccga ggcggccgag cagaacaagg aggccatccg ctccgctaaa 2341 gaagagatcg ccgagtaccg gcgccagctg cagtccaaga gcattgagct cgagtcggtg 2401 cgaggcacta aggagtccct ggaacggcag ctcagcgaca tcgaggagcg ccacaaccac 2461 gacctcagca gctaccagga caccatccag cagctggaaa atgagcttcg gggaacaaag 2521 tgggaaatgg ctcgtcattt gcgagaatac caggatctcc ttaacgtcaa gatggctctg 2581 gacatcgaga tcgccgcata taggaaacta ctggagggtg aagagaccag atttagcaca 2641 ttttcaggaa gcatcactgg gcctctgtac acacaccgac agccctcagt cacaatatcc 2701 agtaagattc agaagaccaa agtcgaggcc cccaagctca aggtccaaca caaatttgtg 2761 gaggagatca ttgaggagac taaagtggaa gatgagaagt cagaaatgga agacgccctc 2821 acagtcattg cagaggaatt ggcagcctct gccaaagagg agaaagaaga ggcagaagaa 2881 aaggaagagg aaccggaagt tgaaaagtct cccgtgaagt ctcctgaggc taaggaagag 2941 gaggaagggg aaaaggagga agaagaggaa ggccaagagg aagaagagga ggaagatgaa 3001 ggtgtcaagt cagaccaggc agaagaggga ggatctgaga aggaaggctc gagtgaaaag 3061 gatgaaggtg agcaagaaga agaaggggaa actgaggcag aaggtgaagg agaggaagca 3121 gaagctaagg aggaaaagaa aacagaggga aaggtcgagg aaatggctat caaggaggaa 3181 atcaaggtcg agaagcccga gaaagccaag tcccctgtgc caaaatcacc cgtggaagaa 3241 gtaaagccaa aaccagaagc caaagccgga aaggatgagc agaaggagga agagaaagtt 3301 gaggagaaga aggaggtagc caaggaatca cccaaggaag agaaggtgga gaaaaaggag 3361 gagaagccaa aagatgtccc agataaaaag aaggctgagt ccccagtgaa agaaaaggcc 3421 gtagaggaaa tgatcaccat tactaagtcg gtaaaggtga gcctggagaa agacaccaaa 3481 gaggagaagc ctcagcagca ggagaaggtg aaggagaagg cagaggagga ggggggtagt 3541 gaggaggaag tgggtgacaa aagcccgcaa gaatccaaga aggaagacat agctatcaat 3601 ggggaggtgg aaggaaaaga ggaggaggag caggaaactc aggagaaggg cagtgggcaa 3661 gaggaggaga aaggggtggt cactaatggc ttagatgtga gccctgcgga ggaaaagaaa 3721 ggggaggata gaagtgatga caaagtggtg gtgaccaaga aggtagaaaa aatcaccagc 3781 gagggaggcg atggtgctac caaatacatc accaaatctg ttactgtcac tcaaaaggtt 3841 gaagagcatg aggagacctt tgaggagaag ctggtgtcaa ctaaaaaggt agaaaaggtc 3901 acttcacatg ccatagtcaa ggaagtcacc cagggtgact aagatcggag tccattgcaa 3961 aaggttaagc catacaacaa tttcaaaatg catgtgattg acagcttcaa aacagaatgg 4021 gttctcccat gagggctcca gacattgtat tttcctttgt gcaatatgag ggaactgcat 4081 gcaagctcag ggtgcccccc tcctcagtcc ttgggggaat tgggatccac cggatctaga 4141 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc 4201 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt 4261 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 4321 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta 4381 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt 4441 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat 4501 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa 4561 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta 4621 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc 4681 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc 4741 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac 4801 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt 4861 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag 4921 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag 4981 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 5041 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 5101 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 5161 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 5221 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc 5281 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac 5341 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc 5401 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc 5461 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc 5521 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg 5581 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt 5641 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc 5701 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga 5761 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga 5821 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct 5881 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc 5941 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt 6001 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg 6061 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat 6121 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc 6181 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa 6241 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 6301 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca 6361 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt 6421 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca 6481 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca 6541 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc 6601 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 6661 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 6721 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 6781 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 6841 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 6901 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 6961 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 7021 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 7081 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 7141 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 7201 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 7261 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 7321 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 7381 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg 7441 gataaccgta ttaccgccat gcat //
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