| 质粒名称: | pEGFP-C1-Ataxin3Q84 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 22123 |
| 存储实验室: | Henry Paulson |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-C1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4731 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）,G418 |
| 载体抗性: | 卡那霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Ataxin 3 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1300 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | EGFP（主干上的N端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamHI（如果已销毁，则未知） |
| 3’克隆位点: | BamHI（如果已销毁，则未知） |
| 5’测序引物 | EGFP-C |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pEGFP-C1-Ataxin3Q84 was a gift from Henry Paulson (Addgene plasmid # 22123 ; http://n2t.net/addgene:22123 ; RRID:Addgene_22123) |
| 参考文献部分: | Live-cell imaging reveals divergent intracellular dynamics of polyglutamine disease proteins and supports a sequestration model of pathogenesis. Chai Y, Shao J, Miller VM, Williams A, Paulson HL. Proc Natl Acad Sci U S A. 2002 Jul 9. 99(14):9310-5. 10.1073/pnas.152101299 PubMed 12084819 |
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载体基本信息
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载体序列
LOCUS pEGFP-C1-Ataxin3Q84 6004 bp ds-DNA circular SYN 16-AUG-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pEGFP-C1-Ataxin3Q84 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6004) AUTHORS Chai Y, Shao J, Miller VM, Williams A, Paulson HL TITLE Live-cell imaging reveals divergent intracellular dynamics of polyglutamine disease proteins and supports a sequestration model of pathogenesis. JOURNAL Proc Natl Acad Sci U S A. 2002 Jul 9. 99(14):9310-5. PUBMED 12084819 REFERENCE 2 (bases 1 to 6004) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 16, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6004 /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 /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 613..1329 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /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" misc_feature 1330..1395 /label=MCS /note="multiple cloning site" polyA_signal 2792..2913 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2829..2848) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2883..2902 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2920..3375) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3057..3078) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3269..3288 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3402..3506 /gene="bla" /label=AmpR promoter promoter 3508..3865 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3531..3551) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3716..3851 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3778..3797 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3900..4694 /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(3954..3973) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4564..4583 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4882..4901) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4926..4973 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5302..5890 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5791..5810 /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 gcgggcccgg gatccataaa catggagtcc atcttccacg agaaacaaga aggctcactt 1441 tgtgctcaac attgcctgaa taacttattg caaggagaat attttagccc tgtggaatta 1501 tcctcaattg cacatcagct ggatgaggag gagaggatga gaatggcaga aggaggagtt 1561 actagtgaag attatcgcac gtttttacag cagccttctg gaaatatgga tgacagtggt 1621 tttttctcta ttcaggttat aagcaatgcc ttgaaagttt ggggtttaga actaatcctg 1681 ttcaacagtc cagagtatca gaggctcagg atcgatccta taaatgaaag atcatttata 1741 tgcaattata aggaacactg gtttacagtt agaaaattag gaaaacagtg gtttaacttg 1801 aattctctct tgacgggtcc agaattaata tcagatacat atcttgcact tttcttggct 1861 caattacaac aggaaggtta ttctatattt gttgttaagg gtgatctgcc agattgcgaa 1921 gctgaccaac tcctgcagat gattagggtc caacagatgc atcgaccaaa acttattgga 1981 gaagaattag cacaactaaa agagcaaaga gtccataaaa cagacctgga acgaatgtta 2041 gaagcaaatg atggctcagg aatgttagac gaagatgagg aggatttgca gagggctctg 2101 gcactaagtc gccaagaaat tgacatggaa gatgaggaag cagatctccg cagggctatt 2161 cagctaagta tgcaaggtag ttccagaaac atatctcaag atatgacaca gacatcaggt 2221 acaaatctta cttcagaaga gcttcggaag agacgagaag cctactttga aaaacagcag 2281 caaaagcagc aacagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag 2341 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag 2401 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag 2461 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag 2521 cagcagcggg acctatcagg acagagttca catccatgtg aaaggccagc caccagttca 2581 ggagcacttg ggagtgatct aggtaaggcc tgctcaccat tcatcatgtt cgctaccttc 2641 acactttatc tgacataaga gcggatccac cggatctaga taactgatca taatcagcca 2701 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct 2761 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta 2821 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 2881 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt 2941 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa 3001 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag 3061 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg 3121 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga 3181 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg 3241 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg 3301 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc 3361 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt 3421 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc 3481 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg 3541 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 3601 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 3661 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 3721 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 3781 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt 3841 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca 3901 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg 3961 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag 4021 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc 4081 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc 4141 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg 4201 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc 4261 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca 4321 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag 4381 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg 4441 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg 4501 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca 4561 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc 4621 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg 4681 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct 4741 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt 4801 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc 4861 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc 4921 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg 4981 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca 5041 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg 5101 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact 5161 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga 5221 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 5281 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 5341 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 5401 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta 5461 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 5521 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 5581 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 5641 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 5701 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 5761 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 5821 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 5881 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 5941 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat 6001 gcat //
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