质粒名称: | Beta-arrestin2 GFP WT |
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出品公司: | Addgene |
目录编号: | 35411 |
存储实验室: | Robert Lefkowitz |
载体骨架基本信息 |
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载体名称: | pEGFP-N1 |
载体出品公司: | Clontech |
载体抗性: | 卡那霉素 |
空载体大小: | 4700 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: | -- |
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: | βarrestin2 |
物种来源: | R. norvegicus (rat) |
插入大小: | 1500 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | CMV |
标签/融合蛋白: | EGFP(主干上的C端) |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | HindIII(未破坏) |
3' 克隆位点: | ApaI(未破坏) |
5' 测序引物 | -- |
3' 测序引物 | EGFP-N |
文献引用方法 |
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材料和方法部分: | Beta-arrestin2 GFP WT was a gift from Robert Lefkowitz (Addgene plasmid # 35411 ; http://n2t.net/addgene:35411 ; RRID:Addgene_35411) |
参考文献部分: | Receptor-specific ubiquitination of beta-arrestin directs assembly and targeting of seven-transmembrane receptor signalosomes. Shenoy SK, Lefkowitz RJ. J Biol Chem. 2005 Apr 15;280(15):15315-24. Epub 2005 Feb 7. 10.1074/jbc.M412418200 PubMed 15699045 |
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载体基本信息
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载体简介
载体序列
LOCUS Beta-arrestin2 GFP WT 5954 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Beta-arrestin2 GFP WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5954) AUTHORS Shenoy SK, Lefkowitz RJ TITLE Receptor-specific ubiquitination of beta-arrestin directs assembly and targeting of seven-transmembrane receptor signalosomes. JOURNAL J Biol Chem. 2005 Apr 15;280(15):15315-24. Epub 2005 Feb 7. PUBMED 15699045 REFERENCE 2 (bases 1 to 5954) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5954 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 305..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 1834..1843 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1840..2559 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1885..1906) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2146..2165) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2493..2514 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2682..2803 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2719..2738) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2773..2792 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2810..3265) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2947..2968) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3159..3178 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3292..3396 /gene="bla" /label=AmpR promoter promoter 3398..3755 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3421..3441) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3606..3741 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3668..3687 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3790..4584 /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(3844..3863) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4454..4473 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4772..4791) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4816..4863 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5192..5780 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5681..5700 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 541 ccggactcag atctcgagct caagcttggt accacgcgca ccatgggtga aaaacccggg 601 accagggtct tcaagaagtc gagccctaac tgcaagctca ccgtgtactt gggcaagcgt 661 gactttgtgg atcacttgga caaagtggat cctgtcgatg gtgtggtgct tgtggatcct 721 gactacttga aggaccggaa agtgtttgtg accctcacct gtgccttccg ctatggccga 781 gaagacctgg atgtactggg cctgtctttc cgcaaagatc tgttcatcgc cacctaccag 841 gccttccccc ccatgcccaa cccacctcgg ccccccaccc gcctacagga ccgactgctg 901 aagaagttgg gccagcacgc ccaccccttt tttttcacaa taccccagaa tttgccttgc 961 tccgtcacac tgcagccagg accggaggac acagggaagg cctgtggagt agactttgag 1021 attcgagcct tctgtgccaa atctatagaa gaaaaaagcc acaaaaggaa ctccgtgcgg 1081 cttatcatca gaaaggtaca gtttgctcct gagacacccg gcccccagcc atcagctgaa 1141 accacacgcc acttcctcat gtctgaccgg aggtccctgc acctagaggc ttccctggac 1201 aaagagctgt actaccatgg ggaacccctc aatgtcaacg tccacgtcac caacaattct 1261 gccaaaaccg tcaagaagat cagagtgtct gtgagacagt atgccgacat ttgcctcttc 1321 agcaccgcgc agtacaagtg tcctgtggct cagcttgaac aagatgacca ggtgtctccc 1381 agttccacat tctgcaaggt gtacaccata accccgctgc tcagtgacaa ccgagagaag 1441 cgtggccttg cccttgatgg gcaactcaag cacgaagaca ccaacctggc ttccagcacc 1501 attgtgaagg agggagccaa caaggaggtg ctgggaatcc tagtatccta cagggtcaag 1561 gtgaagctgg tggtgtctcg aggcggggat gtctccgtgg agctaccttt cgtcctaatg 1621 caccccaagc cccacgacca catcaccctt ccccgacccc agtcagcccc ccgggaaata 1681 gacatccctg tggataccaa cctcattgaa ttcgatacca actatgccac agacgacgac 1741 atcgtgtttg aggactttgc gaggcttcgg ctgaagggga tgaaggatga cgactgtgat 1801 gaccagttct gccgggcccg ggatccaccg gtcgccacca tggtgagcaa gggcgaggag 1861 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag 1921 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc 1981 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac 2041 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc 2101 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac 2161 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag 2221 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac 2281 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag 2341 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc 2401 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc 2461 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc 2521 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca 2581 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc 2641 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt 2701 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 2761 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc 2821 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa 2881 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt 2941 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg 3001 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt 3061 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga 3121 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg 3181 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg 3241 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc 3301 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc 3361 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt 3421 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 3481 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 3601 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 3661 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 3721 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga 3781 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 3841 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 3901 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 3961 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 4021 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 4081 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 4141 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 4201 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 4261 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc 4321 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 4381 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 4441 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 4501 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 4561 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga 4621 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 4681 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 4741 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag 4801 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg 4861 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc 4921 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg 4981 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact 5041 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 5101 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 5161 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 5221 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 5281 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc 5341 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 5401 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 5461 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 5521 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 5581 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 5641 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 5701 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 5761 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 5821 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta 5881 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca 5941 tatatggagt tccg //