| 质粒名称: | pEGFP-RhoA Biosensor |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 68026 |
| 存储实验室: | Michael Glotzer |
|
载体骨架基本信息 |
|
| 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: | 4700 bp |
| 完整质粒大小: | 6140 bp |
| 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: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Anillin (AHD+PH),RhoA Biosensor,AHPH,AHDPH |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1428 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 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: | HindIII(未破坏) |
| 3' 克隆位点: | SacII (未破坏) |
| 5' 测序引物 | CATGGACGAGCTGTACAAG |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pEGFP-RhoA Biosensor was a gift from Michael Glotzer (Addgene plasmid # 68026 ; http://n2t.net/addgene:68026 ; RRID:Addgene_68026) |
| 参考文献部分: | Anillin is a scaffold protein that links RhoA, actin, and myosin during cytokinesis. Piekny AJ, Glotzer M. Curr Biol. 2008 Jan 8;18(1):30-6. Epub 2007 Dec 27. 10.1016/j.cub.2007.11.068 PubMed 18158243 |
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LOCUS pEGFP-RhoA Biosensor 6140 bp ds-DNA circular SYN 24-MAY-2019 DEFINITION C-terminus of Anillin in mammalian expression vector pEGFP-C1. ACCESSION . VERSION . KEYWORDS pEGFP-RhoA Biosensor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6140) AUTHORS Piekny AJ, Glotzer M TITLE Anillin is a scaffold protein that links RhoA, actin, and myosin during cytokinesis. JOURNAL Curr Biol. 2008 Jan 8;18(1):30-6. Epub 2007 Dec 27. PUBMED 18158243 REFERENCE 2 (bases 1 to 6140) AUTHORS . TITLE Direct Submission JOURNAL Exported May 24, 2019 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6140 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 68..371 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 372..575 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 526..546 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 614..623 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 620..1336 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(665..686) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(926..945) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1273..1294 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2935..3056 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2972..2991) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3026..3045 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3063..3518) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3200..3221) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3412..3431 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3545..3649 /gene="bla" /label=AmpR promoter promoter 3651..4008 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3674..3694) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3859..3994 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3921..3940 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4043..4837 /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(4097..4116) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4707..4726 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5025..5044) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5069..5116 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5445..6033 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5934..5953 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg 61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc 121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt 181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc 241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact 421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa 481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta 541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct 601 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc 661 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg 721 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct 781 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg 841 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt 901 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa 961 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga 1021 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat 1081 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga 1141 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt 1201 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga 1261 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat 1321 ggacgagctg tacaagtccg gactcagatc tcgagctcaa gcttcgagaa ctcgtgtccc 1381 tcgagctgaa tctggtgata gccttggttc tgaagatcgt gatcttcttt acagcattga 1441 tgcatataga tctcaaagat tcaaagaaac agaacgtcca tcaataaagc aggtgattgt 1501 tcggaaggaa gatgttactt caaaactgga tgaaaaaaat aatgcctttc cttgtcaagt 1561 taatatcaaa cagaaaatgc aggaactcaa taacgaaata aatatgcaac agacagtgat 1621 ctatcaagct agccaggctc ttaactgctg tgttgatgaa gaacatggaa aagggtccct 1681 agaagaagct gaagcagaaa gacttcttct aattgcaact gggaagagaa cacttttgat 1741 tgatgaattg aataaattga agaacgaagg acctcagagg aagaataagg ctagtcccca 1801 aagtgaattt atgccatcca aaggatcagt tactttgtca gaaatccgct tgcctctaaa 1861 agcagatttt gtctgcagta cggttcagaa accagatgca gcaaattact attacttaat 1921 tatactaaaa gcaggagctg aaaatatggt agccacacca ttagcaagta cttcaaactc 1981 tcttaacggt gatgctctga cattcactac tacatttact ctgcaagatg tatccaatga 2041 ctttgaaata aatattgaag tttacagctt ggtgcaaaag aaagatccct caggccttga 2101 taagaagaaa aaaacatcca agtccaaggc tattactcca aagcgactcc tcacatctat 2161 aaccacaaaa agcaacattc attcttcagt catggccagt ccaggaggtc ttagtgctgt 2221 gcgaaccagc aacttcgccc ttgttggatc ttacacatta tcattgtctt cagtaggaaa 2281 tactaagttt gttctggaca aggtcccctt tttatcttct ttggaaggtc atatttattt 2341 aaaaataaaa tgtcaagtga attccagtgt tgaagaaaga ggttttctaa ccatatttga 2401 agatgttagt ggttttggtg cctggcatcg aagatggtgt gttctttctg gaaactgtat 2461 atcttattgg acttatccag atgatgagaa acgcaagaat cccataggaa ggataaatct 2521 ggctaattgt accagtcgtc agatagaacc agccaacaga gaattttgtg caagacgcaa 2581 cacttttgaa ttaattactg tccgaccaca aagagaagat gaccgagaga ctcttgtcag 2641 ccaatgcagg gacacactct gtgttaccaa gaactggctg tctgcagata ctaaagaaga 2701 gcgggatctc tggatgcaaa aactcaatca agttcttgtt gatattcgcc tctggcaacc 2761 tgatgcttgc tacaaaccta ttggaaagcc ttgaccgcgg gcccgggatc caccggatct 2821 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac 2881 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg 2941 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 3001 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc 3061 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 3121 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 3181 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 3241 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 3301 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 3361 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 3421 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 3481 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa 3541 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 3601 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga 3661 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 3721 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 3781 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 3841 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 3901 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta 3961 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga 4021 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 4081 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 4141 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 4201 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac 4261 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 4321 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 4381 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 4441 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 4501 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 4561 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 4621 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 4681 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 4741 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 4801 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg 4861 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat 4921 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 4981 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag 5041 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt 5101 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc 5161 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc 5221 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata 5281 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg 5341 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg 5401 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt 5461 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 5521 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 5581 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 5641 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 5701 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 5761 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 5821 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 5881 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 5941 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 6001 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 6061 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct 6121 gtggataacc gtattaccgc //
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