| 质粒名称: | Grp1-PH pEGFP-C1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 71378 |
| 存储实验室: | Mark Lemmon |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-C1 |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 5178bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 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: | Grp1-PH |
| 物种来源: | M. musculus (mouse) |
| 基因突变: | TPleckstrin homology (PH) |
| 启动子: | CMV |
| 标签/融合蛋白: | EGFP (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BglII (unknown if destroyed) |
| 3’克隆位点: | EcoRI (not destroyed) |
| 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: | Grp1-PH pEGFP-C1 was a gift from Mark Lemmon (Addgene plasmid # 71378 ; http://n2t.net/addgene:71378 ; RRID:Addgene_71378) |
| 参考文献部分: | Specificity and promiscuity in phosphoinositide binding by pleckstrin homology domains. Kavran JM, Klein DE, Lee A, Falasca M, Isakoff SJ, Skolnik EY, Lemmon MA. J Biol Chem. 1998 Nov 13;273(46):30497-508. PubMed 9804818 |
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Grp1-PH pEGFP-C1载体质粒基本信息
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Grp1-PH pEGFP-C1质粒载体简介
Grp1-PH pEGFP-C1质粒序列载体序列
LOCUS Grp1-PH pEGFP-C1 5178 bp DNA circular SYN 14-APR-2025 DEFINITION For analysis of PH domain localization in mammalian cells. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5178) AUTHORS Kavran JM, Klein DE, Lee A, Falasca M, Isakoff SJ, Skolnik EY, Lemmon MA TITLE Specificity and promiscuity in phosphoinositide binding by pleckstrin homology domains. JOURNAL J Biol Chem. 1998 Nov 13;273(46):30497-508. PUBMED 9804818 REFERENCE 2 (bases 1 to 5178) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1 https://www.biofeng.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol Chem."; date: "1998-11-13"; volume: "273(46)"; pages: "30497-508" COMMENT Sequence Label: Grp1-PH pEGFP-C1 FEATURES Location/Qualifiers source 1..5178 /mol_type="other DNA" /organism="synthetic DNA construct" 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 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS join(613..615,616..618,619..1329) /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS join(613..615,616..618,619..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" polyA_signal 1966..2087 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2003..2022) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2057..2076 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2094..2549) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2231..2252) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2443..2462 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2576..2680 /gene="bla" /label=AmpR promoter promoter 2682..3039 /label=SV40 promoter /note="SV40 enhancer and early promoter" enhancer 2682..2873 /label=SV40 enhancer /note="enhancer for the SV40 early promoter (Herr, 1993)" primer_bind complement(2705..2725) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2890..3025 /label=SV40 ori /note="SV40 origin of replication" CDS 3074..3868 /codon_start=1 /product="aminoglycoside phosphotransferase" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" CDS 3074..3868 /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(3128..3147) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3738..3757 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4056..4075) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4100..4147 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4476..5064 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4965..4984 /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 atccccgttt aagatcccag aagacgacgg aaatgacctg 1381 acacacacgt tcttcaaccc agaccgagaa ggctggctgc tgaagctggg gggtcgtgtg 1441 aagacctgga aacggcgctg gttcatcctc acagataact gcctctacta ctttgagtac 1501 accacggaca aggagcccag gggcatcatc cccctggaga acctcagcat cagggaggtg 1561 gaggaccccc ggaagccgaa ctgctttgag ctgtataacc ccagtcacaa aggtcaagtc 1621 atcaaggcct gcaagacgga ggccgatggc cgtgtggtgg agggcaacca cgttgtgtac 1681 cggatctctg cccccagccc ggaggaaaag gaggagtgga tgaagtccat caaagcaagc 1741 atcagtaggg acccgttcta tgacatgttg gccacgagga aaaggaggat tgccaataag 1801 aaatagaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg 1861 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac 1921 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca 1981 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 2041 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt 2101 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa 2161 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt 2221 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa 2281 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag 2341 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt 2401 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg 2461 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc 2521 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg 2581 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata 2641 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag 2701 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt 2761 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca 2821 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta 2881 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 2941 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag 3001 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg 3061 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt 3121 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt 3181 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc 3241 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc 3301 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga 3361 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 3421 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca 3481 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga 3541 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc 3601 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat 3661 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 3721 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg 3781 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt 3841 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa 3901 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 3961 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 4021 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg 4081 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg 4141 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga 4201 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg 4261 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt 4321 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg 4381 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga 4441 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 4501 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 4561 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 4621 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 4681 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 4741 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 4801 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 4861 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 4921 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 4981 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 5041 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 5101 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac 5161 cgtattaccg ccatgcat //
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