| 质粒名称: | pEGFP-N1-Tspan4 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 210323 |
| 存储实验室: | Li Yu |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 5411bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | Kanamycin, 50 μg/mL |
| 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: | Tetraspanin 4 |
| 物种来源: | M. musculus (mouse) |
| 标签/融合蛋白: | GFP (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoR1 (unknown if destroyed) |
| 3’克隆位点: | Sal1 (unknown if destroyed) |
| 5’测序引物 | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | pEGFP-N1-Tspan4 was a gift from Li Yu (Addgene plasmid # 210323 ; http://n2t.net/addgene:210323 ; RRID:Addgene_210323) |
| 参考文献部分: | Discovery of the migrasome, an organelle mediating release of cytoplasmic contents during cell migration. Ma L, Li Y, Peng J, Wu D, Zhao X, Cui Y, Chen L, Yan X, Du Y, Yu L. Cell Res. 2015 Jan;25(1):24-38. doi: 10.1038/cr.2014.135. Epub 2014 Oct 24. 10.1038/cr.2014.135 PubMed 25342562 |
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pEGFP-N1-Tspan4载体质粒基本信息
pEGFP-N1-Tspan4质粒图谱载体图谱和pEGFP-N1-Tspan4载体序列质粒序列多克隆位点信息
pEGFP-N1-Tspan4质粒载体简介
pEGFP-N1-Tspan4质粒序列载体序列
LOCUS pEGFP-N1-Tspan4 5411 bp DNA circular SYN 14-APR-2025 DEFINITION Expresses Tetraspanin-4 (TSPAN4) as migrasome marker. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5411) AUTHORS Ma L, Li Y, Peng J, Wu D, Zhao X, Cui Y, Chen L, Yan X, Du Y, Yu L TITLE Discovery of the migrasome, an organelle mediating release of cytoplasmic contents during cell migration. JOURNAL Cell Res. 2015 Jan;25(1):24-38. doi: 10.1038/cr.2014.135. Epub 2014 Oct 24. PUBMED 25342562 REFERENCE 2 (bases 1 to 5411) 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"; doi: "10.1038/cr.2014.135"; journalName: "Cell Res"; date: "2015-01"; volume: "25"; issue: "1"; pages: "24-38" COMMENT Sequence Label: pEGFP-N1-Tspan4 FEATURES Location/Qualifiers source 1..5411 /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 1351..1360 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS join(1357..1359,1360..1362,1363..2076) /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS join(1357..1359,1360..1362,1363..2073) /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1402..1423) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1663..1682) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2010..2031 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2199..2320 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2236..2255) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2290..2309 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2327..2782) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2464..2485) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2676..2695 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2809..2913 /gene="bla" /label=AmpR promoter promoter 2915..3272 /label=SV40 promoter /note="SV40 enhancer and early promoter" enhancer 2915..3106 /label=SV40 enhancer /note="enhancer for the SV40 early promoter (Herr, 1993)" primer_bind complement(2938..2958) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3123..3258 /label=SV40 ori /note="SV40 origin of replication" CDS 3307..4101 /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 3307..4101 /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(3361..3380) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3971..3990 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4289..4308) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4333..4380 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4709..5297 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5198..5217 /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 ccggactcag atctcgagac catggcgcgc ggctgcctcc agggcgtcaa gtacctcatg 661 ttcgccttca acctgctctt ctggctgggt ggctgtggtg tcctgggtgt tggcatctgg 721 ttggctgcca cacagggaaa ctttgccacc ttatcatcct catttccatc cttgtcggct 781 gccaacctgc tcatcgtcac cgggaccttc gtcatggcca tcggcttcgt gggctgcatt 841 ggggccctca aggagaacaa gtgcctactg ctcactttct ttgtgctgct gctgctagtg 901 ttcctgctgg aagccaccat tgctgtgctc ttctttgcct acagtgacaa gattgacagt 961 tatgcccaac aagacctgaa gaagggcctg catctgtatg gcacacaggg caacgtgggc 1021 ctcaccaatg cctggagcat catccagact gatttccgat gctgtggagt ttccaattac 1081 actgattggt ttgaggtata caatgccact cgtgtgcctg actcctgctg tctggagttc 1141 agtgatagct gtgggttaca tgaacctggt acctggtgga agtcgccctg ttatgagaca 1201 gtgaaggcct ggctccagga gaacctgcta gctgtgggca tctttggact gtgcacggca 1261 ctggtgcaga ttctgggcct caccttcgct atgaccatgt actgccaggt ggtaaaggcg 1321 gacacctact gtgcacggga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg 1381 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc 1441 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc 1501 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc 1561 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc 1621 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag 1681 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc 1741 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc 1801 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc 1861 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc 1921 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg 1981 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc 2041 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa 2101 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc 2161 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata 2221 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 2281 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt 2341 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag 2401 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt 2461 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga 2521 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg 2581 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct 2641 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc 2701 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt 2761 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct 2821 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga 2881 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga 2941 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3001 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 3061 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 3121 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 3181 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag 3241 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg 3301 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg 3361 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg 3421 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat 3481 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca 3541 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg 3601 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat 3661 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa 3721 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg 3781 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg 3841 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg 3901 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat 3961 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac 4021 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc 4081 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc 4141 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg 4201 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt 4261 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa 4321 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc 4381 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt 4441 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag 4501 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat 4561 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc 4621 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 4681 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 4741 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 4801 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc 4861 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 4921 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 4981 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 5041 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 5101 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 5161 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 5221 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 5281 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct 5341 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta 5401 ccgccatgca t //
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