質粒名稱: | mCherry-Sec61b-C1 |
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出品公司: | Addgene |
目錄編號: | 90994 |
存儲實驗室: | Jennifer Lippincott-Schwartz |
載體骨架基本信息 |
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載體名稱: | pEGFP-C1 |
載體出品公司: | Clontech |
空載體大小: | 4732bp |
完整質粒大小: | 5000bp |
載體類型: | 哺乳細胞表達 |
篩選標記: | 新霉素(G418) |
載體抗性: | Kanamycin, 50 μg/mL |
生長溫度: | 37 ℃ |
宿主菌: | DH5a |
拷貝數: | 高拷貝 |
插入基因信息 |
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插入基因名稱: | mCherry-Sec61b |
物種來源: | H. sapiens (human), Synthetic |
插入大小: | 702bp |
啟動子: | CMV |
標簽/融合蛋白: | mCherry (red fluorescence) (N terminal on insert) |
克隆方法: | 限制性內切酶 |
5’克隆位點: | AgeI (not destroyed) |
3’克隆位點: | BsrG1 (not destroyed) |
5’測序引物 | CMV-F |
3’測序引物 | SV40polyA |
文獻引用方法 |
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材料和方法部分: | mCherry-Sec61b-C1 was a gift from Jennifer Lippincott-Schwartz (Addgene plasmid # 90994 ; //n2t.net/addgene:90994 ; RRID:Addgene_90994) |
參考文獻部分: | Increased spatiotemporal resolution reveals highly dynamic dense tubular matrices in the peripheral ER. Nixon-Abell J, Obara CJ, Weigel AV, Li D, Legant WR, Xu CS, Pasolli HA, Harvey K, Hess HF, Betzig E, Blackstone C, Lippincott-Schwartz J. Science. 2016 Oct 28;354(6311). pii: aaf3928. Epub 2016 Oct 27. 10.1126/science.aaf3928 PubMed 27789813 |
買家導航
mCherry-Sec61b-C1載體質粒基本信息
mCherry-Sec61b-C1質粒圖譜載體圖譜和 mCherry-Sec61b-C1載體序列質粒序列多克隆位點信息
mCherry-Sec61b-C1質粒載體簡介
mCherry-Sec61b-C1質粒序列載體序列
LOCUS mCherry-Sec61b-C1 4999 bp DNA circular SYN 14-APR-2025 DEFINITION Expresses mCherry-tagged Sec61b, brightly labels endoplasmic reticulum in mammalian cells. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4999) AUTHORS Nixon-Abell J, Obara CJ, Weigel AV, Li D, Legant WR, Xu CS, Pasolli HA, Harvey K, Hess HF, Betzig E, Blackstone C, Lippincott-Schwartz J TITLE Increased spatiotemporal resolution reveals highly dynamic dense tubular matrices in the peripheral ER. JOURNAL Science. 2016 Oct 28;354(6311). pii: aaf3928. Epub 2016 Oct 27. PUBMED 27789813 REFERENCE 2 (bases 1 to 4999) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1 //gt12365.cn COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Science."; date: "2016-10-28"; volume: "354(6311). pii"; pages: " aaf3928. Epub 2016 Oct 27" COMMENT Sequence Label: mCherry-Sec61b-C1 FEATURES Location/Qualifiers source 1..4999 /mol_type="other DNA" /organism="synthetic DNA construct" polyA_signal 403..450 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" primer_bind 475..494 /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" CDS complement(682..1476) /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 complement(682..1476) /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(793..812) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 1403..1422 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter complement(1511..1868) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 1525..1660 /label=SV40 ori /note="SV40 origin of replication" enhancer 1677..1868 /label=SV40 enhancer /note="enhancer for the SV40 early promoter (Herr, 1993)" primer_bind 1825..1845 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter complement(1870..1974) /gene="bla" /label=AmpR promoter rep_origin 2001..2456 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2088..2107) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2298..2319 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 2463..2584 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2474..2493) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2528..2547 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(2745..3035) /codon_start=1 /gene="SEC61B" /product="human Sec61-beta subunit of the translocon in the endoplasmic reticulum" /label=Sec61-beta /translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS" CDS complement(3051..3758) /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(3326..3345) /label=mCherry-F /note="mCherry, forward primer" primer_bind 3595..3613 /label=mCherry-R /note="mCherry, reverse primer" regulatory 3755..3764 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" promoter complement(3803..4006) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(3832..3852) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 4007..4310 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" rep_origin complement(join(4485..4999,1..74)) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4565..4584) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 61 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 121 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 181 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta 241 tggcagggcc tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg 301 ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg 361 tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac 421 cgtgcgtttt attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct 481 ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc 541 cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat 601 agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca 661 tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg 721 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc 781 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc 841 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa 901 gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct 961 ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac 1021 aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa 1081 tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac 1141 tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag 1201 cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt 1261 cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag 1321 gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc 1381 agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc 1441 cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc 1501 ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata 1561 gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg 1621 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga 1681 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg 1741 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg 1801 ctggggagcc tggggacttt ccacacccta actgacacac attccacagc tggttctttc 1861 cgcctcagga ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 1921 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 1981 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 2041 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 2101 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 2161 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 2221 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 2281 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 2341 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 2401 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttacgc 2461 gttaagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt 2521 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca 2581 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt 2641 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agttatctag 2701 atccggtgga tcccgggccc gcggtaccgt cgactgcaga attcctacga acgagtgtac 2761 ttgccccaaa tgtgcaacat aaatacagaa gcgatgaaca gaagactcat aaccaatact 2821 ggaacagggc caactttgag cccaggtgaa tcttctgtgt agaatcgcca catccccccg 2881 gtgcctgccg aggttgtgcg gcctgcactc cttgtcccac agctggcatt tttcctctgc 2941 cggacagtgg atcccgccgc ccgggcggcc actgctttgc tgggagagcg ccctgaggat 3001 cccacgttag tgccactggg ggtcggacca ggcatagatc tgagtccgga cttgtacagc 3061 tcgtccatgc cgccggtgga gtggcggccc tcggcgcgtt cgtactgttc cacgatggtg 3121 tagtcctcgt tgtgggaggt gatgtccaac ttgatgttga cgttgtaggc gccgggcagc 3181 tgcacgggct tcttggcctt gtaggtggtc ttgacctcag cgtcgtagtg gccgccgtcc 3241 ttcagcttca gcctctgctt gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc 3301 tcggaggagg cctcccagcc catggtcttc ttctgcatta cggggccgtc ggaggggaag 3361 ttggtgccgc gcagcttcac cttgtagatg aactcgccgt cctgcaggga ggagtcctgg 3421 gtcacggtca ccacgccgcc gtcctcgaag ttcatcacgc gctcccactt gaagccctcg 3481 gggaaggaca gcttcaagta gtcggggatg tcggcggggt gcttcacgta ggccttggag 3541 ccgtacatga actgagggga caggatgtcc caggcgaagg gcagggggcc acccttggtc 3601 accttcagct tggcggtctg ggtgccctcg taggggcggc cctcgccctc gccctcgatc 3661 tcgaactcgt ggccgttcac ggagccctcc atgtgcacct tgaagcgcat gaactccttg 3721 atgatggcca tgttatcctc ctcgcccttg ctcaccatgg tggcgaccgg tagcgctagc 3781 ggatctgacg gttcactaaa ccagctctgc ttatatagac ctcccaccgt acacgcctac 3841 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt 3901 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt 3961 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga 4021 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat 4081 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat 4141 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat 4201 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg 4261 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca 4321 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta atgcatggcg 4381 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 4441 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 4501 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 4561 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 4621 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 4681 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 4741 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 4801 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 4861 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 4921 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 4981 ggtagctctt gatccggca //
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