质粒名称: | pmTurquoise2-Golgi |
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出品公司: | Addgene |
目录编号: | 36205 |
存储实验室: | Dorus Gadella |
载体骨架基本信息 |
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载体名称: | pmTurquoise2-N1 |
载体出品公司: | Clontech |
空载体大小: | 4904bp |
完整质粒大小: | 4904bp |
载体类型: | 哺乳细胞表达 |
筛选标记: | 新霉素(G418) |
载体抗性: | Kanamycin |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | Golgi targeting sequence / B4GALT1(1-61) /beta4Gal-T1(1-61)/ beta-1,4-galactosyltransferase 1(1-61) |
物种来源: | H. sapiens (human) |
插入大小: | 183bp |
GenBank编号 | NM_001497 |
启动子: | CMV |
标签/融合蛋白: | mTurquoise2 (C terminal on backbone) |
克隆方法: | 限制性内切酶 |
5’克隆位点: | BglII (not destroyed) |
3’克隆位点: | AgeI (not destroyed) |
5’测序引物 | AGGTCTATATAAGCAGAGC |
3’测序引物 | TCTACAAATGTGGTATGGC |
文献引用方法 |
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材料和方法部分: | pmTurquoise2-Golgi was a gift from Dorus Gadella (Addgene plasmid # 36205 ; http://n2t.net/addgene:36205 ; RRID:Addgene_36205) |
参考文献部分: | Structure-guided evolution of cyan fluorescent proteins towards a quantum yield of 93%. Goedhart J, von Stetten D, Noirclerc-Savoye M, Lelimousin M, Joosen L, Hink MA, van Weeren L, Gadella TW Jr, Royant A. Nat Commun. 2012 Mar 20;3:751. doi: 10.1038/ncomms1738. 10.1038/ncomms1738 PubMed 22434194 |
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载体基本信息
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载体简介
载体序列
LOCUS pmTurquoise2-Golgi 4904 bp ds-DNA circular SYN 17-JUN-2022 DEFINITION In vivo visualization of golgi apparatus (can be used for colocalization studies). ACCESSION . VERSION . KEYWORDS pmTurquoise2-Golgi SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4904) AUTHORS Goedhart J, von Stetten D, Noirclerc-Savoye M, Lelimousin M, Joosen L, Hink MA, van Weeren L, Gadella TW Jr, Royant A TITLE Structure-guided evolution of cyan fluorescent proteins towards a quantum yield of 93%. JOURNAL Nat Commun. 2012 Mar 20;3:751. doi: 10.1038/ncomms1738. PUBMED 22434194 REFERENCE 2 (bases 1 to 4904) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 17, 2022 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..4904 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 9..312 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 313..516 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 467..487 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 792..801 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 798..1517 /codon_start=1 /product="enhanced monomeric variant of CFP (Goedhart et al., 2012)" /label=mTurquoise2 /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(843..864) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1104..1123) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1451..1472 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1640..1761 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1677..1696) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1731..1750 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1768..2223) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1905..1926) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2117..2136 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2250..2354 /gene="bla" /label=AmpR promoter promoter 2356..2713 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2379..2399) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2564..2699 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2626..2645 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2748..3542 /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(2802..2821) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3412..3431 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3730..3749) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3774..3821 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4150..4738 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4639..4658 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 61 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 121 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 181 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 241 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 301 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac 361 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 421 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 481 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc 541 tagcgctacc ggactcagat ctcgagctcg ccaccatgag gcttcgggag ccgctcctga 601 gcggcagcgc cgcgatgcca ggcgcgtccc tacagcgggc ctgccgcctg ctcgtggccg 661 tctgcgctct gcaccttggc gtcaccctcg tttactacct ggctggccgc gacctgagcc 721 gcctgcccca actggtcgga gtctccacac cgctgcagtc gacggtaccg cgggcccggg 781 atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca 841 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg 901 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc 961 ccgtgccctg gcccaccctc gtgaccaccc tgtcctgggg cgtgcagtgc ttcgcccgct 1021 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc 1081 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt 1141 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg 1201 gcaacatcct ggggcacaag ctggagtaca actactttag cgacaacgtc tatatcaccg 1261 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg 1321 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc 1381 tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga 1441 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg 1501 acgagctgta caagtaaagc ggccgcgact ctagatcata atcagccata ccacatttgt 1561 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat 1621 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa 1681 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 1741 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg 1801 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc 1861 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga 1921 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg 1981 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag 2041 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga 2101 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg 2161 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg 2221 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 2281 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 2341 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg 2401 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 2461 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 2521 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc 2581 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc 2641 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct 2701 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag 2761 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg 2821 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc 2881 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag 2941 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca 3001 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat 3061 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata 3121 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac 3181 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc 3241 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg 3301 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg 3361 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta 3421 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg 3481 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct 3541 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga 3601 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc 3661 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg 3721 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat 3781 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc 3841 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt 3901 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa 3961 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt 4021 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac 4081 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 4141 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 4201 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 4261 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg 4321 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 4381 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 4441 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 4501 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 4561 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 4621 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 4681 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 4741 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt 4801 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt 4861 aatagtaatc aattacgggg tcattagttc atagcccata tatg //
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