质粒名称: | pmScarlet-i_C1 |
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出品公司: | Addgene |
目录编号: | 85044 |
存储实验室: | Dorus Gadella |
载体骨架基本信息 |
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载体名称: | pC1 |
载体出品公司: | Clontech |
载体抗性: | 卡那霉素 |
空载体大小: | 4000 bp |
完整质粒大小: | -- |
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: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | mScarlet-i |
物种来源: | 合成的 |
插入大小: | 701 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | CMV |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | AgeI(未破坏) |
3' 克隆位点: | BsrGI(未破坏) |
5' 测序引物 | CMV_fwd_primer |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pmScarlet-i_C1 was a gift from Dorus Gadella (Addgene plasmid # 85044 ; http://n2t.net/addgene:85044 ; RRID:Addgene_85044) |
参考文献部分: | mScarlet: a bright monomeric red fluorescent protein for cellular imaging. Bindels DS, Haarbosch L, van Weeren L, Postma M, Wiese KE, Mastop M, Aumonier S, Gotthard G, Royant A, Hink MA, Gadella TW Jr. Nat Methods. 2016 Nov 21. doi: 10.1038/nmeth.4074. 10.1038/nmeth.4074 PubMed 27869816 |
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pmScarlet-i_C1载体基本信息
pmScarlet-i_C1载体图谱质粒图谱和多克隆位点信息
pmScarlet-i_C1载体简介
pmScarlet-i_C1载体序列
LOCUS pmScarlet-i_C1 4710 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Mammalian expression of mScarlet-i. ACCESSION . VERSION . KEYWORDS pmScarlet-i_C1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4710) AUTHORS Bindels DS, Haarbosch L, van Weeren L, Postma M, Wiese KE, Mastop M, Aumonier S, Gotthard G, Royant A, Hink MA, Gadella TW Jr TITLE mScarlet: a bright monomeric red fluorescent protein for cellular imaging. JOURNAL Nat Methods. 2016 Nov 21. doi: 10.1038/nmeth.4074. PUBMED 27869816 REFERENCE 2 (bases 1 to 4710) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4710 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 119..422 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 423..626 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 577..597 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 665..674 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 671..1366 /codon_start=1 /product="bright monomeric red fluorescent protein evolved from a synthetic template (Bindels et al., 2016)" /label=mScarlet-I /note="matures faster than mScarlet" /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH STGGMDELYK" primer_bind complement(804..822) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1072..1091 /label=mCherry-F /note="mCherry, forward primer" misc_feature 1367..1432 /label=MCS /note="multiple cloning site" polyA_signal 1556..1677 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1593..1612) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1647..1666 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1684..2139) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1821..1842) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2033..2052 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2166..2270 /gene="bla" /label=AmpR promoter promoter 2272..2629 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2295..2315) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2480..2615 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2542..2561 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2664..3458 /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(2718..2737) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3328..3347 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3646..3665) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3690..3737 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4066..4654 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4555..4574 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta 61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc 661 ggtcgccacc atggtgagca agggcgaggc agtgatcaag gagttcatgc ggttcaaggt 721 gcacatggag ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg 781 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt 841 ctcctgggac atcctgtccc ctcagttcat gtacggctcc agggccttca tcaagcaccc 901 cgccgacatc cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt 961 gatgaacttc gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg 1021 caccctgatc tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat 1081 gcagaagaag acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt 1141 gctgaagggc gacattaaga tggccctgcg cctgaaggac ggcggccgct acctggcgga 1201 cttcaagacc acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga 1261 ccgcaagttg gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg 1321 ctccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtccg gactcagatc 1381 tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc 1441 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa 1501 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt 1561 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 1621 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg 1681 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct 1741 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg 1801 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact 1861 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac 1921 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga 1981 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga 2041 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca 2101 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa 2161 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca 2221 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg 2281 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc 2341 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc 2401 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt 2461 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc 2521 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct 2581 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag 2641 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 2701 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 2761 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 2821 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga 2881 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 2941 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 3001 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 3061 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 3121 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 3181 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 3241 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 3301 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 3361 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 3421 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat 3481 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta 3541 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg 3601 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga 3661 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg 3721 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac 3781 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc 3841 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat 3901 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag 3961 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc 4021 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt 4081 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt 4141 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat 4201 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc 4261 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa 4321 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg 4381 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag 4441 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag 4501 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa 4561 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt 4621 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg 4681 gttcctggcc ttttgctggc cttttgctca //
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