质粒名称: | CRY2low-mCherry |
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出品公司: | addgene |
目录编号: | 104065 |
存储实验室: | Bianxiao Cui |
载体骨架基本信息 |
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载体名称: | pmCherryN1 |
空载体大小: | 4000 bp |
完整质粒大小: | 6200 bp |
载体类型: | 哺乳细胞表达 |
载体抗性: | Kanamycin |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 未知 |
插入基因信息 |
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插入基因名称: | CRY2low-mCherry/CRY2PHR/Cherry |
物种来源: | A. thaliana (mustard weed)芥花(芥菜杂草) |
插入大小: | 951 bp |
基因突变: | CRY2 aa 1-491 truncation, R489E A491D |
启动子: | CMV |
克隆方法: | Gibson Cloning |
5’测序引物 | CMV forward |
文献引用方法 |
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材料和方法部分: | CRY2low-mCherry was a gift from Bianxiao Cui (Addgene plasmid # 104065 ; http://n2t.net/addgene:104065 ; RRID:Addgene_104065) |
参考文献部分: | Understanding CRY2 interactions for optical control of intracellular signaling. Duan L, Hope J, Ong Q, Lou HY, Kim N, McCarthy C, Acero V, Lin MZ, Cui B. Nat Commun. 2017 Sep 15;8(1):547. doi: 10.1038/s41467-017-00648-8. 10.1038/s41467-017-00648-8 [pii] PubMed 28916751 |
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载体序列
LOCUS CRY2low-mCherry 6167 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses fusion of CRY2low mutant (CRY2 aa1-491 R489E A491D) with mCherry. ACCESSION . VERSION . KEYWORDS CRY2low-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6167) AUTHORS Duan L, Hope J, Ong Q, Lou HY, Kim N, McCarthy C, Acero V, Lin MZ, Cui B TITLE Understanding CRY2 interactions for optical control of intracellular signaling. JOURNAL Nat Commun. 2017 Sep 15;8(1):547. doi: 10.1038/s41467-017-00648-8. PUBMED 28916751 REFERENCE 2 (bases 1 to 6167) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..6167 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 73..376 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 377..580 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 531..551 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 2128..2137 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2134..2844 /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(2279..2297) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2547..2566 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2967..3088 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3004..3023) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3058..3077 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3095..3550) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3232..3253) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3444..3463 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3577..3681 /gene="bla" /label=AmpR promoter promoter 3683..4040 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3706..3726) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3891..4026 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3953..3972 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4075..4869 /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(4129..4148) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4739..4758 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5057..5076) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5101..5148 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5477..6065 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5966..5985 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata 61 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 121 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 181 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 241 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 301 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 361 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 421 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 481 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 541 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat 601 ccgctagcgc taccggactc agatctcgag gccaccatga agatggacaa aaagactata 661 gtttggttta gaagagacct aaggattgag gataatcctg cattagcagc agctgctcac 721 gaaggatctg tttttcctgt cttcatttgg tgtcctgaag aagaaggaca gttttatcct 781 ggaagagctt caagatggtg gatgaaacaa tcacttgctc acttatctca atccttgaag 841 gctcttggat ctgacctcac tttaatcaaa acccacaaca cgatttcagc gatcttggat 901 tgtatccgcg ttaccggtgc tacaaaagtc gtctttaacc acctctatga tcctgtttcg 961 ttagttcggg accataccgt aaaggagaag ctggtggaac gtgggatctc tgtgcaaagc 1021 tacaatggag atctattgta tgaaccgtgg gagatatact gcgaaaaggg caaacctttt 1081 acgagtttca attcttactg gaagaaatgc ttagatatgt cgattgaatc cgttatgctt 1141 cctcctcctt ggcggttgat gccaataact gcagcggctg aagcgatttg ggcgtgttcg 1201 attgaagaac tagggctgga gaatgaggcc gagaaaccga gcaatgcgtt gttaactaga 1261 gcttggtctc caggatggag caatgctgat aagttactaa atgagttcat cgagaagcag 1321 ttgatagatt atgcaaagaa cagcaagaaa gttgttggga attctacttc actactttct 1381 ccgtatctcc atttcgggga aataagcgtc agacacgttt tccagtgtgc ccggatgaaa 1441 caaattatat gggcaagaga taagaacagt gaaggagaag aaagtgcaga tctttttctt 1501 aggggaatcg gtttaagaga gtattctcgg tatatatgtt tcaacttccc gtttactcac 1561 gagcaatcgt tgttgagtca tcttcggttt ttcccttggg atgctgatgt tgataagttc 1621 aaggcctgga gacaaggcag gaccggttat ccgttggtgg atgccggaat gagagagctt 1681 tgggctaccg gatggatgca taacagaata agagtgattg tttcaagctt tgctgtgaag 1741 tttcttctcc ttccatggaa atggggaatg aagtatttct gggatacact tttggatgct 1801 gatttggaat gtgacatcct tggctggcag tatatctctg ggagtatccc cgatggccac 1861 gagcttgatc gcttggacaa tcccgcgtta caaggcgcca aatatgaccc agaaggtgag 1921 tacataaggc aatggcttcc cgagcttgcg agattgccaa ctgaatggat ccatcatcca 1981 tgggacgctc ctttaaccgt actcaaagct tctggtgtgg aactcggaac aaactatgcg 2041 aaacccattg tagacatcga cacagctcgt gagctactag ctaaagctat ttcaagaacc 2101 gaggaagacg cccgggatcc accggtcgcc accatggtga gcaagggcga ggaggataac 2161 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc 2221 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc 2281 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag 2341 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag 2401 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg 2461 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg 2521 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag 2581 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg 2641 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag 2701 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac 2761 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc 2821 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca 2881 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac 2941 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat 3001 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 3061 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta 3121 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc 3181 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg 3241 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat 3301 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc 3361 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag 3421 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg 3481 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta 3541 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt 3601 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa 3661 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt 3721 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3781 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3841 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 3901 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 3961 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg 4021 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt 4081 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 4141 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 4201 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac 4261 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 4321 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 4381 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 4441 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 4501 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 4561 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag 4621 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 4681 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 4741 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 4801 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 4861 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 4921 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 4981 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 5041 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga 5101 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg 5161 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg 5221 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg 5281 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga 5341 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc 5401 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 5461 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 5521 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 5581 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt 5641 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 5701 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 5761 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 5821 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 5881 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 5941 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 6001 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 6061 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 6121 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtatt //
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