質粒名稱: | CapChR2_pmScarlet-N1 |
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出品公司: | Addgene |
目錄編號: | 188032 |
存儲實驗室: | Peter Hegemann |
載體骨架基本信息 |
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載體名稱: | pN1 |
載體出品公司: | Clontech |
空載體大小: | 4000bp |
完整質粒大小: | 5581bp |
載體類型: | 哺乳細胞表達 |
載體抗性: | Kanamycin, 50 μg/mL |
生長溫度: | 37 ℃ |
宿主菌: | DH5a |
拷貝數: | 未知 |
插入基因信息 |
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插入基因名稱: | CapChR2-mScarlet、Calcium permeable Channelrhodopsin 2 |
物種來源: | Chloromonas oogama |
插入大小: | 1578bp |
基因突變: | S43D, L112C, T139C and N238E mutations in CoChR |
啟動子: | CMV |
標簽/融合蛋白: | mScarlet (C terminal on insert) |
克隆方法: | Gibson Cloning |
5’測序引物 | CMV-F |
3’測序引物 | EGFP-C1-R |
文獻引用方法 |
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材料和方法部分: | CapChR2_pmScarlet-N1 was a gift from Peter Hegemann (Addgene plasmid # 188032 ; //n2t.net/addgene:188032 ; RRID:Addgene_188032) |
參考文獻部分: | Calcium-permeable channelrhodopsins for the photocontrol of calcium signalling. Fernandez Lahore RG, Pampaloni NP, Peter E, Heim MM, Tillert L, Vierock J, Oppermann J, Walther J, Schmitz D, Owald D, Plested AJR, Rost BR, Hegemann P. Nat Commun. 2022 Dec 21;13(1):7844. doi: 10.1038/s41467-022-35373-4. 10.1038/s41467-022-35373-4 PubMed 36543773 |
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CapChR2_pmScarlet-N1載體質粒基本信息
CapChR2_pmScarlet-N1質粒圖譜載體圖譜和CapChR2_pmScarlet-N1載體序列質粒序列多克隆位點信息
CapChR2_pmScarlet-N1質粒載體簡介
CapChR2_pmScarlet-N1質粒序列載體序列
LOCUS CapChR2_pmScarlet-N1 5581 bp DNA circular SYN 14-APR-2025 DEFINITION Expresses CapChR2-mScarlet in mammalian cells. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5581) AUTHORS Fernandez Lahore RG, Pampaloni NP, Peter E, Heim MM, Tillert L, Vierock J, Oppermann J, Walther J, Schmitz D, Owald D, Plested AJR, Rost BR, Hegemann P TITLE Calcium-permeable channelrhodopsins for the photocontrol of calcium signalling. JOURNAL Nat Commun. 2022 Dec 21;13(1):7844. doi: 10.1038/s41467-022-35373-4. PUBMED 36543773 REFERENCE 2 (bases 1 to 5581) 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: "Nat Commun."; date: "2022-12-21"; volume: "13(1):7844. doi"; pages: " 10.1038/s41467-022-35373-4" COMMENT Sequence Label: CapChR2_pmScarlet-N1 FEATURES Location/Qualifiers source 1..5581 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 153..456 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 457..660 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 611..631 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 697..1560 /codon_start=1 /product="high-photocurrent channelrhodopsin from Chloromonas oogama (Shemesh et al., 2017)" /label=CoChR /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFDIL ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW LLTCPVICIHLSNLTGLKDDYSKRTMRLLVSDVGCIVWGATSAMSTGYVKVIFFVLGCI YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV YGSTIGHTIIDLMSKECWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD QEDEETV" CDS 1579..2271 /codon_start=1 /product="bright monomeric red fluorescent protein evolved from a synthetic template (Bindels et al., 2016)" /label=mScarlet /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS TGGMDELYK" primer_bind complement(1709..1727) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1977..1996 /label=mCherry-F /note="mCherry, forward primer" misc_feature 2281..2337 /label=MCS /note="multiple cloning site" polyA_signal 2461..2582 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2498..2517) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2552..2571 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2589..3044) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2726..2747) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2938..2957 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3071..3175 /gene="bla" /label=AmpR promoter promoter 3177..3534 /label=SV40 promoter /note="SV40 enhancer and early promoter" enhancer 3177..3368 /label=SV40 enhancer /note="enhancer for the SV40 early promoter (Herr, 1993)" primer_bind complement(3200..3220) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3385..3520 /label=SV40 ori /note="SV40 origin of replication" CDS 3569..4363 /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 3569..4363 /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(3623..3642) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4233..4252 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4551..4570) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4595..4642 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4971..5559 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5460..5479 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 61 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg 121 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg 181 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata 241 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc 301 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac 361 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg 421 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc 481 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc 541 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc 601 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct 661 ggtttagtga accgtcagat ccgctagcgc taccggatgc tgggaaacgg cagcgccatt 721 gtgcctatcg accagtgctt ttgcctggct tggaccgaca gcctgggaag cgatacagag 781 cagctggtgg ccaacatcct ccagtggttc gccttcggct tcgacatcct gatcctgatg 841 ttctacgcct accagacttg gagagccact tgcggttggg aggaggtcta cgtctgttgc 901 gtcgagctga ccaaggtcat catcgagttc ttccacgagt tcgacgaccc cagcatgctg 961 tacctggcta acggacaccg agtccagtgg ctgagatacg cagagtggct gctgacttgt 1021 cccgtcatct gcatccacct gagcaacctg accggcctga aggacgacta cagcaagcgg 1081 accatgaggc tgctggtgtc agacgtggga tgcatcgtgt ggggagctac aagcgccatg 1141 agcacaggct acgtcaaggt catcttcttc gtgctgggtt gcatctacgg cgccaacacc 1201 ttcttccacg ccgccaaggt gtatatcgag agctaccacg tggtgccaaa gggcagacct 1261 agaaccgtcg tgcggatcat ggcttggctg ttcttcctgt cttggggcat gttccccgtg 1321 ctgttcgtcg tgggaccaga aggattcgac gccatcagcg tgtacggctc taccattggc 1381 cacaccatca tcgacctcat gagcaaggag tgttggggcc tgctgggaca ctatctgaga 1441 gtgctgatcc accagcacat catcatctac ggcgacatcc ggaagaagac caagatcaac 1501 gtggccggcg aggagatgga agtggagacc atggtggacc aggaggacga ggagacagtg 1561 gcggcaccgg tagtagcagt gagcaagggc gaggcagtga tcaaggagtt catgcggttc 1621 aaggtgcaca tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag 1681 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg 1741 cccttctcct gggacatcct gtcccctcag ttcatgtacg gctccagggc cttcaccaag 1801 caccccgccg acatccccga ctactataag cagtccttcc ccgagggctt caagtgggag 1861 cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga cccaggacac ctccctggag 1921 gacggcaccc tgatctacaa ggtgaagctc cgcggcacca acttccctcc tgacggcccc 1981 gtaatgcaga agaagacaat gggctgggaa gcgtccaccg agcggttgta ccccgaggac 2041 ggcgtgctga agggcgacat taagatggcc ctgcgcctga aggacggcgg ccgctacctg 2101 gcggacttca agaccaccta caaggccaag aagcccgtgc agatgcccgg cgcctacaac 2161 gtcgaccgca agttggacat cacctcccac aacgaggact acaccgtggt ggaacagtac 2221 gaacgctccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaaggactc 2281 agatctcgag ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccacc 2341 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta 2401 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt 2461 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 2521 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 2581 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat 2641 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata 2701 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt 2761 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc 2821 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa 2881 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg 2941 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt 3001 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg 3061 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc 3121 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga 3181 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc 3241 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag 3301 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 3361 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 3421 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 3481 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga 3541 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc 3601 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg 3661 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac 3721 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc 3781 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg 3841 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga 3901 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg 3961 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg 4021 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt 4081 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc 4141 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg 4201 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga 4261 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc 4321 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc 4381 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc 4441 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 4501 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg 4561 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg 4621 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc 4681 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc 4741 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct 4801 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt 4861 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 4921 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 4981 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 5041 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 5101 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 5161 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 5221 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 5281 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 5341 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 5401 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 5461 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 5521 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 5581 t //
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