| 质粒名称: | CapChR2_pmScarlet-N1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 188032 |
| 存储实验室: | Peter Hegemann |
|
载体骨架基本信息 |
|
| 载体名称: | pN1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4000bp |
| 完整质粒大小: | 5581bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | Kanamycin, 50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | CapChR2-mScarlet、Calcium permeable Channelrhodopsin 2 |
| 物种来源: | Chloromonas oogama |
| 插入大小: | 1578bp |
| 基因突变: | S43D, L112C, T139C and N238E mutations in CoChR |
| 启动子: | CMV |
| 标签/融合蛋白: | mScarlet (C terminal on insert) |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | CMV-F |
| 3’测序引物 | EGFP-C1-R |
|
文献引用方法 |
|
| 材料和方法部分: | CapChR2_pmScarlet-N1 was a gift from Peter Hegemann (Addgene plasmid # 188032 ; http://n2t.net/addgene:188032 ; RRID:Addgene_188032) |
| 参考文献部分: | 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 |
买家导航
CapChR2_pmScarlet-N1载体质粒基本信息
CapChR2_pmScarlet-N1质粒图谱载体图谱和CapChR2_pmScarlet-N1载体序列质粒序列多克隆位点信息
CapChR2_pmScarlet-N1质粒载体简介
CapChR2_pmScarlet-N1质粒序列载体序列
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
https://www.biofeng.com
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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