| 质粒名称: | NLS-mCherry-NES reporter (pDN160) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 72660 |
| 存储实验室: | Barbara Di Ventura, Roland Eils |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 (-) |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6250 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: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | NLS-mCherry-NES |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | NLS-mCherry-NES reporter (pDN160) was a gift from Barbara Di Ventura & Roland Eils (Addgene plasmid # 72660 ; http://n2t.net/addgene:72660 ; RRID:Addgene_72660) |
| 参考文献部分: | Optogenetic control of nuclear protein export. Niopek D, Wehler P, Roensch J, Eils R, Di Ventura B. Nat Commun. 2016 Feb 8;7:10624. doi: 10.1038/ncomms10624. 10.1038/ncomms10624 PubMed 26853913 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
质粒图谱.png)
载体简介
载体序列
LOCUS NLS-mCherry-NES reporter (pDN160) 6250 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION NLS-mCherry-NES 17 reporter for light-inducible nuclear export block
with pDN157 and pDN158.
ACCESSION .
VERSION .
KEYWORDS NLS-mCherry-NES reporter (pDN160)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6250)
AUTHORS Niopek D, Wehler P, Roensch J, Eils R, Di Ventura B
TITLE Optogenetic control of nuclear protein export.
JOURNAL Nat Commun. 2016 Feb 8;7:10624. doi: 10.1038/ncomms10624.
PUBMED 26853913
REFERENCE 2 (bases 1 to 6250)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6250
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 931..1635
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
GRHSTGGMDELYK"
primer_bind complement(1073..1091)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1341..1360
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind complement(1844..1861)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1850..2074
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2120..2548
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2207..2226)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2417..2438
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2557..2577)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2562..2891
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2742..2877
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2804..2823
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2958..3752
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3012..3031)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3622..3641
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3926..4047
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3963..3982)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4017..4036
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4096..4112)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4096..4112)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4109..4131)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4120..4136
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4144..4174)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4189..4210
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4327..4344)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4498..5083)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4578..4597)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5254..6114)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 5877..5896
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6115..6219)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 atggccgcag caaagcgcgt gaagctggac gtgagcaagg gcgaggagga taacatggcc
961 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
1021 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
1081 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
1141 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
1201 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
1261 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
1321 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
1381 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
1441 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
1501 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
1561 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
1621 gacgagctgt acaagggtgg atctggaggt tcaggtggaa gtgagggagt catgctgatt
1681 aagaaaactg cagaaaatat tgacgagctg gccaaggccc tgcccgacct gaacctggac
1741 tagctcgagc ggccgccact gtgctggata tctgcagaat tccaccacac tggactagtg
1801 gatccgagct cggtaccaag cttaagttta aaccgctgat cagcctcgac tgtgccttct
1861 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
1921 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
1981 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
2041 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
2101 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
2161 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
2221 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
2281 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
2341 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
2401 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
2461 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
2521 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
2581 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
2641 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
2701 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
2761 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
2821 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
2881 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
2941 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
3001 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
3061 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
3121 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
3181 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
3241 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
3301 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
3361 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
3421 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
3481 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
3541 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
3601 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
3661 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
3721 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
3781 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
3841 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
3901 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
3961 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
4021 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
4081 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
4141 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
4201 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
4261 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
4321 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
4381 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
4441 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
4501 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
4561 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
4621 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
4681 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
4741 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
4801 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
4861 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
4921 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
4981 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
5041 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
5101 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
5161 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
5221 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
5281 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
5341 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
5401 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
5461 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
5521 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
5581 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
5641 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
5701 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
5761 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
5821 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
5881 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
5941 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
6001 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
6061 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
6121 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
6181 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
6241 acctgacgtc
//
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