| 质粒名称: | pcDNA3-TLR3-CFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 13641 |
| 存储实验室: | Doug Golenbock |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3-CFP |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6100 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 |
|
| 插入基因名称: | Toll-Like Receptor 3,TLR3 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | CFP（主干上的C终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | HindIII(未破坏) |
| 3' 克隆位点: | XhoI(未破坏) |
| 5' 测序引物 | T7 |
| 3' 测序引物 | GTCTTGTAGTTGCCGTCGTC |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3-TLR3-CFP was a gift from Doug Golenbock (Addgene plasmid # 13641 ; http://n2t.net/addgene:13641 ; RRID:Addgene_13641) |
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
载体简介
载体序列
LOCUS pcDNA3-TLR3-CFP 8796 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3-TLR3-CFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8796)
TITLE TLR Signaling Plasmids
REFERENCE 2 (bases 1 to 8796)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8796
/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 3613..4332
/codon_start=1
/product="cyan variant of GFP"
/label=CFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3658..3679)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3919..3938)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4266..4287
/label=EGFP-C
/note="EGFP, forward primer"
promoter complement(4348..4366)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(4349..4366)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(4386..4403)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4392..4616
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 4662..5090
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4749..4768)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4959..4980
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5099..5119)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 5104..5434
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5285..5420
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5347..5366
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5501..6295
/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(5555..5574)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6165..6184
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 6469..6590
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6506..6525)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6560..6579
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6639..6655)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6639..6655)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6652..6674)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6663..6679
/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(6687..6717)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6732..6753
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6870..6887)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7041..7629)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7121..7140)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7800..8660)
/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 8423..8442
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8661..8765)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttatgaga
901 cagactttgc cttgtatcta cttttggggg ggcctcttgc cctttgggat gctgtgtgca
961 tcctccacca ccaagtgcac tgttagccat gaagttgctg actgcagcca cctgaagttg
1021 actcaggtac ccgatgatct acccacaaac ataacagtgt tgaaccttac ccataatcaa
1081 ctcagaagat taccagccgc caacttcaca aggtatagcc agctaactag cttggatgta
1141 ggatttaaca ccatctcaaa actggagcca gaattgtgcc agaaacttcc catgttaaaa
1201 gttttgaacc tccagcacaa tgagctatct caactttctg ataaaacctt tgccttctgc
1261 acgaatttga ctgaactcca tctcatgtcc aactcaatcc agaaaattaa aaataatccc
1321 tttgtcaagc agaagaattt aatcacatta gatctgtctc ataatggctt gtcatctaca
1381 aaattaggaa ctcaggttca gctggaaaat ctccaagagc ttctattatc aaacaataaa
1441 attcaagcgc taaaaagtga agaactggat atctttgcca attcatcttt aaaaaaatta
1501 gagttgtcat cgaatcaaat taaagagttt tctccagggt gttttcacgc aattggaaga
1561 ttatttggcc tctttctgaa caatgtccag ctgggtccca gccttacaga gaagctatgt
1621 ttggaattag caaacacaag cattcggaat ctgtctctga gtaacagcca gctgtccacc
1681 accagcaata caactttctt gggactaaag tggacaaatc tcactatgct cgatctttcc
1741 tacaacaact taaatgtggt tggtaacgat tcctttgctt ggcttccaca actagaatat
1801 ttcttcctag agtataataa tatacagcat ttgttttctc actctttgca cgggcttttc
1861 aatgtgaggt acctgaattt gaaacggtct tttactaaac aaagtatttc ccttgcctca
1921 ctccccaaga ttgatgattt ttcttttcag tggctaaaat gtttggagca ccttaacatg
1981 gaagataatg atattccagg cataaaaagc aatatgttca caggattgat aaacctgaaa
2041 tacttaagtc tatccaactc ctttacaagt ttgcgaactt tgacaaatga aacatttgta
2101 tcacttgctc attctccctt acacatactc aacctaacca agaataaaat ctcaaaaata
2161 gagagtgatg ctttctcttg gttgggccac ctagaagtac ttgacctggg ccttaatgaa
2221 attgggcaag aactcacagg ccaggaatgg agaggtctag aaaatatttt cgaaatctat
2281 ctttcctaca acaagtacct gcagctgact aggaactcct ttgccttggt cccaagcctt
2341 caacgactga tgctccgaag ggtggccctt aaaaatgtgg atagctctcc ttcaccattc
2401 cagcctcttc gtaacttgac cattctggat ctaagcaaca acaacatagc caacataaat
2461 gatgacatgt tggagggtct tgagaaacta gaaattctcg atttgcagca taacaactta
2521 gcacggctct ggaaacacgc aaaccctggt ggtcccattt atttcctaaa gggtctgtct
2581 cacctccaca tccttaactt ggagtccaac ggctttgacg agatcccagt tgaggtcttc
2641 aaggatttat ttgaactaaa gatcatcgat ttaggattga ataatttaaa cacacttcca
2701 gcatctgtct ttaataatca ggtgtctcta aagtcattga accttcagaa gaatctcata
2761 acatccgttg agaagaaggt tttcgggcca gctttcagga acctgactga gttagatatg
2821 cgctttaatc cctttgattg cacgtgtgaa agtattgcct ggtttgttaa ttggattaac
2881 gagacccata ccaacatccc tgagctgtca agccactacc tttgcaacac tccacctcac
2941 tatcatgggt tcccagtgag actttttgat acatcatctt gcaaagacag tgcccccttt
3001 gaactctttt tcatgatcaa taccagtatc ctgttgattt ttatctttat tgtacttctc
3061 atccactttg agggctggag gatatctttt tattggaatg tttcagtaca tcgagttctt
3121 ggtttcaaag aaatagacag acagacagaa cagtttgaat atgcagcata tataattcat
3181 gcctataaag ataaggattg ggtctgggaa catttctctt caatggaaaa ggaagaccaa
3241 tctctcaaat tttgtctgga agaaagggac tttgaggcgg gtgtttttga actagaagca
3301 attgttaaca gcatcaaaag aagcagaaaa attatttttg ttataacaca ccatctatta
3361 aaagacccat tatgcaaaag attcaaggta catcatgcag ttcaacaagc tattgaacaa
3421 aatctggatt ccattatatt ggttttcctt gaggagattc cagattataa actgaaccat
3481 gcactctgtt tgcgaagagg aatgtttaaa tctcactgca tcttgaactg gccagttcag
3541 aaagaacgga taggtgcctt tcgtcataaa ttgcaagtag cacttggatc caaaaactct
3601 gtacatctcg agatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
3661 gtcgagctgg acggcgacgt aaacggccac aggttcagcg tgtccggcga gggcgagggc
3721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
3781 ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcag ccgctacccc
3841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
3901 cgtaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
3961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
4021 atcctggggc acaagctgga gtacaactac atcagccaca acgtctatat caccgccgac
4081 aagcagaaga acggcatcaa ggcccacttc aagatccgcc acaacatcga ggacggcagc
4141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
4201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
4261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
4321 ctgtacaagt aatctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg
4381 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
4441 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
4501 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
4561 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
4621 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
4681 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
4741 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
4801 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
4861 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
4921 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
4981 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
5041 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
5101 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc caggcaggca gaagtatgca
5161 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
5221 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
5281 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
5341 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
5401 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
5461 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
5521 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
5581 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
5641 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
5701 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
5761 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
5821 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
5881 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
5941 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
6001 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
6061 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
6121 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
6181 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
6241 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
6301 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
6361 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
6421 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
6481 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
6541 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
6601 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
6661 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
6721 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
6781 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
6841 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
6901 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
6961 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
7021 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
7081 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
7141 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
7201 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
7261 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
7321 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
7381 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
7441 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
7501 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
7561 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
7621 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
7681 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
7741 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
7801 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
7861 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
7921 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
7981 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
8041 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
8101 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
8161 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
8221 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
8281 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
8341 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
8401 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
8461 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
8521 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
8581 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
8641 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
8701 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
8761 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
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