| 质粒名称: | pNL-EGFP/CMV/WPREdU3 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 17579 |
| 存储实验室: | Jakob Reiser |
|
载体骨架基本信息 |
|
| 载体名称: | pUC18 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 8998 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 慢病毒载体 |
| 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: | EGFP |
| 物种来源: | Aequorea victoria |
| 插入大小: | 761 bp |
| 基因突变: | -- |
| GenBank编号 | U55763 |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NheI、AfeI、AgeI（销毁后未知） |
| 3' 克隆位点: | XhoI，BsrGI（销毁时未知） |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pNL-EGFP/CMV/WPREdU3 was a gift from Jakob Reiser (Addgene plasmid # 17579 ; http://n2t.net/addgene:17579 ; RRID:Addgene_17579) |
| 参考文献部分: | Optimized lentiviral transduction of mouse bone marrow-derived mesenchymal stem cells. Ricks DM, Kutner R, Zhang XY, Welsh DA, Reiser J. Stem Cells Dev. 2008 Jun . 17(3):441-50. 10.1089/scd.2007.0194 PubMed 18513160 |
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 pNL-EGFP/CMV/WPREdU3 9759 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pNL-EGFP/CMV/WPREdU3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9759)
AUTHORS Ricks DM, Kutner R, Zhang XY, Welsh DA, Reiser J
TITLE Optimized lentiviral transduction of mouse bone marrow-derived
mesenchymal stem cells.
JOURNAL Stem Cells Dev. 2008 Jun . 17(3):441-50.
PUBMED 18513160
REFERENCE 2 (bases 1 to 9759)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9759
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 377..634
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin 1261..1396
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 1323..1342
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
misc_feature 1674..1907
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
enhancer 2406..2709
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2710..2913
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2864..2884
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 2952..2961
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2958..3674
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3003..3024)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3264..3283)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3611..3632
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 3694..4282
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3747..3767)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4165..4176)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 4409..4589
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(6231..6248)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6402..6990)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6482..6501)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7161..8021)
/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 7784..7803
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8022..8126)
/gene="bla"
/label=AmpR promoter
primer_bind 8194..8212
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(8263..8285)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 8371..8509
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 8424..8443
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 tggaagggct aatttggtcc caaaaaagac aagagatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc agatatccac
121 tgacctttgg atggtgcttc aagttagtac cagttgaacc agagcaagta gaagaggcca
181 aataaggaga gaagaacagc ttgttacacc ctatgagcca gcatgggatg gaggacccgg
241 agggagaagt attagtgtgg aagtttgaca gcctcctagc atttcgtcac atggcccgag
301 agctgcatcc ggagtactac aaagactgct gacatcgagc tttctacaag ggactttccg
361 ctggggactt tccagggagg tgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 gctacatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgctca aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagtaaa gccagaggag atctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcggta ttaagcgggg gagaattaga taaatgggaa
841 aaaattcggt taaggccagg gggaaagaaa caatataaac taaaacatat agtatgggca
901 agcagggagc tagaacgatt cgcagttaat cctggccttt tagagacatc agaaggctgt
961 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
1021 ttatataata caatagcagt cctctattgt gtgcatcaaa ggatagatgt aaaagacacc
1081 aaggaagcct tagataagat agaggaagag caaaacaaaa gtaagaaaaa ggcacagcaa
1141 gcagcagctg acacaggaaa caacagccag gtcagccaaa attaccctat agtgcagaac
1201 ctccaggggc aaatggtaca tcaggccata tcacctagaa ctttaaatat gactccgccc
1261 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
1321 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
1381 ggcttttttg gaggcctagg cttttgcaaa tgcataaaag aaaagggggg attggggggt
1441 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
1501 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccac
1561 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
1621 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
1681 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
1741 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
1801 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
1861 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
1921 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
1981 tctctggaac agatttggaa taacatgacc tggatggagt gggacagaga aattaacaat
2041 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
2101 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
2161 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
2221 gtttttgctg tactttctgt agtgaataga gttaggcagg gatattcacc attatcgttt
2281 cagacccacc tcccaatccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
2341 ggagagagag acagagacag atccattcga ttagtgaacg gatctagtta actagatcta
2401 gagtccgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
2461 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
2521 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
2581 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
2641 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
2701 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
2761 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
2821 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
2881 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
2941 cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
3001 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
3061 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
3121 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
3181 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
3241 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
3301 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
3361 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
3421 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
3481 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
3541 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
3601 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
3661 acgagctgta caagtccgga ctcagatctc gagaatcaac ctctggatta caaaatttgt
3721 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
3781 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
3841 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
3901 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
3961 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
4021 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
4081 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
4141 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
4201 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
4261 tccctttggg ccgcctcccc gcctgggtac ctttaagacc aatgacttac aaggcagctg
4321 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaaa
4381 gaagacaaag ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg
4441 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
4501 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
4561 cttttagtca gtgtggaaaa tctctagcac cccccaggag gtagaggttg cagtgagcca
4621 agatcgcgcc actgcattcc agcctgggca agaaaacaag actgtctaaa ataataataa
4681 taagttaagg gtattaaata tatttataca tggaggtcat aaaaatatat atatttgggc
4741 tgggcgcagt ggctcacacc tgcgcccggc cctttgggag gccgaggcag gtggatcacc
4801 tgagtttggg agttccagac cagcctgacc aacatggaga aaccccttct ctgtgtattt
4861 ttagtagatt ttattttatg tgtattttat tcacaggtat ttctggaaaa ctgaaactgt
4921 ttttcctcta ctctgatacc acaagaatca tcagcacaga ggaagacttc tgtgatcaaa
4981 tgtggtggga gagggaggtt ttcaccagca catgagcagt cagttctgcc gcagactcgg
5041 cgggtgtcct tcggttcagt tccaacaccg cctgcctgga gagaggtcag accacagggt
5101 gagggctcag tccccaagac ataaacaccc aagacataaa cacccaacag gtccaccccg
5161 cctgctgccc aggcagagcc gattcaccaa gacgggaatt aggatagaga aagagtaagt
5221 cacacagagc cggctgtgcg ggagaacgga gttctattat gactcaaatc agtctcccca
5281 agcattcggg gatcagagtt tttaaggata acttagtgtg tagggggcca gtgagttgga
5341 gatgaaagcg tagggagtcg aaggtgtcct tttgcgccga gtcagttcct gggtgggggc
5401 cacaagatcg gatgagccag tttatcaatc cgggggtgcc agctgatcca tggagtgcag
5461 ggtctgcaaa atatctcaag cactgattga tcttaggttt tacaatagtg atgttacccc
5521 aggaacaatt tggggaaggt cagaatcttg tagcctgtag ctgcatgact cctaaaccat
5581 aatttctttt ttgttttttt ttttttattt ttgagacagg gtctcactct gtcacctagg
5641 ctggagtgca gtggtgcaat cacagctcac tgcagcctca acgtcgtaag ctcaagcgat
5701 cctcccacct cagcctgcct ggtagctgag actacaagcg acgccccagt taatttttgt
5761 atttttggta gaggcagcgt tttgccgtgt ggccctggct ggtctcgaac tcctgggctc
5821 aagtgatcca gcctcagcct cccaaagtgc tgggacaacc ggggccagtc actgcacctg
5881 gccctaaacc ataatttcta atcttttggc taatttgtta gtcctacaaa ggcagtctag
5941 tccccaggca aaaagggggt ttgtttcggg aaagggctgt tactgtcttt gtttcaaact
6001 ataaactaag ttcctcctaa acttagttcg gcctacaccc aggaatgaac aaggagagct
6061 tggaggttag aagcacgatg gaattggtta ggtcagatct ctttcactgt ctgagttata
6121 attttgcaat ggtggttcaa agactgcccg cttctgacac cagtcgctgc attaatgaat
6181 cggccaacgc gcggggagag gcggtttgcg tattggcgct cttccgcttc ctcgctcact
6241 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
6301 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
6361 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
6421 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
6481 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
6541 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
6601 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
6661 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
6721 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
6781 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
6841 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
6901 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
6961 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
7021 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
7081 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
7141 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
7201 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
7261 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
7321 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
7381 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
7441 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
7501 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
7561 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
7621 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
7681 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
7741 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
7801 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
7861 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
7921 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
7981 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
8041 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
8101 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
8161 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
8221 cttcaagaac tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
8281 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
8341 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
8401 cggagtgtac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
8461 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
8521 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
8581 ggggaggcag agattgcagt aagctgagat cgcagcactg cactccagcc tgggcgacag
8641 agtaagactc tgtctcaaaa ataaaataaa taaatcaatc agatattcca atcttttcct
8701 ttatttattt atttattttc tattttggaa acacagtcct tccttattcc agaattacac
8761 atatattcta tttttcttta tatgctccag ttttttttag accttcacct gaaatgtgtg
8821 tatacaaaat ctaggccagt ccagcagagc ctaaaggtaa aaaataaaat aataaaaaat
8881 aaataaaatc tagctcactc cttcacatca aaatggagat acagctgtta gcattaaata
8941 ccaaataacc catcttgtcc tcaataattt taagcgcctc tctccaccac atctaactcc
9001 tgtcaaaggc atgtgcccct tccgggcgct ctgctgtgct gccaaccaac tggcatgtgg
9061 actctgcagg gtccctaact gccaagcccc acagtgtgcc ctgaggctgc cccttccttc
9121 tagcggctgc ccccactcgg ctttgctttc cctagtttca gttacttgcg ttcagccaag
9181 gtctgaaact aggtgcgcac agagcggtaa gactgcgaga gaaagagacc agctttacag
9241 ggggtttatc acagtgcacc ctgacagtcg tcagcctcac agggggttta tcacattgca
9301 ccctgacagt cgtcagcctc acagggggtt tatcacagtg cacccttaca atcattccat
9361 ttgattcaca atttttttag tctctactgt gcctaacttg taagttaaat ttgatcagag
9421 gtgtgttccc agaggggaaa acagtatata cagggttcag tactatcgca tttcaggcct
9481 ccacctgggt cttggaatgt gtcccccgag gggtgatgac tacctcagtt ggatctccac
9541 aggtcacagt gacacaagat aaccaagaca cctcccaagg ctaccacaat gggccgccct
9601 ccacgtgcac atggccggag gaactgccat gtcggaggtg caagcacacc tgcgcatcag
9661 agtccttggt gtggagggag ggaccagcgc agcttccagc catccacctg atgaacagaa
9721 cctagggaaa gccccagttc tacttacacc aggaaaggc
//
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