| 质粒名称: | pCEP4 Pig4 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 16498 |
| 存储实验室: | Bert Vogelstein |
|
载体骨架基本信息 |
|
| 载体名称: | pCEP4 |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄西林 |
| 空载体大小: | 10400bp |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | 潮霉素 |
| 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: | Pig4 |
| 备用名称: | SAA1、serum amyloid A-1 protein preproprotein |
| 物种来源: | 智人（人） |
| 插入大小: | 450bp |
| Entrez基因: | SAA1 (a.k.a. PIG4, SAA, SAA2, TP53I4 |
| 启动子 | CMV |
| 标签/融合蛋白: | HA标签变体-Yaydvpdya（插入件上的C端子） |
| 克隆方法: | 限制性酶 |
| 5' 克隆位点: | 八木（未销毁） |
| 3' 克隆位点: | XhoI (not destroyed) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | EBV-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pCEP4 Pig4 was a gift from Bert Vogelstein (Addgene plasmid # 16498 ; http://n2t.net/addgene:16498 ; RRID:Addgene_16498) |
| 参考文献部分: |
A model for p53-induced apoptosis. Polyak K, Xia Y, Zweier JL, Kinzler KW, Vogelstein B. Nature. 1997 Sep 18. 389(6648):300-5. 10.1038/38525 PubMed 9305847 |
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载体基本信息
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载体简介
载体序列
LOCUS pCEP4 Pig4 10024 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCEP4 Pig4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10024) AUTHORS Polyak K, Xia Y, Zweier JL, Kinzler KW, Vogelstein B TITLE A model for p53-induced apoptosis. JOURNAL Nature. 1997 Sep 18. 389(6648):300-5. PUBMED 9305847 REFERENCE 2 (bases 1 to 10024) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10024 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 110..132 /label=pGEX 3' /note="pGEX vectors, reverse primer" polyA_signal 275..409 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(312..331) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 366..385 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(839..858) /label=T7 /note="T7 promoter, forward primer" promoter complement(840..858) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(915..939) /label=LNCX /note="Human CMV promoter, forward primer" promoter complement(936..1139) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(965..985) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 1140..1519 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" polyA_signal 2173..2221 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" primer_bind 2246..2265 /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" CDS complement(2263..3300) /codon_start=1 /product="hygromycin B phosphotransferase" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE MGEAN" promoter complement(3343..3488) /label=HSV TK promoter /note="herpes simplex virus thymidine kinase promoter" rep_origin complement(3919..4507) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3999..4018) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4678..5538) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5301..5320 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5539..5643) /gene="bla" /label=AmpR promoter primer_bind 5711..5729 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" rep_origin 7785..9574 /label=oriP /note="Epstein-Barr virus oriP replication origin (Yates et al., 2000)" ORIGIN 1 tctagagtcg acctcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga 61 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc 121 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagccgga tcataatcag 181 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa 241 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg 301 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 361 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc ttagctagcg 421 taatctggaa catcgtaagc gtaatctgga acatcgtacc ccccgtattt ctcagacagg 481 ccagcaggtc ggaagtgatt ggggtctttg ccactcctgc cccattcatt ggcagcctga 541 tcagccagcg agtcctccgc accatggcca aagaatctct ggatattctc tctggcatcg 601 ctgatcactt ctgcagccca ggcaccccca ggtccccttt tggcagcatc atagttcccc 661 cgagcatgga agtatttgtc tgagccgatg taattggctt ctctcatgtc agagtaggct 721 ctccacatgt cccgagcccc atcaaaagcc tcgccaagga acgaaaagaa gcttcggctg 781 ctgacaccca ggaccaagga gcagaaaacc aggcccgtga gaagctccat ggtggtctcc 841 ctatagtgag tcgtattagg atccctcgag cggccgctag caagcttgct agcagctggt 901 acccagcttc tagagatctg acggttcact aaacgagctc tgcttatata gacctcccac 961 cgtacacgcc taccgcccat ttgcgtcaac ggggcggggt tattacgaca ttttggaaag 1021 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga 1081 aatccccgtg agtcaaaccg ctatccacgc ccattggtgt actgccaaaa ccgcatcacc 1141 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccgtaaggtc 1201 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgga 1261 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc 1321 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac 1381 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta 1441 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa 1501 ctagtcaata atcaatgtca acatggcggt catattggac atgagccaat ataaatgtac 1561 atattatgat atagatacaa cgtatgcaat ggccaatagc caatattgat ttatgctata 1621 taaccaatga ctaatatggc taattgccaa tattgattca atgtatagat cttccatacc 1681 taccagttct gcgcctgcag caatggcaac aacgttgccc ggatctgcga tgataagctg 1741 tcaaacatga gaattggtcg actagcttgg cacgccagaa atccgcgcgg tggtttttgg 1801 gggtcggggg tgtttggcag ccacagacgc ccggtgttcg tgtcgcgcca gtacatgcgg 1861 tccatgccca ggccatccaa aaaccatggg tctgtctgct cagtccagtc gtggacctga 1921 ccccacgcaa cgcccaaaat aataaccccc acgaaccata aaccattccc catgggggac 1981 cccgtcccta acccacgggg ccagtggcta tggcagggcc tgccgccccg acgttggctg 2041 cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg 2101 cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa 2161 ccccgcgttt atgaacaaac gacccaacac ccgtgcgttt tattctgtct ttttattgcc 2221 gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag cctcccccat 2281 ctcccgatcc ggacgagtgc tggggcgtcg gtttccacta tcggcgagta cttctacaca 2341 gccatcggtc cagacggccg cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc 2401 tccggatcgg acgattgcgt cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc 2461 gtcaaccaag ctctgataga gttggtcaag accaatgcgg agcatatacg cccggagccg 2521 cggcgatcct gcaagctccg gatgcctccg ctcgaagtag cgcgtctgct gctccataca 2581 agccaaccac ggcctccaga agaagatgtt ggcgacctcg tattgggaat ccccgaacat 2641 cgcctcgctc cagtcaatga ccgctgttat gcggccattg tccgtcagga cattgttgga 2701 gccgaaatcc gcgtgcacga ggtgccggac ttcggggcag tcctcggccc aaagcatcag 2761 ctcatcgaga gcctgcgcga cggacgcact gacggtgtcg tccatcacag tttgccagtg 2821 atacacatgg ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt attgaccgat 2881 tccttgcggt ccgaatgggc cgaacccgct cgtctggcta agatcggccg cagcgatcgc 2941 atccatggcc tccgcgaccg gctgcagaac agcgggcagt tcggtttcag gcaggtcttg 3001 caacgtgaca ccctgtgcac ggcgggagat gcaataggtc aggctctcgc tgaattcccc 3061 aatgtcaagc acttccggaa tcgggagcgc ggccgatgca aagtgccgat aaacataacg 3121 atctttgtag aaaccatcgg cgcagctatt tacccgcagg acatatccac gccctcctac 3181 atcgaagctg aaagcacgag attcttcgcc ctccgagagc tgcatcaggt cggagacgct 3241 gtcgaacttt tcgatcagaa acttctcgac agacgtcgcg gtgagttcag gctttttcat 3301 atctcattgc cccccgggat ctgcggcacg ctgttgacgc tgttaagcgg gtcgctgcag 3361 ggtcgctcgg tgttcgaggc cacacgcgtc accttaatat gcgaagtgga cctcggaccg 3421 cgccgccccg actgcatctg cgtgttcgaa ttcgccaatg acaagacgct gggcggggtt 3481 tgtgtcatca tagaactaaa gacatgcaaa tatatttctt ccggggacac cgccagcaaa 3541 cgcgagcaac gggccacggg gatgaagcag ggcggcacct cgctaacgga ttcaccactc 3601 caagaattgg agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc 3661 agaacatatc catcgcgtcc gccatctcca gcagccgcac gcggcgcatc tcggggccga 3721 cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct tccccattat 3781 gattcttctc gcttccggcg gcatcgggat gcccgcgttg caggccatgc tgtccaggca 3841 ggtagatgac gaccatcagg gacagcttca aggccagcaa aaggccagga accgtaaaaa 3901 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 3961 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 4021 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 4081 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 4141 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 4201 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 4261 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 4321 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 4381 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 4441 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 4501 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 4561 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 4621 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 4681 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 4741 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 4801 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 4861 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 4921 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 4981 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 5041 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 5101 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 5161 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 5221 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 5281 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat 5341 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 5401 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 5461 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 5521 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 5581 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 5641 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt 5701 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattct catgtttgac 5761 agcttatcat cgataagctg atcctcacag gccgcaccca gcttttcttc cgttgcccca 5821 gtagcatctc tgtctggtga ccttgaagag gaagaggagg ggtcccgaga atccccatcc 5881 ctaccgtcca gcaaaaaggg ggacgaggaa tttgaggcct ggcttgaggc tcaggacgca 5941 aatcttgagg atgttcagcg ggagttttcc gggctgcgag taattggtga tgaggacgag 6001 gatggttcgg aggatgggga attttcagac ctggatctgt ctgacagcga ccatgaaggg 6061 gatgagggtg ggggggctgt tggagggggc aggagtctgc actccctgta ttcactgagc 6121 gtcgtctaat aaagatgtct attgatctct tttagtgtga atcatgtctg acgaggggcc 6181 aggtacagga cctggaaatg gcctaggaga gaagggagac acatctggac cagaaggctc 6241 cggcggcagt ggacctcaaa gaagaggggg tgataaccat ggacgaggac ggggaagagg 6301 acgaggacga ggaggcggaa gaccaggagc cccgggcggc tcaggatcag ggccaagaca 6361 tagagatggt gtccggagac cccaaaaacg tccaagttgc attggctgca aagggaccca 6421 cggtggaaca ggagcaggag caggagcggg aggggcagga gcaggagggg caggagcagg 6481 aggaggggca ggagcaggag gaggggcagg agcaggaggg gcaggagcag gaggtggagg 6541 ccggggtcga ggaggcagtg gaggccgggg tcgaggaggt agtggaggcc ggggtcgagg 6601 aggtagtgga ggccgccggg gtagaggacg tgaaagagcc agggggggaa gtcgtgaaag 6661 agccaggggg agaggtcgtg gacgtggaga aaagaggccc aggagtccca gtagtcagtc 6721 atcatcatcc gggtctccac cgcgcaggcc ccctccaggt agaaggccat ttttccaccc 6781 tgtaggggaa gccgattatt ttgaatacca ccaagaaggt ggcccagatg gtgagcctga 6841 cgtgcccccg ggagcgatag agcagggccc cgcagatgac ccaggagaag gcccaagcac 6901 tggaccccgg ggtcagggtg atggaggcag gcgcaaaaaa ggagggtggt ttggaaagca 6961 tcgtggtcaa ggaggttcca acccgaaatt tgagaacatt gcagaaggtt taagagctct 7021 cctggctagg agtcacgtag aaaggactac cgacgaagga acttgggtcg ccggtgtgtt 7081 cgtatatgga ggtagtaaga cctcccttta caacctaagg cgaggaactg cccttgctat 7141 tccacaatgt cgtcttacac cattgagtcg tctccccttt ggaatggccc ctggacccgg 7201 cccacaacct ggcccgctaa gggagtccat tgtctgttat ttcatggtct ttttacaaac 7261 tcatatattt gctgaggttt tgaaggatgc gattaaggac cttgttatga caaagcccgc 7321 tcctacctgc aatatcaggg tgactgtgtg cagctttgac gatggagtag atttgcctcc 7381 ctggtttcca cctatggtgg aaggggctgc cgcggagggt gatgacggag atgacggaga 7441 tgaaggaggt gatggagatg agggtgagga agggcaggag tgatgtaact tgttaggaga 7501 cgccctcaat cgtattaaaa gccgtgtatt cccccgcact aaagaataaa tccccagtag 7561 acatcatgcg tgctgttggt gtatttctgg ccatctgtct tgtcaccatt ttcgtcctcc 7621 caacatgggg caattgggca tacccatgtt gtcacgtcac tcagctccgc gctcaacacc 7681 ttctcgcgtt ggaaaacatt agcgacattt acctggtgag caatcagaca tgcgacggct 7741 ttagcctggc ctccttaaat tcacctaaga atgggagcaa ccagcaggaa aaggacaagc 7801 agcgaaaatt cacgccccct tgggaggtgg cggcatatgc aaaggatagc actcccactc 7861 tactactggg tatcatatgc tgactgtata tgcatgagga tagcatatgc tacccggata 7921 cagattagga tagcatatac tacccagata tagattagga tagcatatgc tacccagata 7981 tagattagga tagcctatgc tacccagata taaattagga tagcatatac tacccagata 8041 tagattagga tagcatatgc tacccagata tagattagga tagcctatgc tacccagata 8101 tagattagga tagcatatgc tacccagata tagattagga tagcatatgc tatccagata 8161 tttgggtagt atatgctacc cagatataaa ttaggatagc atatactacc ctaatctcta 8221 ttaggatagc atatgctacc cggatacaga ttaggatagc atatactacc cagatataga 8281 ttaggatagc atatgctacc cagatataga ttaggatagc ctatgctacc cagatataaa 8341 ttaggatagc atatactacc cagatataga ttaggatagc atatgctacc cagatataga 8401 ttaggatagc ctatgctacc cagatataga ttaggatagc atatgctatc cagatatttg 8461 ggtagtatat gctacccatg gcaacattag cccaccgtgc tctcagcgac ctcgtgaata 8521 tgaggaccaa caaccctgtg cttggcgctc aggcgcaagt gtgtgtaatt tgtcctccag 8581 atcgcagcaa tcgcgcccct atcttggccc gcccacctac ttatgcaggt attccccggg 8641 gtgccattag tggttttgtg ggcaagtggt ttgaccgcag tggttagcgg ggttacaatc 8701 agccaagtta ttacaccctt attttacagt ccaaaaccgc agggcggcgt gtgggggctg 8761 acgcgtgccc ccactccaca atttcaaaaa aaagagtggc cacttgtctt tgtttatggg 8821 ccccattggc gtggagcccc gtttaatttt cgggggtgtt agagacaacc agtggagtcc 8881 gctgctgtcg gcgtccactc tctttcccct tgttacaaat agagtgtaac aacatggttc 8941 acctgtcttg gtccctgcct gggacacatc ttaataaccc cagtatcata ttgcactagg 9001 attatgtgtt gcccatagcc ataaattcgt gtgagatgga catccagtct ttacggcttg 9061 tccccacccc atggatttct attgttaaag atattcagaa tgtttcattc ctacactagt 9121 atttattgcc caaggggttt gtgagggtta tattggtgtc atagcacaat gccaccactg 9181 aaccccccgt ccaaatttta ttctgggggc gtcacctgaa accttgtttt cgagcacctc 9241 acatacacct tactgttcac aactcagcag ttattctatt agctaaacga aggagaatga 9301 agaagcaggc gaagattcag gagagttcac tgcccgctcc ttgatcttca gccactgccc 9361 ttgtgactaa aatggttcac taccctcgtg gaatcctgac cccatgtaaa taaaaccgtg 9421 acagctcatg gggtgggaga tatcgctgtt ccttaggacc cttttactaa ccctaattcg 9481 atagcatatg cttcccgttg ggtaacatat gctattgaat tagggttagt ctggatagta 9541 tatactacta cccgggaagc atatgctacc cgtttagggt taacaagggg gccttataaa 9601 cactattgct aatgccctct tgagggtccg cttatcggta gctacacagg cccctctgat 9661 tgacgttggt gtagcctccc gtagtcttcc tgggcccctg ggaggtacat gtcccccagc 9721 attggtgtaa gagcttcagc caagagttac acataaaggc aatgttgtgt tgcagtccac 9781 agactgcaaa gtctgctcca ggatgaaagc cactcagtgt tggcaaatgt gcacatccat 9841 ttataaggat gtcaactaca gtcagagaac ccctttgtgt ttggtccccc cccgtgtcac 9901 atgtggaaca gggcccagtt ggcaagttgt accaaccaac tgaagggatt acatgcactg 9961 ccccgcgaag aaggggcaga gatgtcgtag tcaggtttag ttcgtccggg gcggggatcg 10021 atcc //
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