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AAVS1-Neo-M2rtTA载体质粒基本信息
AAVS1-Neo-M2rtTA质粒图谱载体图谱和AAVS1-Neo-M2rtTA载体序列质粒序列多克隆位点信息
AAVS1-Neo-M2rtTA质粒载体简介
AAVS1-Neo-M2rtTA质粒序列载体序列
LOCUS AAVS1-Neo-M2rtTA 10080 bp DNA circular SYN 12-JUL-2023 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10080) AUTHORS Biofeng TITLE Direct Submission JOURNAL Exported Jul 12, 2023 from SnapGene 6.0.2 https://www.biofeng.com FEATURES Location/Qualifiers source 1..10080 /mol_type="other DNA" /organism="synthetic DNA construct" misc_feature 33..836 /label=HA-L /note="left homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene" misc_feature 843..868 /label=SA /note="splice acceptor site" CDS 892..945 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 961..1764 /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" enhancer 2389..2692 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2694..2971 /label=chicken beta-actin promoter intron 2973..3990 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" CDS 4090..4836 /codon_start=1 /product="improved tetracycline-controlled transactivator" /label=rtTA-Advanced /note="In the Tet-On(R) system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline." /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA DALDDFDLDMLPADALDDFDLDMLPG" polyA_signal 4846..4980 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" misc_feature 5310..6146 /label=HA-R /note="right homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene" promoter complement(6187..6205) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6212..6228) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 6366..6668 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" CDS complement(8065..8925) /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" rep_origin 9049..9637 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 9925..9946 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 9961..9991 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9999..10015 /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)." primer_bind 10023..10039 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 10060..10078 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 ctagtcctgc aggtttaaac gaattcgccc tttgctttct ctgaccagca ttctctcccc 61 tgggcctgtg ccgctttctg tctgcagctt gtggcctggg tcacctctac ggctggccca 121 gatccttccc tgccgcctcc ttcaggttcc gtcttcctcc actccctctt ccccttgctc 181 tctgctgtgt tgctgcccaa ggatgctctt tccggagcac ttccttctcg gcgctgcacc 241 acgtgatgtc ctctgagcgg atcctccccg tgtctgggtc ctctccgggc atctctcctc 301 cctcacccaa ccccatgccg tcttcactcg ctgggttccc ttttccttct ccttctgggg 361 cctgtgccat ctctcgtttc ttaggatggc cttctccgac ggatgtctcc cttgcgtccc 421 gcctcccctt cttgtaggcc tgcatcatca ccgtttttct ggacaacccc aaagtacccc 481 gtctccctgg ctttagccac ctctccatcc tcttgctttc tttgcctgga caccccgttc 541 tcctgtggat tcgggtcacc tctcactcct ttcatttggg cagctcccct acccccctta 601 cctctctagt ctgtgctagc tcttccagcc ccctgtcatg gcatcttcca ggggtccgag 661 agctcagcta gtcttcttcc tccaacccgg gcccctatgt ccacttcagg acagcatgtt 721 tgctgcctcc agggatcctg tgtccccgag ctgggaccac cttatattcc cagggccggt 781 taatgtggct ctggttctgg gtacttttat ctgtcccctc caccccacag tggggcaagc 841 ttctgacctc ttctcttcct cccacagggc ctcgagagat ctggcagcgg agagggcaga 901 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctaggct cgagacgcgt 961 atgggatcgg ccattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 1021 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 1081 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 1141 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 1201 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 1261 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 1321 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 1381 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 1441 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc 1501 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 1561 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 1621 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 1681 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 1741 cgccttcttg acgagttctt ctgaggggat ccgctgtaag tctgcagaaa ttgatgatct 1801 attaaacaat aaagatgtcc actaaaagga agtttttctg cagaaattga tgatctatta 1861 aacaataaag atgtccacta aaatgaagtt ttttactgtc atactttgtt aagaagggtg 1921 agaacagagt acctacattt tgaatggaag gattggagct acgggggtgg gggtggggtg 1981 ggattagata aatgcctgct ctttactgaa ggctctttac tattgcttta tgataatgtt 2041 tcatagttgg atatcataat ttaaacaagc aaaaccaaat taagggccag ctcattcctc 2101 ccactcatga tctatagatc tatagatctc tcgtgggatc attgtttttc tcttgattcc 2161 cactttgtgg ttctaagtac tgtggtttcc aaatgtgtca gtttcatagc ctgaagaacg 2221 agatcagcag cctctgttcc acatacactt cattctcagt attgttttgc caagttctaa 2281 ttccatcaga agctgggagc tccaccgcgg tggcggccgc tctagaacta gttattaata 2341 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact 2401 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat 2461 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta 2521 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc 2581 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg 2641 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt 2701 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta 2761 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc 2821 gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc 2881 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg 2941 gccctataaa aagcgaagcg cgcggcgggc ggggagtcgc tgcgacgctg ccttcgcccc 3001 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc 3061 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa 3121 tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct 3181 ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg 3241 tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg 3301 ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc 3361 tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg 3421 ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc 3481 ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg 3541 gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg 3601 ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc 3661 cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt 3721 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg 3781 tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt 3841 ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga 3901 cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac 3961 catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg 4021 tctcatcatt ttggcaaaga attcggcttg atcgaagcta gtggatcccc cgggctgcag 4081 gaattcacca tgtctagact ggacaagagc aaagtcataa acggcgctct ggaattactc 4141 aatggagtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag 4201 cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc 4261 gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc atggcaagac 4321 tttctgcgga acaacgccaa gtcattccgc tgtgctctcc tctcacatcg cgacggggct 4381 aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc 4441 gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg 4501 ggccacttta cactgggctg cgtattggag gaacaggagc atcaagtagc aaaagaggaa 4561 agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat tgagctgttc 4621 gaccggcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg 4681 gagaaacagc taaagtgcga aagcggcggg ccggccgacg cccttgacga ttttgactta 4741 gacatgctcc cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct 4801 cttgacgatt ttgaccttga catgctcccc gggtaactaa gtaaggatcc agacatgata 4861 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt 4921 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt 4981 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt 5041 taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcctgc aagcctcgtc 5101 gtctggccgg accacgctat ctgtgcaagg tccccggacg cgcgctccat gagcagagcg 5161 cccgccgccg aggcaagact cgggcggcgc cctgcccgtc ccaccaggtc aacaggcggt 5221 aaccggcctc ttcatcggga atgcgcgcga ccttcagcat cgccggcatg tcccctggcg 5281 gacgggaagt atcagctcga ccaagcttta ctagggacag gattggtgac agaaaagccc 5341 catccttagg cctcctcctt cctagtctcc tgatattggg tctaaccccc acctcctgtt 5401 aggcagattc cttatctggt gacacacccc catttcctgg agccatctct ctccttgcca 5461 gaacctctaa ggtttgctta cgatggagcc agagaggatc ctgggaggga gagcttggca 5521 gggggtggga gggaaggggg ggatgcgtga cctgcccggt tctcagtggc caccctgcgc 5581 taccctctcc cagaacctga gctgctctga cgcggctgtc tggtgcgttt cactgatcct 5641 ggtgctgcag cttccttaca cttcccaaga ggagaagcag tttggaaaaa caaaatcaga 5701 ataagttggt cctgagttct aactttggct cttcaccttt ctagtcccca atttatattg 5761 ttcctccgtg cgtcagtttt acctgtgaga taaggccagt agccagcccc gtcctggcag 5821 ggctgtggtg aggagggggg tgtccgtgtg gaaaactccc tttgtgagaa tggtgcgtcc 5881 taggtgttca ccaggtcgtg gccgcctcta ctccctttct ctttctccat ccttctttcc 5941 ttaaagagtc cccagtgcta tctgggacat attcctccgc ccagagcagg gtcccgcttc 6001 cctaaggccc tgctctgggc ttctgggttt gagtccttgg caagcccagg agaggcgctc 6061 aggcttccct gtcccccttc ctcgtccacc atctcatgcc cctggctctc ctgccccttc 6121 cctacagggg ttcctggctc tgctctaagg gcaagggcga attcgcggcc gctaaattca 6181 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact 6241 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct 6301 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctatacg 6361 tacggcagtt taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg 6421 tacagagtga tattattgac acgccggggc gacggatggt gatccccctg gccagtgcac 6481 gtctgctgtc agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa 6541 gctggcgcat gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag 6601 tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg 6661 gaatataaat gtcaggcatg agattatcaa aaaggatctt cacctagatc cttttcacgt 6721 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct 6781 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc 6841 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc 6901 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga 6961 tctgatggcg caggggatca agctctgatc aagagacagg atgaggatcg tttcgcatga 7021 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 7081 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 7141 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag 7201 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 7261 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 7321 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 7381 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 7441 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 7501 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg 7561 aggatctcgt cgtgacccat gcatggcgat gcctgcttgc cgaatatcat ggtggaaaat 7621 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac 7681 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc 7741 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt 7801 gacgagttct tctgaattat taacgcttac aatttcctga tgcggtattt tctccttacg 7861 catctgtgcg gtatttcaca ccgcatcagg tggcactttt cggggaaatg tgcgcggaac 7921 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gattatcaaa 7981 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 8041 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 8101 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 8161 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 8221 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 8281 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 8341 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 8401 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 8461 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 8521 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 8581 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 8641 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 8701 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 8761 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 8821 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 8881 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 8941 atattattga agcatttatc agggttattg tctcatgacc aaaatccctt aacgtgagtt 9001 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 9061 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 9121 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 9181 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 9241 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 9301 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 9361 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 9421 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 9481 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 9541 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 9601 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 9661 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 9721 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac 9781 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc 9841 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa 9901 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc 9961 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca 10021 cacaggaaac agctatgacc atgattacgc caagctcaga attaaccctc actaaaggga //
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AAVS1-Neo-M2rtTA质粒载体应用举例
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