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pH-ABE8e-spG

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聯系方式:I47-825O-882O

相關技術服務:質粒構建    基因合成    質粒大提

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pH-ABE8e-spG載體質粒基本信息

質粒名稱: pH-ABE8e-spG
出品公司: Addgene
目錄編號: 200775
存儲實驗室: Caixia Gao

載體骨架基本信息
載體名稱: pHUE411 (modified from pCambia)
載體出品公司: Clontech
完整質粒大小: 17211bp
載體類型: Plant Expression, CRISPR
篩選標記: Hygromycin
載體抗性: Kanamycin, 50 μg/mL
生長溫度: 37 ℃
宿主菌: DH5a
拷貝數: 高拷貝

插入基因信息
插入基因名稱: nCas9-spG(D10A)-TadA8e\TadA8e-32aa linker-nCas9-spG(D10A)-3NLS
物種來源: Streptococcus pyogenes for Cas9; E. coli for TadA
基因突變: D10A, D1135L, S1136W, G1218K, E1219Q, R1335Q, T1337R for Cas9; W23R, H36L, P48A, R51L, L84F, A106V, D108N, A109S, T111R, D11l9N, H122N, H123Y, S146C, F149Y, R152P, E155V, I156F, K157N, T166I, D167N for TadA
啟動子: maize Ubiquitin-1, OsU3
克隆方法: Gibson Cloning
5’測序引物 N/A

文獻引用方法
材料和方法部分: pH-ABE8e-spG was a gift from Caixia Gao (Addgene plasmid # 200775 ; //n2t.net/addgene:200775 ; RRID:Addgene_200775)
參考文獻部分: Tuning plant phenotypes by precise, graded downregulation of gene expression. Xue C, Qiu F, Wang Y, Li B, Zhao KT, Chen K, Gao C. Nat Biotechnol. 2023 Mar 9. doi: 10.1038/s41587-023-01707-w. 10.1038/s41587-023-01707-w PubMed 36894598

pH-ABE8e-spG質粒圖譜載體圖譜和pH-ABE8e-spG載體序列質粒序列多克隆位點信息

pH-ABE8e-spG載體圖譜

pH-ABE8e-spG質粒載體簡介

pH-ABE8e-spG質粒序列載體序列

LOCUS       pH-ABE8e-spG       17211 bp    DNA     circular SYN 14-APR-2025
DEFINITION  For plant adenine base editing in rice plants or monocotyledons 
            protoplasts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17211)
  AUTHORS   Xue C, Qiu F, Wang Y, Li B, Zhao KT, Chen K, Gao C
  TITLE     Tuning plant phenotypes by precise, graded downregulation of gene 
            expression.
  JOURNAL   Nat Biotechnol. 2023 Mar 9. doi: 10.1038/s41587-023-01707-w.
  PUBMED    36894598
REFERENCE   2  (bases 1 to 17211)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            //gt12365.cn
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2023 Mar 9. doi: 10.1038/s41587-023-01707-w."
COMMENT     Sequence Label: pH-ABE8e-spG
FEATURES             Location/Qualifiers
     source          1..17211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(578..1207)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    2508..2532
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     2720..2742
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2734..2751
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2735..2751
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2818..3197
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     misc_RNA        3233..3308
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3612..5604
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             10328..10348
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10361..10381
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10394..10414
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(11073..11089)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11073..11089)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11086..11108)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11097..11113
                     /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(join(11121..11127,11128..11145,11146..11151))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11166..11187
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        11378..12054
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     11991..12011
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             12121..13146
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    13187..13361
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    13439..13463
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             13888..14682
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      14769..15357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     15258..15277
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     15511..15528
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    15543..15683
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(15609..15628)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     15769..15791
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      16027..16221
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(join(16287..17211,1..149))
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
ORIGIN
        1 tcagcggccg cttgccacga ttcaccgggg cgtgcttctg cctcgatgcg ttgccgctgg
       61 gcggcctgcg cggccttcaa cttctccacc aggtcatcac ccagcgccgc gccgatttgt
      121 accgggccgg atggtttgcg accgctcacg ccgattcctc gggcttgggg gttccagtgc
      181 cattgcaggg ccggcaggca acccagccgc ttacgcctgg ccaaccgccc gttcctccac
      241 acatggggca ttccacggcg tcggtgcctg gttgttcttg attttccatg ccgcctcctt
      301 tagccgctaa aattcatcta ctcatttatt catttgctca tttactctgg tagctgcgcg
      361 atgtattcag atagcagctc ggtaatggtc ttgccttggc gtaccgcgta catcttcagc
      421 ttggtgtgat cctccgccgg caactgaaag ttgacccgct tcatggctgg cgtgtctgcc
      481 aggctggcca acgttgcagc cttgctgctg cgtgcgctcg gacggccggc acttagcgtg
      541 tttgtgcttt tgctcatttt ctctttacct cattaactca aatgagtttt gatttaattt
      601 cagcggccag cgcctggacc tcgcgggcag cgtcgccctc gggttctgat tcaagaacgg
      661 ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg gactcaagaa
      721 tgggcagctc gtacccggcc agcgcctcgg caacctcacc gccgatgcgc gtgcctttga
      781 tcgcccgcga cacgacaaag gccgcttgta gccttccatc cgtgacctca atgcgctgct
      841 taaccagctc caccaggtcg gcggtggccc atatgtcgta agggcttggc tgcaccggaa
      901 tcagcacgaa gtcggctgcc ttgatcgcgg acacagccaa gtccgccgcc tggggcgctc
      961 cgtcgatcac tacgaagtcg cgccggccga tggccttcac gtcgcggtca atcgtcgggc
     1021 ggtcgatgcc gacaacggtt agcggttgat cttcccgcac ggccgcccaa tcgcgggcac
     1081 tgccctgggg atcggaatcg actaacagaa catcggcccc ggcgagttgc agggcgcggg
     1141 ctagatgggt tgcgatggtc gtcttgcctg acccgccttt ctggttaagt acagcgataa
     1201 ccttcatgcg ttccccttgc gtatttgttt atttactcat cgcatcatat acgcagcgac
     1261 cgcatgacgc aagctgtttt actcaaatac acatcacctt tttagacggc ggcgctcggt
     1321 ttcttcagcg gccaagctgg ccggccaggc cgccagcttg gcatcagaca aaccggccag
     1381 gatttcatgc agccgcacgg ttgagacgtg cgcgggcggc tcgaacacgt acccggccgc
     1441 gatcatctcc gcctcgatct cttcggtaat gaaaaacggt tcgtcctggc cgtcctggtg
     1501 cggtttcatg cttgttcctc ttggcgttca ttctcggcgg ccgccagggc gtcggcctcg
     1561 gtcaatgcgt cctcacggaa ggcaccgcgc cgcctggcct cggtgggcgt cacttcctcg
     1621 ctgcgctcaa gtgcgcggta cagggtcgag cgatgcacgc caagcagtgc agccgcctct
     1681 ttcacggtgc ggccttcctg gtcgatcagc tcgcgggcgt gcgcgatctg tgccggggtg
     1741 agggtagggc gggggccaaa cttcacgcct cgggccttgg cggcctcgcg cccgctccgg
     1801 gtgcggtcga tgattaggga acgctcgaac tcggcaatgc cggcgaacac ggtcaacacc
     1861 atgcggccgg ccggcgtggt ggtgtcggcc cacggctctg ccaggctacg caggcccgcg
     1921 ccggcctcct ggatgcgctc ggcaatgtcc agtaggtcgc gggtgctgcg ggccaggcgg
     1981 tctagcctgg tcactgtcac aacgtcgcca gggcgtaggt ggtcaagcat cctggccagc
     2041 tccgggcggt cgcgcctggt gccggtgatc ttctcggaaa acagcttggt gcagccggcc
     2101 gcgtgcagtt cggcccgttg gttggtcaag tcctggtcgt cggtgctgac gcgggcatag
     2161 cccagcaggc cagcggcggc gctcttgttc atggcgtaat gtctccggtt ctagtcgcaa
     2221 gtattctact ttatgcgact aaaacacgcg acaagaaaac gccaggaaaa gggcagggcg
     2281 gcagcctgtc gcgtaactta ggacttgtgc gacatgtcgt tttcagaaga cggctgcact
     2341 gaacgtcaga agccgactgc actatagcag cggaggggtt ggatcaaagt actttgatcc
     2401 cgaggggaac cctgtggttg gcatgcacat acaaatggac gaacggataa accttttcac
     2461 gcccttttaa atatccgatt attctaataa acgctctttt ctcttaggtt tacccgccaa
     2521 tatatcctgt caaacactga tagtttaaac tgaaggcggg aaacgacaat ctgatccaag
     2581 ctcaagctgc tctagcattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     2641 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     2701 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctt
     2761 agtaattcat ccaggtctcc aagttctagg attttcagaa ctgcaactta ttttatcaag
     2821 gaatctttaa acatacgaac agatcactta aagttcttct gaagcaactt aaagttatca
     2881 ggcatgcatg gatcttggag gaatcagatg tgcagtcagg gaccatagca caagacaggc
     2941 gtcttctact ggtgctacca gcaaatgctg gaagccggga acactgggta cgttggaaac
     3001 cacgtgatgt gaagaagtaa gataaactgt aggagaaaag catttcgtag tgggccatga
     3061 agcctttcag gacatgtatt gcagtatggg ccggcccatt acgcaattgg acgacaacaa
     3121 agactagtat tagtaccacc tcggctatcc acatagatca aagctgattt aaaagagttg
     3181 tgcagatgat ccgtggcgaa ccgcaggtga gctgcattaa tcacctgcgc aagttttaga
     3241 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     3301 gtcggtgctt ttttttttcg ttttgcattg agttttctcc gtcgcatgtt tgcagtttta
     3361 ttttccgttt tgcattgaaa tttctccgtc tcatgtttgc agcgtgttca aaaagtacgc
     3421 agctgtattt cacttattta cggcgccaca ttttcatgcc gtttgtgcca actatcccga
     3481 gctagtgaat acagcttggc ttcacacaac actggtgacc cgctgacctg ctcgtacctc
     3541 gtaccgtcgt acggcacagc atttggaatt aaagggtgtg atcgatactg cttgctgcta
     3601 agcttgcatg cctgcagtgc agcgtgaccc ggtcgtgccc ctctctagag ataatgagca
     3661 ttgcatgtct aagttataaa aaattaccac atattttttt tgtcacactt gtttgaagtg
     3721 cagtttatct atctttatac atatatttaa actttactct acgaataata taatctatag
     3781 tactacaata atatcagtgt tttagagaat catataaatg aacagttaga catggtctaa
     3841 aggacaattg agtattttga caacaggact ctacagtttt atctttttag tgtgcatgtg
     3901 ttctcctttt tttttgcaaa tagcttcacc tatataatac ttcatccatt ttattagtac
     3961 atccatttag ggtttagggt taatggtttt tatagactaa tttttttagt acatctattt
     4021 tattctattt tagcctctaa attaagaaaa ctaaaactct attttagttt ttttatttaa
     4081 taatttagat ataaaataga ataaaataaa gtgactaaaa attaaacaaa taccctttaa
     4141 gaaattaaaa aaactaagga aacatttttc ttgtttcgag tagataatgc cagcctgtta
     4201 aacgccgtcg acgagtctaa cggacaccaa ccagcgaacc agcagcgtcg cgtcgggcca
     4261 agcgaagcag acggcacggc atctctgtcg ctgcctctgg acccctctcg agagttccgc
     4321 tccaccgttg gacttgctcc gctgtcggca tccagaaatt gcgtggcgga gcggcagacg
     4381 tgagccggca cggcaggcgg cctcctcctc ctctcacggc accggcagct acgggggatt
     4441 cctttcccac cgctccttcg ctttcccttc ctcgcccgcc gtaataaata gacaccccct
     4501 ccacaccctc tttccccaac ctcgtgttgt tcggagcgca cacacacaca accagatctc
     4561 ccccaaatcc acccgtcggc acctccgctt caaggtacgc cgctcgtcct cccccccccc
     4621 ccctctctac cttctctaga tcggcgttcc ggtccatggt tagggcccgg tagttctact
     4681 tctgttcatg tttgtgttag atccgtgttt gtgttagatc cgtgctgcta gcgttcgtac
     4741 acggatgcga cctgtacgtc agacacgttc tgattgctaa cttgccagtg tttctctttg
     4801 gggaatcctg ggatggctct agccgttccg cagacgggat cgatttcatg attttttttg
     4861 tttcgttgca tagggtttgg tttgcccttt tcctttattt caatatatgc cgtgcacttg
     4921 tttgtcgggt catcttttca tgcttttttt tgtcttggtt gtgatgatgt ggtctggttg
     4981 ggcggtcgtt ctagatcgga gtagaattct gtttcaaact acctggtgga tttattaatt
     5041 ttggatctgt atgtgtgtgc catacatatt catagttacg aattgaagat gatggatgga
     5101 aatatcgatc taggataggt atacatgttg atgcgggttt tactgatgca tatacagaga
     5161 tgctttttgt tcgcttggtt gtgatgatgt ggtgtggttg ggcggtcgtt cattcgttct
     5221 agatcggagt agaatactgt ttcaaactac ctggtgtatt tattaatttt ggaactgtat
     5281 gtgtgtgtca tacatcttca tagttacgag tttaagatgg atggaaatat cgatctagga
     5341 taggtataca tgttgatgtg ggttttactg atgcatatac atgatggcat atgcagcatc
     5401 tattcatatg ctctaacctt gagtacctat ctattataat aaacaagtat gttttataat
     5461 tattttgatc ttgatatact tggatgatgg catatgcagc agctatatgt ggattttttt
     5521 agccctgcct tcatacgcta tttatttgct tggtactgtt tcttttgtcg atgctcaccc
     5581 tgttgtttgg tgttacttct gcagccctag gatgagcgaa gttgaattta gccatgaata
     5641 ttggatgcgc catgcactta cacttgcaaa gagagcccgc gatgaacgcg aagttcctgt
     5701 tggagcagtt cttgttctta acaaccgcgt tatcggagaa ggatggaacc gcgcaatcgg
     5761 acttcatgat cctacagcac atgcagaaat catggcactt cgccagggag gacttgttat
     5821 gcagaactat cgccttatcg atgcaacact ttatgttaca tttgaacctt gcgttatgtg
     5881 cgcaggagca atgatccata gccgcatcgg acgcgttgtg ttcggagtac gtaacagcaa
     5941 acgcggagca gcaggaagcc ttatgaacgt tcttaactat cctggaatga accatcgcgt
     6001 tgaaatcaca gaaggaatcc ttgcagatga atgcgcagca cttctttgcg atttctaccg
     6061 tatgcctcgc caggtgttca atgcgcagaa gaaggcccaa agcagcatca actcaggtgg
     6121 tagctccggc ggctcgtcag gttccgaaac acctggaaca agcgaaagcg caacacctga
     6181 atctagtgga ggaagttctg gtgggtccct taaggacaag aagtactcga tcggcctcgc
     6241 catcgggacg aactcagttg gctgggccgt gatcaccgac gagtacaagg tgccctctaa
     6301 gaagttcaag gtcctgggga acaccgaccg ccattccatc aagaagaacc tcatcggcgc
     6361 tctcctgttc gacagcgggg agaccgctga ggctacgagg ctcaagagaa ccgctaggcg
     6421 ccggtacacg agaaggaaga acaggatctg ctacctccaa gagattttct ccaacgagat
     6481 ggccaaggtt gacgattcat tcttccaccg cctggaggag tctttcctcg tggaggagga
     6541 taagaagcac gagcggcatc ccatcttcgg caacatcgtg gacgaggttg cctaccacga
     6601 gaagtaccct acgatctacc atctgcggaa gaagctcgtg gactccaccg ataaggcgga
     6661 cctcagactg atctacctcg ctctggccca catgatcaag ttccgcggcc atttcctgat
     6721 cgagggggat ctcaacccag acaacagcga tgttgacaag ctgttcatcc aactcgtgca
     6781 gacctacaac caactcttcg aggagaaccc gatcaacgcc tctggcgtgg acgcgaaggc
     6841 tatcctgtcc gcgaggctct cgaagtccag gaggctggag aacctgatcg ctcagctccc
     6901 aggcgagaag aagaacggcc tgttcgggaa cctcatcgct ctcagcctgg ggctcacccc
     6961 gaacttcaag tcgaacttcg atctcgctga ggacgccaag ctgcaactct ccaaggacac
     7021 ctacgacgat gacctcgata acctcctggc ccagatcggc gatcaatacg cggacctgtt
     7081 cctcgctgcc aagaacctgt cggacgccat cctcctgtca gatatcctcc gcgtgaacac
     7141 cgagatcacg aaggctccac tctctgcctc catgatcaag cgctacgacg agcaccatca
     7201 ggatctgacc ctcctgaagg cgctggtccg ccaacagctc ccggagaagt acaaggagat
     7261 tttcttcgat cagtcgaaga acggctacgc tgggtacatc gacggcgggg cctcacaaga
     7321 ggagttctac aagttcatca agccaatcct ggagaagatg gacggcacgg aggagctcct
     7381 ggtgaagctc aacagggagg acctcctgcg gaagcagaga accttcgata acggcagcat
     7441 cccccaccaa atccatctcg gggagctgca cgccatcctg agaaggcaag aggacttcta
     7501 ccctttcctc aaggataacc gggagaagat cgagaagatc ctgaccttca gaatcccata
     7561 ctacgtcggc cctctcgcgc gggggaactc aagattcgct tggatgaccc gcaagtctga
     7621 ggagaccatc acgccgtgga acttcgagga ggtggtggac aagggcgcta gcgctcagtc
     7681 gttcatcgag aggatgacca acttcgacaa gaacctgccc aacgagaagg tgctccctaa
     7741 gcactcgctc ctgtacgagt acttcaccgt ctacaacgag ctcacgaagg tgaagtacgt
     7801 caccgagggc atgcgcaagc cagcgttcct gtccggggag cagaagaagg ctatcgtgga
     7861 cctcctgttc aagaccaacc ggaaggtcac ggttaagcaa ctcaaggagg actacttcaa
     7921 gaagatcgag tgcttcgatt cggtcgagat cagcggcgtt gaggaccgct tcaacgccag
     7981 cctcgggacc taccacgatc tcctgaagat catcaaggat aaggacttcc tggacaacga
     8041 ggagaacgag gatatcctgg aggacatcgt gctgaccctc acgctgttcg aggacaggga
     8101 gatgatcgag gagcgcctga agacgtacgc ccatctcttc gatgacaagg tcatgaagca
     8161 actcaagcgc cggagataca ccggctgggg gaggctgtcc cgcaagctca tcaacggcat
     8221 ccgggacaag cagtccggga agaccatcct cgacttcctc aagagcgatg gcttcgccaa
     8281 caggaacttc atgcaactga tccacgatga cagcctcacc ttcaaggagg atatccaaaa
     8341 ggctcaagtg agcggccagg gggactcgct gcacgagcat atcgcgaacc tcgctggctc
     8401 ccccgcgatc aagaagggca tcctccagac cgtgaaggtt gtggacgagc tcgtgaaggt
     8461 catgggccgg cacaagcctg agaacatcgt catcgagatg gccagagaga accaaaccac
     8521 gcagaagggg caaaagaact ctagggagcg catgaagcgc atcgaggagg gcatcaagga
     8581 gctggggtcc caaatcctca aggagcaccc agtggagaac acccaactgc agaacgagaa
     8641 gctctacctg tactacctcc agaacggcag ggatatgtac gtggaccaag agctggatat
     8701 caaccgcctc agcgattacg acgtcgatca tatcgttccc cagtctttcc tgaaggatga
     8761 ctccatcgac aacaaggtcc tcaccaggtc ggacaagaac cgcggcaagt cagataacgt
     8821 tccatctgag gaggtcgtta agaagatgaa gaactactgg aggcagctcc tgaacgccaa
     8881 gctgatcacg caaaggaagt tcgacaacct caccaaggct gagagaggcg ggctctcaga
     8941 gctggacaag gccggcttca tcaagcggca gctggtcgag accagacaaa tcacgaagca
     9001 cgttgcgcaa atcctcgact ctcggatgaa cacgaagtac gatgagaacg acaagctgat
     9061 cagggaggtt aaggtgatca ccctgaagtc taagctcgtc tccgacttca ggaaggattt
     9121 ccagttctac aaggttcgcg agatcaacaa ctaccaccat gcccatgacg cttacctcaa
     9181 cgctgtggtc ggcaccgctc tgatcaagaa gtacccaaag ctggagtccg agttcgtgta
     9241 cggggactac aaggtttacg atgtgcgcaa gatgatcgcc aagtcggagc aagagatcgg
     9301 caaggctacc gccaagtact tcttctactc aaacatcatg aacttcttca agaccgagat
     9361 cacgctggcc aacggcgaga tccggaagag accgctcatc gagaccaacg gcgagacggg
     9421 ggagatcgtg tgggacaagg gcagggattt cgcgaccgtc cgcaaggttc tctccatgcc
     9481 ccaggtgaac atcgtcaaga agaccgaggt ccaaacgggc gggttctcaa aggagtctat
     9541 cctgcctaag cggaacagcg acaagctcat cgccagaaag aaggactggg acccaaagaa
     9601 gtacggcggg ttcctctggc ctaccgtggc ctactcggtc ctggttgtgg cgaaggttga
     9661 gaagggcaag tccaagaagc tcaagagcgt gaaggagctc ctggggatca ccatcatgga
     9721 gaggtccagc ttcgagaaga acccaatcga cttcctggag gccaagggct acaaggaggt
     9781 gaagaaggac ctgatcatca agctcccgaa gtactctctc ttcgagctgg agaacggcag
     9841 gaagagaatg ctggcttccg ctaagcagct ccagaagggg aacgagctcg cgctgccaag
     9901 caagtacgtg aacttcctct acctggcttc ccactacgag aagctcaagg gcagcccgga
     9961 ggacaacgag caaaagcagc tgttcgtcga gcagcacaag cattacctcg acgagatcat
    10021 cgagcaaatc tccgagttca gcaagcgcgt gatcctcgcc gacgcgaacc tggataaggt
    10081 cctctccgcc tacaacaagc accgggacaa gcccatcaga gagcaagcgg agaacatcat
    10141 ccatctcttc accctgacga acctcggcgc tcctgctgct ttcaagtact tcgacaccac
    10201 gatcgatcgg aagcaataca ggtccacgaa ggaggtcctg gacgcgaccc tcatccacca
    10261 gtcgatcacc ggcctgtacg agacgaggat cgacctctca caactcggcg gggattccgg
    10321 cggcagccca aagaagaagc ggaaggtgtc tggaggttct cctaagaaaa agagaaaagt
    10381 gtccggcggc tccccgaaga agaagcgcaa ggtgtaggag ctcagagctt tcgttcgtat
    10441 catcggtttc gacaacgttc gtcaagttca atgcatcagt ttcattgcgc acacaccaga
    10501 atcctactga gtttgagtat tatggcattg ggaaaactgt ttttcttgta ccatttgttg
    10561 tgcttgtaat ttactgtgtt ttttattcgg ttttcgctat cgaactgtga aatggaaatg
    10621 gatggagaag agttaatgaa tgatatggtc cttttgttca ttctcaaatt aatattattt
    10681 gttttttctc ttatttgttg tgtgttgaat ttgaaattat aagagatatg caaacatttt
    10741 gttttgagta aaaatgtgtc aaatcgtggc ctctaatgac cgaagttaat atgaggagta
    10801 aaacacttgt agttgtacca ttatgcttat tcactaggca acaaatatat tttcagacct
    10861 agaaaagctg caaatgttac tgaatacaag tatgtcctct tgtgttttag acatttatga
    10921 actttccttt atgtaatttt ccagaatcct tgtcagattc taatcattgc tttataatta
    10981 tagttatact catggatttg tagttgagta tgaaaatatt ttttaatgca ttttatgact
    11041 tgccaattga ttgacaacga attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt
    11101 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
    11161 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
    11221 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
    11281 cgtattggct agagcagctt gccaacatgg tggagcacga cactctcgtc tactccaaga
    11341 atatcaaaga tacagtctca gaagaccaaa gggctattga gacttttcaa caaagggtaa
    11401 tatcgggaaa cctcctcgga ttccattgcc cagctatctg tcacttcatc aaaaggacag
    11461 tagaaaagga aggtggcacc tacaaatgcc atcattgcga taaaggaaag gctatcgttc
    11521 aagatgcctc tgccgacagt ggtcccaaag atggaccccc acccacgagg agcatcgtgg
    11581 aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga ttgatgtgaa catggtggag
    11641 cacgacactc tcgtctactc caagaatatc aaagatacag tctcagaaga ccaaagggct
    11701 attgagactt ttcaacaaag ggtaatatcg ggaaacctcc tcggattcca ttgcccagct
    11761 atctgtcact tcatcaaaag gacagtagaa aaggaaggtg gcacctacaa atgccatcat
    11821 tgcgataaag gaaaggctat cgttcaagat gcctctgccg acagtggtcc caaagatgga
    11881 cccccaccca cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa
    11941 gtggattgat gtgatatctc cactgacgta agggatgacg cacaatccca ctatccttcg
    12001 caagaccctt cctctatata aggaagttca tttcatttgg agaggacacg ctgaaatcac
    12061 cagtctctct ctacaaatct atctctctcg agctttcgca gatccggggg gcaatgagat
    12121 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
    12181 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
    12241 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
    12301 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
    12361 ggggagttta gcgagagcct gacctattgc atctcccgcc gttcacaggg tgtcacgttg
    12421 caagacctgc ctgaaaccga actgcccgct gttctacaac cggtcgcgga ggctatggat
    12481 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
    12541 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
    12601 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
    12661 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
    12721 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
    12781 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
    12841 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgcca
    12901 cgactccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
    12961 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
    13021 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
    13081 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
    13141 aaatagagta gatgccgacc gggatctgtc gatcgacaag ctcgagtttc tccataataa
    13201 tgtgtgagta gttcccagat aagggaatta gggttcctat agggtttcgc tcatgtgttg
    13261 agcatataag aaacccttag tatgtatttg tatttgtaaa atacttctat caataaaatt
    13321 tctaattcct aaaaccaaaa tccagtacta aaatccagat cccccgaatt aattcggcgt
    13381 taattcagta cattaaaaac gtccgcaatg tgttattaag ttgtctaagc gtcaatttgt
    13441 ttacaccaca atatatcctg ccaccagcca gccaacagct ccccgaccgg cagctcggca
    13501 caaaatcacc actcgataca ggcagcccat cagtccggga cggcgtcagc gggagagccg
    13561 ttgtaaggcg gcagactttg ctcatgttac cgatgctatt cggaagaacg gcaactaagc
    13621 tgccgggttt gaaacacgga tgatctcgcg gagggtagca tgttgattgt aacgatgaca
    13681 gagcgttgct gcctgtgatc accgcggttt caaaatcggc tccgtcgata ctatgttata
    13741 cgccaacttt gaaaacaact ttgaaaaagc tgttttctgg tatttaaggt tttagaatgc
    13801 aaggaacagt gaattggagt tcgtcttgtt ataattagct tcttggggta tctttaaata
    13861 ctgtagaaaa gaggaaggaa ataataaatg gctaaaatga gaatatcacc ggaattgaaa
    13921 aaactgatcg aaaaataccg ctgcgtaaaa gatacggaag gaatgtctcc tgctaaggta
    13981 tataagctgg tgggagaaaa tgaaaaccta tatttaaaaa tgacggacag ccggtataaa
    14041 gggaccacct atgatgtgga acgggaaaag gacatgatgc tatggctgga aggaaagctg
    14101 cctgttccaa aggtcctgca ctttgaacgg catgatggct ggagcaatct gctcatgagt
    14161 gaggccgatg gcgtcctttg ctcggaagag tatgaagatg aacaaagccc tgaaaagatt
    14221 atcgagctgt atgcggagtg catcaggctc tttcactcca tcgacatatc ggattgtccc
    14281 tatacgaata gcttagacag ccgcttagcc gaattggatt acttactgaa taacgatctg
    14341 gccgatgtgg attgcgaaaa ctgggaagaa gacactccat ttaaagatcc gcgcgagctg
    14401 tatgattttt taaagacgga aaagcccgaa gaggaacttg tcttttccca cggcgacctg
    14461 ggagacagca acatctttgt gaaagatggc aaagtaagtg gctttattga tcttgggaga
    14521 agcggcaggg cggacaagtg gtatgacatt gccttctgcg tccggtcgat cagggaggat
    14581 atcggggaag aacagtatgt cgagctattt tttgacttac tggggatcaa gcctgattgg
    14641 gagaaaataa aatattatat tttactggat gaattgtttt agtacctaga atgcatgacc
    14701 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
    14761 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
    14821 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
    14881 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
    14941 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
    15001 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
    15061 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
    15121 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
    15181 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
    15241 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
    15301 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
    15361 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
    15421 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
    15481 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
    15541 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
    15601 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
    15661 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
    15721 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
    15781 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagggtgcc ttgatgtggg
    15841 cgccggcggt cgagtggcga cggcgcggct tgtccgcgcc ctggtagatt gcctggccgt
    15901 aggccagcca tttttgagcg gccagcggcc gcgataggcc gacgcgaagc ggcggggcgt
    15961 agggagcgca gcgaccgaag ggtaggcgct ttttgcagct cttcggctgt gcgctggcca
    16021 gacagttatg cacaggccag gcgggtttta agagttttaa taagttttaa agagttttag
    16081 gcggaaaaat cgcctttttt ctcttttata tcagtcactt acatgtgtga ccggttccca
    16141 atgtacggct ttgggttccc aatgtacggg ttccggttcc caatgtacgg ctttgggttc
    16201 ccaatgtacg tgctatccac aggaaacaga ccttttcgac ctttttcccc tgctagggca
    16261 atttgcccta gcatctgctc cgtacattag gaaccggcgg atgcttcgcc ctcgatcagg
    16321 ttgcggtagc gcatgactag gatcgggcca gcctgccccg cctcctcctt caaatcgtac
    16381 tccggcaggt catttgaccc gatcagcttg cgcacggtga aacagaactt cttgaactct
    16441 ccggcgctgc cactgcgttc gtagatcgtc ttgaacaacc atctggcttc tgccttgcct
    16501 gcggcgcggc gtgccaggcg gtagagaaaa cggccgatgc cgggatcgat caaaaagtaa
    16561 tcggggtgaa ccgtcagcac gtccgggttc ttgccttctg tgatctcgcg gtacatccaa
    16621 tcagctagct cgatctcgat gtactccggc cgcccggttt cgctctttac gatcttgtag
    16681 cggctaatca aggcttcacc ctcggatacc gtcaccaggc ggccgttctt ggccttcttc
    16741 gtacgctgca tggcaacgtg cgtggtgttt aaccgaatgc aggtttctac caggtcgtct
    16801 ttctgctttc cgccatcggc tcgccggcag aacttgagta cgtccgcaac gtgtggacgg
    16861 aacacgcggc cgggcttgtc tcccttccct tcccggtatc ggttcatgga ttcggttaga
    16921 tgggaaaccg ccatcagtac caggtcgtaa tcccacacac tggccatgcc ggccggccct
    16981 gcggaaacct ctacgtgccc gtctggaagc tcgtagcgga tcacctcgcc agctcgtcgg
    17041 tcacgcttcg acagacggaa aacggccacg tccatgatgc tgcgactatc gcgggtgccc
    17101 acgtcataga gcatcggaac gaaaaaatct ggttgctcgt cgcccttggg cggcttccta
    17161 atcgacggcg caccggctgc cggcggttgc cgggattctt tgcggattcg a
//

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