PABE-1

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PABE1载体基本信息

质粒名称: PABE-1 
出品公司: 爱迪基因
目录编号: 115622
存储实验室: Caixia Gao
载体骨架基本信息
载体名称: pJIT163
载体出品公司: --
载体抗性: 氨苄青霉素
空载体大小: --
完整质粒大小: --
载体修饰: --
载体类型: 植物表达,CRISPR
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: NLS-wtTadA-TadA7.10-nCas9-NLS
物种来源: --
插入大小: --
基因突变: --
GenBank编号 --
启动子 --
标签/融合蛋白: --
克隆方法: 限制性内切酶
5' 克隆位点: --
3' 克隆位点: --
5' 测序引物 --
3' 测序引物 --

文献引用方法
材料和方法部分: PABE-1 was a gift from Caixia Gao (Addgene plasmid # 115622 ; http://n2t.net/addgene:115622 ; RRID:Addgene_115622)
参考文献部分: Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion. Li C, Zong Y, Wang Y, Jin S, Zhang D, Song Q, Zhang R, Gao C. Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z. 10.1186/s13059-018-1443-z PubMed 29807545

PABE1载体图谱质粒图谱和多克隆位点信息

PABE-1质粒图谱

PABE1载体简介

PABE1载体序列

LOCUS       PABE-1               10342 bp ds-DNA     circular SYN 10-JAN-2019
DEFINITION  Targeted A to G in wheat plants or monocotyledons protoplasts .
ACCESSION   .
VERSION     .
KEYWORDS    PABE-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10342)
  AUTHORS   Li C, Zong Y, Wang Y, Jin S, Zhang D, Song Q, Zhang R, Gao C
  TITLE     Expanded base editing in rice and wheat using a Cas9-adenosine 
            deaminase fusion.
  JOURNAL   Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z.
  PUBMED    29807545
REFERENCE   2  (bases 1 to 10342)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 10, 2019 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10342
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             814..834
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2041..6141
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             6054..6062
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             6142..6189
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    6247..6423
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(6977..6993)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6977..6993)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6990..7012)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7001..7017
                     /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(7025..7055)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7070..7091
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7208..7225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7379..7967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7459..7478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8138..8998)
                     /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     8761..8780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8999..9103)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatcctggga tggctctagc cgttccgcag acgggatcga tttcatgatt ttttttgttt
       61 cgttgcatag ggtttggttt gcccttttcc tttatttcaa tatatgccgt gcacttgttt
      121 gtcgggtcat cttttcatgc ttttttttgt cttggttgtg atgatgtggt ctggttgggc
      181 ggtcgttcta gatcggagta gaattaattc tgtttcaaac tacctggtgg atttattaat
      241 tttggatctg tatgtgtgtg ccatacatat tcatagttac gaattgaaga tgatggatgg
      301 aaatatcgat ctaggatagg tatacatgtt gatgcgggtt ttactgatgc atatacagag
      361 atgctttttg ttcgcttggt tgtgatgatg tggtgtggtt gggcggtcgt tcattcgttc
      421 tagatcggag tagaatactg tttcaaacta cctggtgtat ttattaattt tggaactgta
      481 tgtgtgtgtc atacatcttc atagttacga gtttaagatg gatggaaata tcgatctagg
      541 ataggtatac atgttgatgt gggttttact gatgcatata catgatggca tatgcagcat
      601 ctattcatat gctctaacct tgagtaccta tctattataa taaacaagta tgttttataa
      661 ttattttgat cttgatatac ttggatgatg gcatatgcag cagctatatg tggatttttt
      721 tagccctgcc ttcatacgct atttatttgc ttggtactgt ttcttttgtc gatgctcacc
      781 ctgttgtttg gtgttacttc tgcaaagctt atgccaaaaa agaagagaaa ggtttcaggc
      841 ggctcctccg aggtggagtt ctctcacgag tattggatga ggcacgctct tacacttgct
      901 aagagagctt gggacgaaag agaagtgcca gttggcgccg ttcttgtgca taataatagg
      961 gtgatcggcg agggttggaa tagaccaatt ggaaggcatg atccaacagc tcacgcagag
     1021 attatggctc tcagacaagg cggcctcgtt atgcagaact acaggctcat tgacgctaca
     1081 ctctacgtga cactcgaacc ttgcgttatg tgcgccggag ctatgattca ttctaggatt
     1141 ggcagggtcg tgtttggagc tagggacgct aaaacaggag ccgccggatc tcttatggac
     1201 gtgttgcatc atccaggcat gaaccatagg gtggagatta cagagggcat tcttgcagac
     1261 gagtgcgctg ctcttctttc cgatttcttc aggatgagaa ggcaggagat taaggcccag
     1321 aagaaggctc agtcttctac agatagcgga ggatcttccg gaggatctag cggctccgag
     1381 acaccaggaa catccgaaag cgctacacca gaatctagcg gaggctcttc cggaggatct
     1441 tctgaagtgg agttctccca cgagtattgg atgaggcacg ctcttacact tgctaaaagg
     1501 gctagggacg aaagggaagt tccagttgga gctgttctcg tgctcaataa cagggtgatt
     1561 ggcgagggtt ggaatagagc cattggactc catgatccaa cagctcacgc agagattatg
     1621 gctcttagac aaggcggcct cgttatgcag aattacagac tcatcgacgc cacactctac
     1681 gttaccttcg aaccttgcgt tatgtgcgcc ggagctatga tccattctag gattggcagg
     1741 gtcgtgttcg gcgttagaaa cgctaagaca ggagctgcag gctctcttat ggacgttctt
     1801 cattacccag gcatgaatca tagagtggag atcacagaag gcattcttgc agacgagtgc
     1861 gcagctctcc tttgctattt cttcaggatg ccgaggcaag ttttcaacgc tcagaagaag
     1921 gcccagtctt ctacagattc cggcggatct tctggaggat ctagcggctc cgagacacca
     1981 ggaacatccg aatccgctac accagagtct tctggaggat ctagcggagg atctcttaag
     2041 gacaagaagt actcgatcgg cctcgccatt gggactaact ctgttggctg ggccgtgatc
     2101 accgacgagt acaaggtgcc ctcaaagaag ttcaaggtcc tgggcaacac cgatcggcat
     2161 tccatcaaga agaatctcat tggcgctctc ctgttcgaca gcggcgagac ggctgaggct
     2221 acgcggctca agcgcaccgc ccgcaggcgg tacacgcgca ggaagaatcg catctgctac
     2281 ctgcaggaga ttttctccaa cgagatggcg aaggttgacg attctttctt ccacaggctg
     2341 gaggagtcat tcctcgtgga ggaggataag aagcacgagc ggcatccaat cttcggcaac
     2401 attgtcgacg aggttgccta ccacgagaag taccctacga tctaccatct gcggaagaag
     2461 ctcgtggact ccacagataa ggcggacctc cgcctgatct acctcgctct ggcccacatg
     2521 attaagttca ggggccattt cctgatcgag ggggatctca acccggacaa tagcgatgtt
     2581 gacaagctgt tcatccagct cgtgcagacg tacaaccagc tcttcgagga gaaccccatt
     2641 aatgcgtcag gcgtcgacgc gaaggctatc ctgtccgcta ggctctcgaa gtctcggcgc
     2701 ctcgagaacc tgatcgccca gctgccgggc gagaagaaga acggcctgtt cgggaatctc
     2761 attgcgctca gcctggggct cacgcccaac ttcaagtcga atttcgatct cgctgaggac
     2821 gccaagctgc agctctccaa ggacacatac gacgatgacc tggataacct cctggcccag
     2881 atcggcgatc agtacgcgga cctgttcctc gctgccaaga atctgtcgga cgccatcctc
     2941 ctgtctgata ttctcagggt gaacaccgag attacgaagg ctccgctctc agcctccatg
     3001 atcaagcgct acgacgagca ccatcaggat ctgaccctcc tgaaggcgct ggtcaggcag
     3061 cagctccccg agaagtacaa ggagatcttc ttcgatcagt cgaagaacgg ctacgctggg
     3121 tacattgacg gcggggcctc tcaggaggag ttctacaagt tcatcaagcc gattctggag
     3181 aagatggacg gcacggagga gctgctggtg aagctcaatc gcgaggacct cctgaggaag
     3241 cagcggacat tcgataacgg cagcatccca caccagattc atctcgggga gctgcacgct
     3301 atcctgagga ggcaggagga cttctaccct ttcctcaagg ataaccgcga gaagatcgag
     3361 aagattctga ctttcaggat cccgtactac gtcggcccac tcgctagggg caactcccgc
     3421 ttcgcttgga tgacccgcaa gtcagaggag acgatcacgc cgtggaactt cgaggaggtg
     3481 gtcgacaagg gcgctagcgc tcagtcgttc atcgagagga tgacgaattt cgacaagaac
     3541 ctgccaaatg agaaggtgct ccctaagcac tcgctcctgt acgagtactt cacagtctac
     3601 aacgagctga ctaaggtgaa gtatgtgacc gagggcatga ggaagccggc tttcctgtct
     3661 ggggagcaga agaaggccat cgtggacctc ctgttcaaga ccaaccggaa ggtcacggtt
     3721 aagcagctca aggaggacta cttcaagaag attgagtgct tcgattcggt cgagatctct
     3781 ggcgttgagg accgcttcaa cgcctccctg gggacctacc acgatctcct gaagatcatt
     3841 aaggataagg acttcctgga caacgaggag aatgaggata tcctcgagga cattgtgctg
     3901 acactcactc tgttcgagga ccgggagatg atcgaggagc gcctgaagac ttacgcccat
     3961 ctcttcgatg acaaggtcat gaagcagctc aagaggagga ggtacaccgg ctgggggagg
     4021 ctgagcagga agctcatcaa cggcattcgg gacaagcagt ccgggaagac gatcctcgac
     4081 ttcctgaaga gcgatggctt cgcgaaccgc aatttcatgc agctgattca cgatgacagc
     4141 ctcacattca aggaggatat ccagaaggct caggtgagcg gccaggggga ctcgctgcac
     4201 gagcatatcg cgaacctcgc tggctcgcca gctatcaaga aggggattct gcagaccgtg
     4261 aaggttgtgg acgagctggt gaaggtcatg ggcaggcaca agcctgagaa catcgtcatt
     4321 gagatggccc gggagaatca gaccacgcag aagggccaga agaactcacg cgagaggatg
     4381 aagaggatcg aggagggcat taaggagctg gggtcccaga tcctcaagga gcacccggtg
     4441 gagaacacgc agctgcagaa tgagaagctc tacctgtact acctccagaa tggccgcgat
     4501 atgtatgtgg accaggagct ggatattaac aggctcagcg attacgacgt cgatcatatc
     4561 gttccacagt cattcctgaa ggatgactcc attgacaaca aggtcctcac caggtcggac
     4621 aagaaccggg gcaagtctga taatgttcct tcagaggagg tcgttaagaa gatgaagaac
     4681 tactggcgcc agctcctgaa tgccaagctg atcacgcagc ggaagttcga taacctcaca
     4741 aaggctgaga ggggcgggct ctctgagctg gacaaggcgg gcttcatcaa gaggcagctg
     4801 gtcgagacac ggcagatcac taagcacgtt gcgcagattc tcgactcacg gatgaacact
     4861 aagtacgatg agaatgacaa gctgatccgc gaggtgaagg tcatcaccct gaagtcaaag
     4921 ctcgtctccg acttcaggaa ggatttccag ttctacaagg ttcgggagat caacaattac
     4981 caccatgccc atgacgcgta cctgaacgcg gtggtcggca cagctctgat caagaagtac
     5041 ccaaagctcg agagcgagtt cgtgtacggg gactacaagg tttacgatgt gaggaagatg
     5101 atcgccaagt cggagcagga gattggcaag gctaccgcca agtacttctt ctactctaac
     5161 attatgaatt tcttcaagac agagatcact ctggccaatg gcgagatccg gaagcgcccc
     5221 ctcatcgaga cgaacggcga gacgggggag atcgtgtggg acaagggcag ggatttcgcg
     5281 accgtcagga aggttctctc catgccacaa gtgaatatcg tcaagaagac agaggtccag
     5341 actggcgggt tctctaagga gtcaattctg cctaagcgga acagcgacaa gctcatcgcc
     5401 cgcaagaagg actgggatcc gaagaagtac ggcgggttcg acagccccac tgtggcctac
     5461 tcggtcctgg ttgtggcgaa ggttgagaag ggcaagtcca agaagctcaa gagcgtgaag
     5521 gagctgctgg ggatcacgat tatggagcgc tccagcttcg agaagaaccc gatcgatttc
     5581 ctggaggcga agggctacaa ggaggtgaag aaggacctga tcattaagct ccccaagtac
     5641 tcactcttcg agctggagaa cggcaggaag cggatgctgg cttccgctgg cgagctgcag
     5701 aaggggaacg agctggctct gccgtccaag tatgtgaact tcctctacct ggcctcccac
     5761 tacgagaagc tcaagggcag ccccgaggac aacgagcaga agcagctgtt cgtcgagcag
     5821 cacaagcatt acctcgacga gatcattgag cagatttccg agttctccaa gcgcgtgatc
     5881 ctggccgacg cgaatctgga taaggtcctc tccgcgtaca acaagcaccg cgacaagcca
     5941 atcagggagc aggctgagaa tatcattcat ctcttcaccc tgacgaacct cggcgcccct
     6001 gctgctttca agtacttcga cacaactatc gatcgcaaga ggtacacaag cactaaggag
     6061 gtcctggacg cgaccctcat ccaccagtcg attaccggcc tctacgagac gcgcatcgac
     6121 ctgtctcagc tcgggggcga caagcggcca gcggcgacga agaaggcggg gcaggcgaag
     6181 aagaagaagt aggaattcgg tacgctgaaa tcaccagtct ctctctacaa atctatctct
     6241 ctctattttc tccataaata atgtgtgagt agtttcccga taagggaaat tagggttctt
     6301 atagggtttc gctcatgtgt tgagcatata agaaaccctt agtatgtatt tgtatttgta
     6361 aaatacttct atcaataaaa tttctaattc ctaaaaccaa aatccagtac taaaatccag
     6421 atctcctaaa gtccctatag atctttgtcg tgaatataaa ccagacacga gacgactaaa
     6481 cctggagccc agacgccgtt cgaagctaga agtaccgctt aggcaggagg ccgttaggga
     6541 aaagatgcta aggcagggtt ggttacgttg actcccccgt aggtttggtt taaatatgat
     6601 gaagtggacg gaaggaagga ggaagacaag gaaggataag gttgcaggcc ctgtgcaagg
     6661 taagaagatg gaaatttgat agaggtacgc tactatactt atactatacg ctaagggaat
     6721 gcttgtattt ataccctata ccccctaata accccttatc aatttaagaa ataatccgca
     6781 taagcccccg cttaaaaatt ggtatcagag ccatgaatag gtctatgacc aaaactcaag
     6841 aggataaaac ctcaccaaaa tacgaaagag ttcttaactc taaagataaa agatctttca
     6901 agatcaaaac tagttccctc acaccggtga cggggatcgc atgcgatatc tcgagatcta
     6961 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     7021 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     7081 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     7141 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     7201 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7261 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7321 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7381 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7441 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7501 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7561 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7621 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7681 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7741 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7801 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7861 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7921 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7981 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     8041 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     8101 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     8161 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     8221 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     8281 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     8341 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     8401 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     8461 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     8521 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     8581 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     8641 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     8701 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     8761 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8821 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8881 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8941 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     9001 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     9061 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     9121 tgccacctgc cagtgccaag ctaattcgag ctcggtacct gacccggtcg tgcccctctc
     9181 tagagataat gagcattgca tgtctaagtt ataaaaaatt accacatatt ttttttgtca
     9241 cacttgtttg aagtgcagtt tatctatctt tatacatata tttaaacttt actctacgaa
     9301 taatataatc tatagtacta caataatatc agtgttttag agaatcatat aaatgaacag
     9361 ttagacatgg tctaaaggac aattgagtat tttgacaaca ggactctaca gttttatctt
     9421 tttagtgtgc atgtgttctc cttttttttt gcaaatagct tcacctatat aatacttcat
     9481 ccattttatt agtacatcca tttagggttt agggttaatg gtttttatag actaattttt
     9541 ttagtacatc tattttattc tattttagcc tctaaattaa gaaaactaaa actctatttt
     9601 agttttttta tttaataatt tagatataaa atagaataaa ataaagtgac taaaaattaa
     9661 acaaataccc tttaagaaat taaaaaaact aaggaaacat ttttcttgtt tcgagtagat
     9721 aatgccagcc tgttaaacgc cgtcgacgag tctaacggac accaaccagc gaaccagcag
     9781 cgtcgcgtcg ggccaagcga agcagacggc acggcatctc tgtcgctgcc tctggacccc
     9841 tctcgatcga gagttccgct ccaccgttgg acttgctccg ctgtcggcat ccagaaattg
     9901 cgtggcggag cggcagacgt gagccggcac ggcaggcggc ctcctcctcc tctcacggca
     9961 ccggcagcta cgggggattc ctttcccacc gctccttcgc tttcccttcc tcgcccgccg
    10021 taataaatag acaccccctc cacaccctct ttccccaacc tcgtgttgtt cggagcgcac
    10081 acacacacaa ccagatctcc cccaaatcca cccgtcggca cctccgcttc aaggtacgcc
    10141 gctcgtcctc cccccccccc cctctctacc ttctctagat cggcgttccg gtccatggtt
    10201 agggcccggt agttctactt ctgttcatgt ttgtgttaga tccgtgtttg tgttagatcc
    10261 gtgctgctag cgttcgtaca cggatgcgac ctgtacgtca gacacgttct gattgctaac
    10321 ttgccagtgt ttctctttgg gg
//

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