ABE8.8-m

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联系方式:I47-825O-882O

相关技术服务:质粒构建    基因合成    质粒大提

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载体基本信息

质粒名称: ABE8.8-m
出品公司: addgene
目录编号: 136294
存储实验室: Nicole Gaudelli

载体骨架基本信息
载体名称: pBR322
载体出品公司:
空载体大小: 3400 bp
完整质粒大小: 8166 bp
载体类型: 哺乳细胞表达
载体抗性: 氨苄霉素
生长温度: 37 ℃
宿主菌: NEB Stable
拷贝数: 高拷贝

插入基因信息
插入基因名称: ABE8.8-m
插入大小: 5300 bp
基因突变: D10A in S. pyogenes Cas9, TadA mutations described in manuscript
标签/融合蛋白: C-terminal BPNLS (C terminal on insert)
克隆方法: Ligation Independent Cloning
5’测序引物 T7
3’测序引物 M13R

文献引用方法
材料和方法部分: ABE8.8-m was a gift from Nicole Gaudelli (Addgene plasmid # 136294 ; http://n2t.net/addgene:136294 ; RRID:Addgene_136294)
参考文献部分:
 
Directed evolution of adenine base editors with increased activity and therapeutic application. Gaudelli NM, Lam DK, Rees HA, Sola-Esteves NM, Barrera LA, Born DA, Edwards A, Gehrke JM, Lee SJ, Liquori AJ, Murray R, Packer MS, Rinaldi C, Slaymaker IM, Yen J, Young LE, Ciaramella G. Nat Biotechnol. 2020 Apr 13. pii: 10.1038/s41587-020-0491-6. doi: 10.1038/s41587-020-0491-6. 10.1038/s41587-020-0491-6 PubMed 32284586

载体图谱质粒图谱和多克隆位点信息

ABE8.8-m载体图谱

载体简介

载体序列

LOCUS       ABE8.8-m                8166 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  expresses ABE8.8-m in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    ABE8.8-m
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8166)
  AUTHORS   Gaudelli NM, Lam DK, Rees HA, Sola-Esteves NM, Barrera LA, Born DA, 
            Edwards A, Gehrke JM, Lee SJ, Liquori AJ, Murray R, Packer MS, 
            Rinaldi C, Slaymaker IM, Yen J, Young LE, Ciaramella G
  TITLE     Directed evolution of adenine base editors with increased activity 
            and therapeutic application.
  JOURNAL   Nat Biotechnol. 2020 Apr 13. pii: 10.1038/s41587-020-0491-6. doi: 
            10.1038/s41587-020-0491-6.
  PUBMED    32284586
REFERENCE   2  (bases 1 to 8166)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8166
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        111..490
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        491..694
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     645..665
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     736..755
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        736..754
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             771..1268
                     /codon_start=1
                     /product="DNA adenosine deaminase evolved from the E. coli 
                     tRNA adenosine deaminase TadA (Gaudelli et al., 2017)"
                     /label=TadA*(7.10)
                     /translation="SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWN
                     RAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFG
                     VRNAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLCRFFRMPRRVFNAQKKAQS
                     STD"
     CDS             1365..5465
                     /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"
     CDS             5511..5531
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5540..5557
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5580..5597)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5586..5810
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5881..5897)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5881..5897)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5894..5916)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5905..5921
                     /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(5929..5959)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5974..5995
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6112..6129)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6283..6871)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6363..6382)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7042..7902)
                     /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     7665..7684
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7903..8007)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8086..8105)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
       61 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
      121 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      181 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      241 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      301 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      361 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      421 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      481 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      541 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      601 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      661 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      721 gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg tccgaagtcg
      781 agttttccca tgagtactgg atgagacacg cattgactct cgcaaagagg gctcgagatg
      841 aacgcgaggt gcccgtgggg gcagtactcg tgctcaacaa tcgcgtaatc ggcgaaggtt
      901 ggaatagggc aatcggactc cacgacccca ctgcacatgc ggaaatcatg gcccttcgac
      961 agggagggct tgtgatgcag aattatcgac ttatcgatgc gacgctgtac gtcacgtttg
     1021 aaccttgcgt aatgtgcgcg ggagctatga ttcactcccg cattggacga gttgtattcg
     1081 gtgttcgcaa cgccaagacg ggtgccgcag gttcactgat ggacgtgctg catcatccag
     1141 gcatgaacca ccgggtagaa atcacagaag gcatattggc ggacgaatgt gcggcgctgt
     1201 tgtgtcgttt ttttcgcatg cccaggcggg tctttaacgc ccagaaaaaa gcacaatcct
     1261 ctactgactc tggtggttct tctggtggtt ctagcggcag cgagactccc gggacctcag
     1321 agtccgccac acccgaaagt tctggtggtt cttctggtgg ttctgacaag aagtacagca
     1381 tcggcctggc catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
     1441 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
     1501 tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa
     1561 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
     1621 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
     1681 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
     1741 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
     1801 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
     1861 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
     1921 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
     1981 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
     2041 cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg
     2101 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
     2161 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
     2221 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
     2281 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
     2341 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
     2401 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag
     2461 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
     2521 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
     2581 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
     2641 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
     2701 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
     2761 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt
     2821 ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
     2881 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag
     2941 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg
     3001 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
     3061 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
     3121 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
     3181 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
     3241 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
     3301 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
     3361 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
     3421 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
     3481 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
     3541 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
     3601 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
     3661 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
     3721 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
     3781 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
     3841 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc
     3901 tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga
     3961 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc
     4021 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg
     4081 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
     4141 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg
     4201 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
     4261 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
     4321 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
     4381 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
     4441 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
     4501 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
     4561 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc
     4621 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
     4681 aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg
     4741 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
     4801 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
     4861 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
     4921 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
     4981 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
     5041 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
     5101 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg
     5161 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
     5221 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg
     5281 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
     5341 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc
     5401 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
     5461 gtgacgaggg agctgataag cgcaccgccg atggttccga gttcgaaagc cccaagaaga
     5521 agaggaaagt ctaaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     5581 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     5641 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     5701 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     5761 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     5821 ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg cgtaatcatg
     5881 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5941 cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca cattaattgc
     6001 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6061 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6121 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6181 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6241 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6301 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6361 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6421 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6481 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6541 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6601 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6661 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6721 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6781 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6841 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6901 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6961 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7021 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7081 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7141 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7201 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7261 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7321 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7381 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7441 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7501 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7561 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7621 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7681 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7741 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7801 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7861 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7921 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7981 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     8041 cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa tctgctctga
     8101 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     8161 cgcgag
//

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