pCMV-hA3G-BE3

价格:1800元

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: pCMV-hA3G-BE3
出品公司: Addgene
目录编号: 113415
存储实验室: Jia Chen

载体骨架基本信息
载体名称: CMV
空载体大小: 3399 bp
完整质粒大小: 8997bp
载体类型: 哺乳细胞表达、CRISPR
载体抗性: 氨苄霉素
生长温度: 37 ℃
宿主菌: DH5a
拷贝数: 高拷贝

插入基因信息
插入基因名称: hA3G-BE3  /  pCMV-hAPOBEC3G-XTEN-D10A-SGGS-UGI-SGGS-NLS
物种来源: H. sapiens (human); S. pyogenes and Bacteriophage PBS2
插入大小: 5598 bp
启动子: CMV
克隆方法: 限制性内切酶
5’克隆位点: unknown (unknown if destroyed)
3’克隆位点: NotI (未破坏)
5’测序引物 CMV
3’测序引物 BGH poly reverse

文献引用方法
材料和方法部分: pCMV-hA3G-BE3 was a gift from Jia Chen (Addgene plasmid # 113415 ; http://n2t.net/addgene:113415 ; RRID:Addgene_113415)
参考文献部分: Efficient base editing in methylated regions with a human APOBEC3A-Cas9 fusion. Wang X, Li J, Wang Y, Yang B, Wei J, Wu J, Wang R, Huang X, Chen J, Yang L. Nat Biotechnol. 2018 Aug 20. pii: nbt.4198. doi: 10.1038/nbt.4198. 10.1038/nbt.4198 PubMed 30125268

载体图谱质粒图谱和多克隆位点信息

pCMV-hA3G-BE3载体图谱

载体简介

载体序列

LOCUS       pCMV-hA3G-BE3                8997 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses hA3G-BE3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-hA3G-BE3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8997)
  AUTHORS   Wang X, Li J, Wang Y, Yang V, Wei J, Wu J, Wang R, Huang X, Chen J, 
            Yang L
  TITLE     Efficient base editing in methylated regions with a human 
            APOBEC3A-Cas9 fusion.
  JOURNAL   Nat Biotechnol.,  2018
REFERENCE   2  (bases 1 to 8997)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8997
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(28..47)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        219..598
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        599..802
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     753..773
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     844..863
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        844..862
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2076..6176
                     /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             6189..6437
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6450..6470
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6479..6496
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6519..6536)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6525..6749
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6820..6836)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6820..6836)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6833..6855)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6844..6860
                     /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(6868..6898)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6913..6934
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7051..7068)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7222..7810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7302..7321)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7981..8841)
                     /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     8604..8623
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8842..8946)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcccgatccc ctagggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca
       61 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
      121 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
      181 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
      241 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      301 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      361 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      421 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      481 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      541 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      601 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      661 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      721 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      781 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctaga gatccgcggc
      841 cgctaatacg actcactata gggagagccg ccaccatgaa gcctcacttc agaaacacag
      901 tggagcgaat gtatcgagac acattctcct acaactttta taatagaccc atcctttctc
      961 gtcggaatac cgtctggctg tgctacgaag tgaaaacaaa gggtccctca aggccccctt
     1021 tggacgcaaa gatctttcga ggccaggtgt attccgaact taagtaccac ccagagatga
     1081 gattcttcca ctggttcagc aagtggagga agctgcatcg tgaccaggag tatgaggtca
     1141 cctggtacat atcctggagc ccctgcacaa agtgtacaag ggatatggcc acgttcctgg
     1201 ccgaggaccc gaaggttacc ctgaccatct ttgttgcccg cctctactac ttctgggacc
     1261 cagattacca ggaggcgctt cgcagcctgt gtcagaaaag agacggtccg cgtgccacca
     1321 tgaagatcat gaattatgac gaatttcagc actgttggag caagttcgtg tacagccaaa
     1381 gagagctatt tgagccttgg aataatctgc ctaaatatta tatattactg cacatcatgc
     1441 tgggggagat tctcagacac tcgatggatc cacccacatt cactttcaac tttaacaatg
     1501 aaccttgggt cagaggacgg catgagactt acctgtgtta tgaggtggag cgcatgcaca
     1561 atgacacctg ggtcctgctg aaccagcgca ggggctttct atgcaaccag gctccacata
     1621 aacacggttt ccttgaaggc cgccatgcag agctgtgctt cctggacgtg attccctttt
     1681 ggaagctgga cctggaccag gactacaggg ttacctgctt cacctcctgg agcccctgct
     1741 tcagctgtgc ccaggaaatg gctaaattca tttcaaaaaa caaacacgtg agcctgtgca
     1801 tcttcactgc ccgcatctat gatgatcaag gaagatgtca ggaggggctg cgcaccctgg
     1861 ccgaggctgg ggccaaaatt tcaataatga catacagtga atttaagcac tgctgggaca
     1921 cctttgtgga ccaccaggga tgtcccttcc agccctggga tggactagat gagcacagcc
     1981 aagacctgag tgggaggctg cgggccattc tccagaatca ggaaaacagc ggcagcgaga
     2041 ctcccgggac ctcagagtcc gccacacccg aaagtgataa aaagtattct attggtttag
     2101 ccatcggcac taattccgtt ggatgggctg tcataaccga tgaatacaaa gtaccttcaa
     2161 agaaatttaa ggtgttgggg aacacagacc gtcattcgat taaaaagaat cttatcggtg
     2221 ccctcctatt cgatagtggc gaaacggcag aggcgactcg cctgaaacga accgctcgga
     2281 gaaggtatac acgtcgcaag aaccgaatat gttacttaca agaaattttt agcaatgaga
     2341 tggccaaagt tgacgattct ttctttcacc gtttggaaga gtccttcctt gtcgaagagg
     2401 acaagaaaca tgaacggcac cccatctttg gaaacatagt agatgaggtg gcatatcatg
     2461 aaaagtaccc aacgatttat cacctcagaa aaaagctagt tgactcaact gataaagcgg
     2521 acctgaggtt aatctacttg gctcttgccc atatgataaa gttccgtggg cactttctca
     2581 ttgagggtga tctaaatccg gacaactcgg atgtcgacaa actgttcatc cagttagtac
     2641 aaacctataa tcagttgttt gaagagaacc ctataaatgc aagtggcgtg gatgcgaagg
     2701 ctattcttag cgcccgcctc tctaaatccc gacggctaga aaacctgatc gcacaattac
     2761 ccggagagaa gaaaaatggg ttgttcggta accttatagc gctctcacta ggcctgacac
     2821 caaattttaa gtcgaacttc gacttagctg aagatgccaa attgcagctt agtaaggaca
     2881 cgtacgatga cgatctcgac aatctactgg cacaaattgg agatcagtat gcggacttat
     2941 ttttggctgc caaaaacctt agcgatgcaa tcctcctatc tgacatactg agagttaata
     3001 ctgagattac caaggcgccg ttatccgctt caatgatcaa aaggtacgat gaacatcacc
     3061 aagacttgac acttctcaag gccctagtcc gtcagcaact gcctgagaaa tataaggaaa
     3121 tattctttga tcagtcgaaa aacgggtacg caggttatat tgacggcgga gcgagtcaag
     3181 aggaattcta caagtttatc aaacccatat tagagaagat ggatgggacg gaagagttgc
     3241 ttgtaaaact caatcgcgaa gatctactgc gaaagcagcg gactttcgac aacggtagca
     3301 ttccacatca aatccactta ggcgaattgc atgctatact tagaaggcag gaggattttt
     3361 atccgttcct caaagacaat cgtgaaaaga ttgagaaaat cctaaccttt cgcatacctt
     3421 actatgtggg acccctggcc cgagggaact ctcggttcgc atggatgaca agaaagtccg
     3481 aagaaacgat tactccatgg aattttgagg aagttgtcga taaaggtgcg tcagctcaat
     3541 cgttcatcga gaggatgacc aactttgaca agaatttacc gaacgaaaaa gtattgccta
     3601 agcacagttt actttacgag tatttcacag tgtacaatga actcacgaaa gttaagtatg
     3661 tcactgaggg catgcgtaaa cccgcctttc taagcggaga acagaagaaa gcaatagtag
     3721 atctgttatt caagaccaac cgcaaagtga cagttaagca attgaaagag gactacttta
     3781 agaaaattga atgcttcgat tctgtcgaga tctccggggt agaagatcga tttaatgcgt
     3841 cacttggtac gtatcatgac ctcctaaaga taattaaaga taaggacttc ctggataacg
     3901 aagagaatga agatatctta gaagatatag tgttgactct taccctcttt gaagatcggg
     3961 aaatgattga ggaaagacta aaaacatacg ctcacctgtt cgacgataag gttatgaaac
     4021 agttaaagag gcgtcgctat acgggctggg gacgattgtc gcggaaactt atcaacggga
     4081 taagagacaa gcaaagtggt aaaactattc tcgattttct aaagagcgac ggcttcgcca
     4141 ataggaactt tatgcagctg atccatgatg actctttaac cttcaaagag gatatacaaa
     4201 aggcacaggt ttccggacaa ggggactcat tgcacgaaca tattgcgaat cttgctggtt
     4261 cgccagccat caaaaagggc atactccaga cagtcaaagt agtggatgag ctagttaagg
     4321 tcatgggacg tcacaaaccg gaaaacattg taatcgagat ggcacgcgaa aatcaaacga
     4381 ctcagaaggg gcaaaaaaac agtcgagagc ggatgaagag aatagaagag ggtattaaag
     4441 aactgggcag ccagatctta aaggagcatc ctgtggaaaa tacccaattg cagaacgaga
     4501 aactttacct ctattaccta caaaatggaa gggacatgta tgttgatcag gaactggaca
     4561 taaaccgttt atctgattac gacgtcgatc acattgtacc ccaatccttt ttgaaggacg
     4621 attcaatcga caataaagtg cttacacgct cggataagaa ccgagggaaa agtgacaatg
     4681 ttccaagcga ggaagtcgta aagaaaatga agaactattg gcggcagctc ctaaatgcga
     4741 aactgataac gcaaagaaag ttcgataact taactaaagc tgagaggggt ggcttgtctg
     4801 aacttgacaa ggccggattt attaaacgtc agctcgtgga aacccgccaa atcacaaagc
     4861 atgttgcaca gatactagat tcccgaatga atacgaaata cgacgagaac gataagctga
     4921 ttcgggaagt caaagtaatc actttaaagt caaaattggt gtcggacttc agaaaggatt
     4981 ttcaattcta taaagttagg gagataaata actaccacca tgcgcacgac gcttatctta
     5041 atgccgtcgt agggaccgca ctcattaaga aatacccgaa gctagaaagt gagtttgtgt
     5101 atggtgatta caaagtttat gacgtccgta agatgatcgc gaaaagcgaa caggagatag
     5161 gcaaggctac agccaaatac ttcttttatt ctaacattat gaatttcttt aagacggaaa
     5221 tcactctggc aaacggagag atacgcaaac gacctttaat tgaaaccaat ggggagacag
     5281 gtgaaatcgt atgggataag ggccgggact tcgcgacggt gagaaaagtt ttgtccatgc
     5341 cccaagtcaa catagtaaag aaaactgagg tgcagaccgg agggttttca aaggaatcga
     5401 ttcttccaaa aaggaatagt gataagctca tcgctcgtaa aaaggactgg gacccgaaaa
     5461 agtacggtgg cttcgatagc cctacagttg cctattctgt cctagtagtg gcaaaagttg
     5521 agaagggaaa atccaagaaa ctgaagtcag tcaaagaatt attggggata acgattatgg
     5581 agcgctcgtc ttttgaaaag aaccccatcg acttccttga ggcgaaaggt tacaaggaag
     5641 taaaaaagga tctcataatt aaactaccaa agtatagtct gtttgagtta gaaaatggcc
     5701 gaaaacggat gttggctagc gccggagagc ttcaaaaggg gaacgaactc gcactaccgt
     5761 ctaaatacgt gaatttcctg tatttagcgt cccattacga gaagttgaaa ggttcacctg
     5821 aagataacga acagaagcaa ctttttgttg agcagcacaa acattatctc gacgaaatca
     5881 tagagcaaat ttcggaattc agtaagagag tcatcctagc tgatgccaat ctggacaaag
     5941 tattaagcgc atacaacaag cacagggata aacccatacg tgagcaggcg gaaaatatta
     6001 tccatttgtt tactcttacc aacctcggcg ctccagccgc attcaagtat tttgacacaa
     6061 cgatagatcg caaacgatac acttctacca aggaggtgct agacgcgaca ctgattcacc
     6121 aatccatcac gggattatat gaaactcgga tagatttgtc acagcttggg ggtgactctg
     6181 gtggttctac taatctgtca gatattattg aaaaggagac cggtaagcaa ctggttatcc
     6241 aggaatccat cctcatgctc ccagaggagg tggaagaagt cattgggaac aagccggaaa
     6301 gcgatatact cgtgcacacc gcctacgacg agagcaccga cgagaatgtc atgcttctga
     6361 ctagcgacgc ccctgaatac aagccttggg ctctggtcat acaggatagc aacggtgaga
     6421 acaagattaa gatgctctct ggtggttctc ccaagaagaa gaggaaagtc taaccggtca
     6481 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     6541 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     6601 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     6661 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     6721 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     6781 gataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6841 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6901 ctagggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6961 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7021 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     7081 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7141 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7201 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7261 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7321 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7381 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7441 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7501 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7561 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7621 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     7681 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7741 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7801 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7861 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7921 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7981 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     8041 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     8101 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     8161 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8221 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8281 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8341 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8401 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8461 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8521 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8581 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8641 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8701 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8761 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8821 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8881 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8941 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatc
//

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