CP1028-CBEmax

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CP1028CBEmax载体基本信息

质粒名称: CP1028-CBEmax 
出品公司: 爱迪基因
目录编号: 119802
存储实验室: David Liu

载体骨架基本信息
载体名称: pCMV with a BR322 origin
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: 哺乳细胞表达
筛选标记: --
高拷贝/低拷贝: --
生长条件: 37 ℃

插入基因信息
插入基因名称: rApobec1-nSpCas9 (CP1028) -UGI-UGI
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: --
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: CP1028-CBEmax was a gift from David Liu (Addgene plasmid # 119802 ; http://n2t.net/addgene:119802 ; RRID:Addgene_119802)
参考文献部分: Circularly permuted and PAM-modified Cas9 variants broaden the targeting scope of base editors. Huang TP, Zhao KT, Miller SM, Gaudelli NM, Oakes BL, Fellmann C, Savage DF, Liu DR. Nat Biotechnol. 2019 May 20. pii: 10.1038/s41587-019-0134-y. doi: 10.1038/s41587-019-0134-y. 10.1038/s41587-019-0134-y PubMed 31110355

CP1028CBEmax载体图谱质粒图谱和多克隆位点信息

CP1028-CBEmax质粒图谱

CP1028CBEmax载体简介

CP1028CBEmax载体序列

LOCUS       CP1028-CBEmax               9024 bp ds-DNA     circular SYN 19-DEC-2018
DEFINITION  C-to-T base editor.
ACCESSION   .
VERSION     .
KEYWORDS    CP1028-CBEmax
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9024)
  TITLE     Liu lab base editing plasmids
REFERENCE   2  (bases 1 to 9024)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 19, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9024
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        162..541
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        542..745
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     696..716
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     787..806
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        787..805
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             855..875
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             876..1559
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
                     RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
                     RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
                     AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
                     WATGLK"
     CDS             2739..4457
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             5850..6098
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6129..6377
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6420..6440
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6449..6466
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6489..6506)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6495..6719
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6790..6806)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6790..6806)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6803..6825)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6814..6830
                     /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(6838..6868)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6883..6904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7021..7038)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7192..7780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7272..7291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7951..8811)
                     /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     8574..8593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8812..8916)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8995..9014)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
       61 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
      121 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
      181 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      241 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      301 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      361 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      421 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      481 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      541 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      601 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      661 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      721 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agagatccgc
      781 ggccgctaat acgactcact atagggagag ccgccaccat gaaacggaca gccgacggaa
      841 gcgagttcga gtcaccaaag aagaagcgga aagtctcctc agagactggg cctgtcgccg
      901 tcgatccaac cctgcgccgc cggattgaac ctcacgagtt tgaagtgttc tttgaccccc
      961 gggagctgag aaaggagaca tgcctgctgt acgagatcaa ctggggaggc aggcactcca
     1021 tctggaggca cacctctcag aacacaaata agcacgtgga ggtgaacttc atcgagaagt
     1081 ttaccacaga gcggtacttc tgccccaata ccagatgtag catcacatgg tttctgagct
     1141 ggtccccttg cggagagtgt agcagggcca tcaccgagtt cctgtccaga tatccacacg
     1201 tgacactgtt tatctacatc gccaggctgt atcaccacgc agacccaagg aataggcagg
     1261 gcctgcgcga tctgatcagc tccggcgtga ccatccagat catgacagag caggagtccg
     1321 gctactgctg gcggaacttc gtgaattatt ctcctagcaa cgaggcccac tggcctaggt
     1381 acccacacct gtgggtgcgc ctgtacgtgc tggagctgta ttgcatcatc ctgggcctgc
     1441 ccccttgtct gaatatcctg cggagaaagc agccccagct gaccttcttt acaatcgccc
     1501 tgcagtcttg tcactatcag aggctgccac cccacatcct gtgggccaca ggcctgaagt
     1561 ctggaggatc tagcggagga tcctctggca gcgagacacc aggaacaagc gagtcagcaa
     1621 caccagagag cagtggcggc agcagcggcg gcagcgaaat cggcaaggct accgccaagt
     1681 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
     1741 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
     1801 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
     1861 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
     1921 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
     1981 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
     2041 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
     2101 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
     2161 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
     2221 ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
     2281 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
     2341 agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt
     2401 tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca
     2461 agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga
     2521 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt
     2581 acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
     2641 acgagacacg gatcgacctg tctcagctgg gaggcgacgg cggatcagga ggctctggcg
     2701 gttcaggtgg atcaggcggt agcggaggtt caggtggtat ggacaagaag tacagcatcg
     2761 gcctggccat cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc
     2821 ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga
     2881 tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg
     2941 ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca
     3001 acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg
     3061 aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct
     3121 accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca
     3181 aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact
     3241 tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc
     3301 tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg
     3361 ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc
     3421 agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc
     3481 tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca
     3541 aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg
     3601 acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag
     3661 tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc
     3721 accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca
     3781 aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca
     3841 gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg
     3901 aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg
     3961 gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag
     4021 atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca
     4081 tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa
     4141 agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg
     4201 cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc
     4261 tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga
     4321 aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca
     4381 tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact
     4441 acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca
     4501 acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg
     4561 acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg
     4621 acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga
     4681 tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca
     4741 acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct
     4801 tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca
     4861 tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg
     4921 ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg
     4981 tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc
     5041 agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca
     5101 tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga
     5161 acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac
     5221 tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag agctttctga
     5281 aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg
     5341 acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga
     5401 acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc
     5461 tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca
     5521 caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca
     5581 agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga
     5641 aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct
     5701 acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt
     5761 tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcaga
     5821 gcggcgggag cggcgggagc ggggggagca ctaatctgag cgacatcatt gagaaggaga
     5881 ctgggaaaca gctggtcatt caggagtcca tcctgatgct gcctgaggag gtggaggaag
     5941 tgatcggcaa caagccagag tctgacatcc tggtgcacac cgcctacgac gagtccacag
     6001 atgaaaatgt gatgctgctg acctctgacg cccccgagta taagccttgg gccctggtca
     6061 tccaggattc taacggcgag aataagatca agatgctgag cggaggatcc ggaggatctg
     6121 gaggcagcac caacctgtct gacatcatcg agaaggagac aggcaagcag ctggtcatcc
     6181 aggagagcat cctgatgctg cccgaagaag tcgaagaagt gatcggaaac aagcctgaga
     6241 gcgatatcct ggtccatacc gcctacgacg agagtaccga cgaaaatgtg atgctgctga
     6301 catccgacgc cccagagtat aagccctggg ctctggtcat ccaggattcc aacggagaga
     6361 acaaaatcaa aatgctgtct ggcggctcaa aaagaaccgc cgacggcagc gaattcgagc
     6421 ccaagaagaa gaggaaagtc taaccggtca tcatcaccat caccattgag tttaaacccg
     6481 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     6541 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     6601 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     6661 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     6721 ttctgaggcg gaaagaacca gctggggctc gataccgtcg acctctagct agagcttggc
     6781 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6841 catacgagcc ggaagcataa agtgtaaagc ctagggtgcc taatgagtga gctaactcac
     6901 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6961 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     7021 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     7081 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7141 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7201 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7261 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7321 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7381 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7441 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7501 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7561 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7621 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7681 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7741 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7801 tacggggtct gacactcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7861 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7921 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7981 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8041 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8101 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8161 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8221 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8281 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8341 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8401 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8461 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8521 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8581 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8641 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8701 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8761 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8821 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8881 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8941 tgacgtcgac ggatcgggag atcgatctcc cgatccccta gggtcgactc tcagtacaat
     9001 ctgctctgat gccgcatagt taag
//

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