xCas9(3.7)-BE3

价格:1200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: xCas9(3.7)-BE3 
出品公司: 爱迪基因
目录编号: 108380
存储实验室: David Liu

载体骨架基本信息
载体名称: pCMV
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: 高拷贝
生长条件: 37 ℃

插入基因信息
插入基因名称: xCas9(3.7)-BE3,rAPOBEC1-XTEN-nxCas9(3.7)-UGI-NLS
物种来源: Synthetic
插入大小: 5133 bp
基因突变: A262T, R324L, S409I, E480K, E543D, M694I, E1219V
GenBank编号 --
启动子 --
标签/融合蛋白: NLS(插入件上的N端子)
克隆方法: Gibson克隆
5' 克隆位点: --
3' 克隆位点: --
5' 测序引物 CMV Forward
3' 测序引物 M13

文献引用方法
材料和方法部分: xCas9(3.7)-BE3 was a gift from David Liu (Addgene plasmid # 108380 ; http://n2t.net/addgene:108380 ; RRID:Addgene_108380)
参考文献部分: Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Hu JH, Miller SM, Geurts MH, Tang W, Chen L, Sun N, Zeina CM, Gao X, Rees HA, Lin Z, Liu DR. Nature. 2018 Feb 28. pii: nature26155. doi: 10.1038/nature26155. 10.1038/nature26155 PubMed 29512652

载体图谱质粒图谱和多克隆位点信息

xCas9(3.7)-BE3质粒图谱

载体简介

载体序列

LOCUS        xCas9(3.7)-BE3              8532 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Mammalian expression vector for xCas9(3.7)-BE3.
ACCESSION   .
VERSION     .
KEYWORDS    xCas9(3.7)-BE3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8532)
  AUTHORS   Hu JH, Miller SM, Geurts MH, Tang W, Chen L, Sun N, Zeina CM, Gao X,
            Rees HA, Lin Z, Liu DR
  TITLE     Evolved Cas9 variants with broad PAM compatibility and high DNA 
            specificity.
  JOURNAL   Nature. 2018 Feb 28. pii: nature26155. doi: 10.1038/nature26155.
  PUBMED    29512652
REFERENCE   2  (bases 1 to 8532)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8532
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        74..453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        454..657
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     608..628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     699..718
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        699..717
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             731..1417
                     /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="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             1466..5566
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5579..5827
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5840..5860
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5869..5886
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5909..5926)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5915..6139
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6210..6226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6210..6226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6223..6245)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6234..6250
                     /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(6258..6288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6303..6324
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6441..6458)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6612..7200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6692..6711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7371..8231)
                     /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     7994..8013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8232..8336)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8415..8434)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
       61 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
      661 ttagtgaacc gtcagatccg ctagagatcc gcggccgcta atacgactca ctatagggag
      721 agccgccacc atgagctcag agactggccc agtggctgtg gaccccacat tgagacggcg
      781 gatcgagccc catgagtttg aggtattctt cgatccgaga gagctccgca aggagacctg
      841 cctgctttac gaaattaatt gggggggccg gcactccatt tggcgacata catcacagaa
      901 cactaacaag cacgtcgaag tcaacttcat cgagaagttc acgacagaaa gatatttctg
      961 tccgaacaca aggtgcagca ttacctggtt tctcagctgg agcccatgcg gcgaatgtag
     1021 tagggccatc actgaattcc tgtcaaggta tccccacgtc actctgttta tttacatcgc
     1081 aaggctgtac caccacgctg acccccgcaa tcgacaaggc ctgcgggatt tgatctcttc
     1141 aggtgtgact atccaaatta tgactgagca ggagtcagga tactgctgga gaaactttgt
     1201 gaattatagc ccgagtaatg aagcccactg gcctaggtat ccccatctgt gggtacgact
     1261 gtacgttctt gaactgtact gcatcatact gggcctgcct ccttgtctca acattctgag
     1321 aaggaagcag ccacagctga cattctttac catcgctctt cagtcttgtc attaccagcg
     1381 actgccccca cacattctct gggccaccgg gttgaaaagc ggcagcgaga ctcccgggac
     1441 ctcagagtcc gccacacccg aaagtgacaa gaagtactcc attgggctcg ctatcggcac
     1501 aaacagcgtc ggctgggccg tcattacgga cgagtacaag gtgccgagca aaaaattcaa
     1561 agttctgggc aataccgatc gccacagcat aaagaagaac ctcattggcg ccctcctgtt
     1621 cgactccggg gagacggccg aagccacgcg gctcaaaaga acagcacggc gcagatatac
     1681 ccgcagaaag aatcggatct gctacctgca ggagatcttt agtaatgaga tggctaaggt
     1741 ggatgactct ttcttccata ggctggagga gtcctttttg gtggaggagg ataaaaagca
     1801 cgagcgccac ccaatctttg gcaatatcgt ggacgaggtg gcgtaccatg aaaagtaccc
     1861 aaccatatat catctgagga agaagcttgt agacagtact gataaggctg acttgcggtt
     1921 gatctatctc gcgctggcgc atatgatcaa atttcgggga cacttcctca tcgaggggga
     1981 cctgaaccca gacaacagcg atgtcgacaa actctttatc caactggttc agacttacaa
     2041 tcagcttttc gaagagaacc cgatcaacgc atccggagtt gacgccaaag caatcctgag
     2101 cgctaggctg tccaaatccc ggcggctcga aaacctcatc gcacagctcc ctggggagaa
     2161 gaagaacggc ctgtttggta atcttatcgc cctgtccctc gggctgaccc ccaactttaa
     2221 atctaacttc gacctggccg aagataccaa gcttcaactg agcaaagaca cctacgatga
     2281 tgatctcgac aatctgctgg cccagatcgg cgaccagtac gcagaccttt ttttggcggc
     2341 aaagaacctg tcagacgcca ttctgctgag tgatattctg cgagtgaaca cggagatcac
     2401 caaagctccg ctgagcgcta gtatgatcaa gctctatgat gagcaccacc aagacttgac
     2461 tttgctgaag gcccttgtca gacagcaact gcctgagaag tacaaggaaa ttttcttcga
     2521 tcagtctaaa aatggctacg ccggatacat tgacggcgga gcaagccagg aggaatttta
     2581 caaatttatt aagcccatct tggaaaaaat ggacggcacc gaggagctgc tggtaaagct
     2641 taacagagaa gatctgttgc gcaaacagcg cactttcgac aatggaatca tcccccacca
     2701 gattcacctg ggcgaactgc acgctatcct caggcggcaa gaggatttct accccttttt
     2761 gaaagataac agggaaaaga ttgagaaaat cctcacattt cggataccct actatgtagg
     2821 ccccctcgcc cggggaaatt ccagattcgc gtggatgact cgcaaatcag aagagaccat
     2881 cactccctgg aacttcgaga aagtcgtgga taagggggcc tctgcccagt ccttcatcga
     2941 aaggatgact aactttgata aaaatctgcc taacgaaaag gtgcttccta aacactctct
     3001 gctgtacgag tacttcacag tttataacga gctcaccaag gtcaaatacg tcacagaagg
     3061 gatgagaaag ccagcattcc tgtctggaga tcagaagaaa gctattgtgg acctcctctt
     3121 caagacgaac cggaaagtta ccgtgaaaca gctcaaagaa gactatttca aaaagattga
     3181 atgtttcgac tctgttgaaa tcagcggagt ggaggatcgc ttcaacgcat ccctgggaac
     3241 gtatcacgat ctcctgaaaa tcattaaaga caaggacttc ctggacaatg aggagaacga
     3301 ggacattctt gaggacattg tcctcaccct tacgttgttt gaagataggg agatgattga
     3361 agaacgcttg aaaacttacg ctcatctctt cgacgacaaa gtcatgaagc agctcaagag
     3421 gcgccgatat acaggatggg ggcggctgtc aagaaaactg atcaatggga tccgagacaa
     3481 gcagagtgga aagacaatcc tggattttct taagtccgat ggatttgcca accggaactt
     3541 cattcagttg atccatgatg actctctcac ctttaaggag gacatccaga aagcacaagt
     3601 ttctggccag ggggacagtc ttcacgagca catcgctaat cttgcaggta gcccagctat
     3661 caaaaaggga atactgcaga ccgttaaggt cgtggatgaa ctcgtcaaag taatgggaag
     3721 gcataagccc gagaatatcg ttatcgagat ggcccgagag aaccaaacca cccagaaggg
     3781 acagaagaac agtagggaaa ggatgaagag gattgaagag ggtataaaag aactggggtc
     3841 ccaaatcctt aaggaacacc cagttgaaaa cacccagctt cagaatgaga agctctacct
     3901 gtactacctg cagaacggca gggacatgta cgtggatcag gaactggaca tcaatcggct
     3961 ctccgactac gacgtggatc atatcgtgcc ccagtctttt ctcaaagatg attctattga
     4021 taataaagtg ttgacaagat ccgataaaaa cagagggaag agtgataacg tcccctcaga
     4081 agaagttgtc aagaaaatga aaaattattg gcggcagctg ctgaacgcca aactgatcac
     4141 acaacggaag ttcgataatc tgactaaggc tgaacgaggt ggcctgtctg agttggataa
     4201 agccggtttc atcaaaaggc agcttgttga gacacgccag atcaccaagc acgtggccca
     4261 aattctcgat tcacgcatga acaccaagta cgatgaaaat gacaaactga ttcgagaggt
     4321 gaaagttatt actctgaagt ctaagctggt ctcagatttc agaaaggact ttcagtttta
     4381 taaggtgaga gagatcaaca attaccacca tgcgcatgat gcctacctga atgcagtggt
     4441 aggcactgca cttatcaaaa aatatcccaa gcttgaatct gaatttgttt acggagacta
     4501 taaagtgtac gatgttagga aaatgatcgc aaagtctgag caggaaatag gcaaggccac
     4561 cgctaagtac ttcttttaca gcaatattat gaattttttc aagaccgaga ttacactggc
     4621 caatggagag attcggaagc gaccacttat cgaaacaaac ggagaaacag gagaaatcgt
     4681 gtgggacaag ggtagggatt tcgcgacagt ccggaaggtc ctgtccatgc cgcaggtgaa
     4741 catcgttaaa aagaccgaag tacagaccgg aggcttctcc aaggaaagta tcctcccgaa
     4801 aaggaacagc gacaagctga tcgcacgcaa aaaagattgg gaccccaaga aatacggcgg
     4861 attcgattct cctacagtcg cttacagtgt actggttgtg gctaaagtgg agaaagggaa
     4921 gtctaaaaaa ctcaaaagcg tcaaggaact gctgggcatc acaatcatgg agcgatcaag
     4981 cttcgaaaaa aaccccatcg actttctcga ggcgaaagga tataaagagg tcaaaaaaga
     5041 cctcatcatt aagcttccca agtactctct ctttgagctt gaaaacggcc ggaaacgaat
     5101 gctcgctagt gcgggcgtgc tgcagaaagg taacgagctg gcactgccct ctaaatacgt
     5161 taatttcttg tatctggcca gccactatga aaagctcaaa gggtctcccg aagataatga
     5221 gcagaagcag ctgttcgtgg aacaacacaa acactacctt gatgagatca tcgagcaaat
     5281 aagcgaattc tccaaaagag tgatcctcgc cgacgctaac ctcgataagg tgctttctgc
     5341 ttacaataag cacagggata agcccatcag ggagcaggca gaaaacatta tccacttgtt
     5401 tactctgacc aacttgggcg cgcctgcagc cttcaagtac ttcgacacta ccatagacag
     5461 aaagcggtac acctctacaa aggaggtcct ggacgccaca ctgattcatc agtcaattac
     5521 ggggctctat gaaacaagaa tcgacctctc tcagctcggt ggagactctg gtggttctac
     5581 taatctgtca gatattattg aaaaggagac cggtaagcaa ctggttatcc aggaatccat
     5641 cctcatgctc ccagaggagg tggaagaagt cattgggaac aagccggaaa gcgatatact
     5701 cgtgcacacc gcctacgacg agagcaccga cgagaatgtc atgcttctga ctagcgacgc
     5761 ccctgaatac aagccttggg ctctggtcat acaggatagc aacggtgaga acaagattaa
     5821 gatgctctct ggtggttctc ccaagaagaa gaggaaagtc taaccggtca tcatcaccat
     5881 caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     5941 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     6001 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     6061 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     6121 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc gataccgtcg
     6181 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6241 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctagggtgcc
     6301 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6361 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6421 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6481 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6541 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6601 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6661 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6721 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6781 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6841 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6901 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6961 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7021 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     7081 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7141 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7201 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7261 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7321 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7381 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7441 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7501 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7561 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7621 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7681 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7741 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7801 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7861 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7921 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7981 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8041 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8101 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8161 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8221 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8281 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8341 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatctcc cgatccccta
     8401 gggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     8461 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     8521 ggcttgaccg ac
//

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