pCMV-MMLVgag-3xNES-ABE8e

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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: pCMV-MMLVgag-3xNES-ABE8e
目录编号: 181751
存储实验室: David Liu

载体骨架基本信息
载体名称: pCDNA3-like
空载体大小: 4757 bp
完整质粒大小: 11365bp
载体类型: 哺乳细胞表达
载体抗性: Ampicillin, 100 μg/mL
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: MMLVgag-3xNES-ABE8e
物种来源: H. sapiens (human), M. musculus (mouse)
插入大小: 6609 bp
启动子: CMV
标签/融合蛋白: Flag (N terminal on insert)
克隆方法: Ligation Independent Cloning
5’测序引物 ACCCTGGTCATCATCCTG
3’测序引物 CAGGAAACAGCTATGAC; CCTTCCGAGTGAGAGAC

文献引用方法
材料和方法部分: pCMV-MMLVgag-3xNES-ABE8e was a gift from David Liu (Addgene plasmid # 181751 ; http://n2t.net/addgene:181751 ; RRID:Addgene_181751)
参考文献部分: Engineered virus-like particles for efficient in vivo delivery of therapeutic proteins. Banskota S, Raguram A, Suh S, Du SW, Davis JR, Choi EH, Wang X, Nielsen SC, Newby GA, Randolph PB, Osborn MJ, Musunuru K, Palczewski K, Liu DR. Cell. 2022 Jan 7. pii: S0092-8674(21)01484-7. doi: 10.1016/j.cell.2021.12.021. 10.1016/j.cell.2021.12.021 PubMed 35

载体图谱质粒图谱和多克隆位点信息

pDY0216

载体简介

载体序列

LOCUS       pCMV-MMLVgag-3xNES-ABE8e               11365 bp ds-DNA     circular SYN 08-FEB-2022
DEFINITION  MMLV-Gag fused to 3xNES-ABE8e with TSTLLMENSS cleavage site between 
            Gag-NES and ABE. For production of virus like particles with 
            base-editor RNP cargo..
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-MMLVgag-3xNES-ABE8e
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11365)
  AUTHORS   Banskota S, Raguram A, Suh S, Du SW, Davis JR, Choi EH, Wang X, 
            Nielsen SC, Newby GA, Randolph PB, Osborn MJ, Musunuru K, Palczewski
            K, Liu DR
  TITLE     Engineered virus-like particles for efficient in vivo delivery of 
            therapeutic proteins.
  JOURNAL   Cell. 2022 Jan 7. pii: S0092-8674(21)01484-7. doi: 
            10.1016/j.cell.2021.12.021.
  PUBMED    35021064
REFERENCE   2  (bases 1 to 11365)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 8, 2022 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..11365
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             4..420
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     350..372
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     388..404
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1633..1659
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1672..1698
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1711..1737
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1771..1794
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1828..1848
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2443..6543
                     /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             6586..6606
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6612..6631)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6677..6732
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6731..6750)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7090..7106)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7090..7106)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7103..7125)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7114..7130
                     /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(7138..7168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7183..7204
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7321..7338)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7492..8080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7572..7591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8251..9111)
                     /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     8874..8893
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9112..9216)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9242..9697)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9379..9400)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9591..9610
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9824..9846
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9838..9855
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9839..9855
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9865..9884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9865..9883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        10004..10383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10384..10587
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10538..10558
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     10584..10608
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          10699..11271
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
ORIGIN
        1 atgggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag
       61 cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct
      121 gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc
      181 atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc
      241 ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt
      301 gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct
      361 cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc
      421 aaacctaaac ctcaagttct ttctgacagt ggggggccgc tcatcgacct acttacagaa
      481 gaccccccgc cttataggga cccaagacca cccccttccg acagggacgg aaatggtgga
      541 gaagcgaccc ctgcgggaga ggcaccggac ccctccccaa tggcatctcg cctacgtggg
      601 agacgggagc cccctgtggc cgactccact acctcgcagg cattccccct ccgcgcagga
      661 ggaaacggac agcttcaata ctggccgttc tcctcttctg acctttacaa ctggaaaaat
      721 aataaccctt ctttttctga agatccaggt aaactgacag ctctgatcga gtctgtcctc
      781 atcacccatc agcccacctg ggacgactgt cagcagctgt tggggactct gctgaccgga
      841 gaagaaaaac aacgggtgct cttagaggct agaaaggcgg tgcggggcga tgatgggcgc
      901 cccactcaac tgcccaatga agtcgatgcc gcttttcccc tcgagcgccc agactgggat
      961 tacaccaccc aggcaggtag gaaccaccta gtccactatc gccagttgct cctagcgggt
     1021 ctccaaaacg cgggcagaag ccccaccaat ttggccaagg taaaaggaat aacacaaggg
     1081 cccaatgagt ctccctcggc cttcctagag agacttaagg aagcctatcg caggtacact
     1141 ccttatgacc ctgaggaccc agggcaagaa actaatgtgt ctatgtcttt catttggcag
     1201 tctgccccag acattgggag aaagttagag aggttagaag atttaaaaaa caagacgctt
     1261 ggagatttgg ttagagaggc agaaaagatc tttaataaac gagaaacccc ggaagaaaga
     1321 gaggaacgta tcaggagaga aacagaggaa aaagaagaac gccgtaggac agaggatgag
     1381 cagaaagaga aagaaagaga tcgtaggaga catagagaga tgagcaagct attggccact
     1441 gtcgttagtg gacagaaaca ggatagacag ggaggagaac gaaggaggtc ccaactcgat
     1501 cgcgaccagt gtgcctactg caaagaaaag gggcactggg ctaaagattg tcccaagaaa
     1561 ccacgaggac ctcggggacc aagaccccag acctccctcc tgaccctaga tgactctggc
     1621 ggctcacttc aactgcctcc acttgaaaga ctgacactgg gatcattaca attacctcct
     1681 ttagaacgat taacactcgg ttcactacag cttccgcctc ttgagagatt gacattaacg
     1741 tccacgctgc taatggagaa ctcgtctgga gattacaaag acgatgacga taagaaacgg
     1801 acagccgacg gaagcgagtt cgagtcacca aagaagaagc ggaaagtctc tgaggtggag
     1861 ttttcccacg agtactggat gagacatgcc ctgaccctgg ccaagagggc acgggatgag
     1921 agggaggtgc ctgtgggagc cgtgctggtg ctgaacaata gagtgatcgg cgagggctgg
     1981 aacagagcca tcggcctgca cgacccaaca gcccatgccg aaattatggc cctgagacag
     2041 ggcggcctgg tcatgcagaa ctacagactg attgacgcca ccctgtacgt gacattcgag
     2101 ccttgcgtga tgtgcgccgg cgccatgatc cactctagga tcggccgcgt ggtgtttggc
     2161 gtgaggaact caaaaagagg cgccgcaggc tccctgatga acgtgctgaa ctaccccggc
     2221 atgaatcacc gcgtcgaaat taccgaggga atcctggcag atgaatgtgc cgccctgctg
     2281 tgcgatttct atcggatgcc tagacaggtg ttcaatgctc agaagaaggc ccagagctcc
     2341 atcaactccg gaggatctag cggaggctcc tctggctctg agacacctgg cacaagcgag
     2401 agcgcaacac ctgaaagcag cgggggcagc agcggggggt cagacaagaa gtacagcatc
     2461 ggcctggcca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg
     2521 cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg
     2581 atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc
     2641 gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc
     2701 aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg
     2761 gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc
     2821 taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac
     2881 aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac
     2941 ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag
     3001 ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac
     3061 gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc
     3121 cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc
     3181 ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc
     3241 aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc
     3301 gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga
     3361 gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag
     3421 caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac
     3481 aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc
     3541 agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag
     3601 gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac
     3661 ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa
     3721 gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc
     3781 atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga
     3841 aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc
     3901 gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg
     3961 ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg
     4021 aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc
     4081 atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac
     4141 tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc
     4201 aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg
     4261 gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag
     4321 gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg
     4381 atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc
     4441 aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc
     4501 ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac
     4561 atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg
     4621 gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc
     4681 gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac
     4741 cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc
     4801 atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag
     4861 aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa
     4921 ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg
     4981 aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc
     5041 gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg
     5101 aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc
     5161 ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc
     5221 acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac
     5281 aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg
     5341 aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc
     5401 tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag
     5461 tttgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag
     5521 gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag
     5581 accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc
     5641 gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg
     5701 agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa
     5761 gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac
     5821 cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc
     5881 aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc
     5941 atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac
     6001 aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa
     6061 aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc
     6121 ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc
     6181 tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac
     6241 gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg
     6301 gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag
     6361 aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt
     6421 gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg
     6481 atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggt
     6541 gactctggcg gctcaaaaag aaccgccgac ggcagcgaat tcgagcccaa gaagaagagg
     6601 aaagtctgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     6661 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     6721 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     6781 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg
     6841 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     6901 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     6961 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     7021 ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacggatc ggccgcaatt
     7081 cgtaaccatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     7141 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     7201 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     7261 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     7321 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     7381 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7441 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7501 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7561 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7621 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7681 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7741 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7801 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7861 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7921 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7981 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     8041 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     8101 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8161 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8221 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8281 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8341 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8401 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8461 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8521 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8581 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8641 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8701 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8761 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8821 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8881 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8941 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     9001 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     9061 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     9121 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9181 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     9241 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     9301 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     9361 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     9421 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     9481 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     9541 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     9601 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     9661 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     9721 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     9781 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     9841 aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt ggagctccac
     9901 cgcggtggcg gccgctctag agagcttggc ccattgcata cgttgtatcc atatcataat
     9961 atgtacattt atattggctc atgtccaaca ttaccgccat gttgacattg attattgact
    10021 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
    10081 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
    10141 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
    10201 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
    10261 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
    10321 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
    10381 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
    10441 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
    10501 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
    10561 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc
    10621 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccggtcgacc
    10681 gatcctgaga acttcagggt gagtttgggg acccttgatt gttctttctt tttcgctatt
    10741 gtaaaattca tgttatatgg agggggcaaa gttttcaggg tgttgtttag aatgggaaga
    10801 tgtcccttgt atcaccatgg accctcatga taattttgtt tctttcactt tctactctgt
    10861 tgacaaccat tgtctcctct tattttcttt tcattttctg taactttttc gttaaacttt
    10921 agcttgcatt tgtaacgaat ttttaaattc acttttgttt atttgtcaga ttgtaagtac
    10981 tttctctaat cacttttttt tcaaggcaat cagggtatat tatattgtac ttcagcacag
    11041 ttttagagaa caattgttat aattaaatga taaggtagaa tatttctgca tataaattct
    11101 ggctggcgtg gaaatattct tattggtaga aacaactaca ccctggtcat catcctgcct
    11161 ttctctttat ggttacaatg atatacactg tttgagatga ggataaaata ctctgagtcc
    11221 aaaccgggcc cctctgctaa ccatgttcat gccttcttct ctttcctaca gctcctgggc
    11281 aacgtgctgg ttgttgtgct gtctcatcat tttggcaaag aattcctcga gggccggatc
    11341 cgaattcctg tatttgtctg aaaac
//

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