pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro

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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: pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro
出品公司: Addgene
目录编号: 141385
存储实验室: Nevan Krogan

载体骨架基本信息
载体名称: pLVX-EF1alpha-IRES-Puro
载体出品公司: Takara Bio
空载体大小: 8825 bp
完整质粒大小: 9138 bp
载体类型: 哺乳细胞表达, Lentiviral
筛选标记: Puromycin
载体抗性: 氨苄霉素
生长温度: 30 ℃
宿主菌: NEB Stable
拷贝数: 未知

插入基因信息
插入基因名称: SARS-CoV-2-E
物种来源: SARS-CoV-2
基因突变: human codon optimized
启动子: EF1alpha
标签/融合蛋白: 2xStrep (C terminal on insert)
克隆方法: 未知

文献引用方法
材料和方法部分:
pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro was a gift from Nevan Krogan (Addgene plasmid # 141385 ; http://n2t.net/addgene:141385 ; RRID:Addgene_141385)
For viral preps, please replace (Addgene plasmid # 141385) in the above sentence with: (Addgene viral prep # 141385-LV)
参考文献部分: A SARS-CoV-2 protein interaction map reveals targets for drug repurposing. Gordon DE, Jang GM, Bouhaddou M, Xu J, Obernier K, White KM, O'Meara MJ, Rezelj VV, Guo JZ, Swaney DL, Tummino TA, Huttenhain R, Kaake RM, Richards AL, Tutuncuoglu B, Foussard H, Batra J, Haas K, Modak M, Kim M, Haas P, Polacco BJ, Braberg H, Fabius JM, Eckhardt M, Soucheray M, Bennett MJ, Cakir M, McGregor MJ, Li Q, Meyer B, Roesch F, Vallet T, Mac Kain A, Miorin L, Moreno E, Naing ZZC, Zhou Y, Peng S, Shi Y, Zhang Z, Shen W, Kirby IT, Melnyk JE, Chorba JS, Lou K, Dai SA, Barrio-Hernandez I, Memon D, Hernandez-Armenta C, Lyu J, Mathy CJP, Perica T, Pilla KB, Ganesan SJ, Saltzberg DJ, Rakesh R, Liu X, Rosenthal SB, Calviello L, Venkataramanan S, Liboy-Lugo J, Lin Y, Huang XP, Liu Y, Wankowicz SA, Bohn M, Safari M, Ugur FS, Koh C, Savar NS, Tran QD, Shengjuler D, Fletcher SJ, O'Neal MC, Cai Y, Chang JCJ, Broadhurst DJ, Klippsten S, Sharp PP, Wenzell NA, Kuzuoglu-Ozturk D, Wang HY, Trenker R, Young JM, Cavero DA, Hiatt J, Roth TL, Rathore U, Subramanian A, Noack J, Hubert M, Stroud RM, Frankel AD, Rosenberg OS, Verba KA, Agard DA, Ott M, Emerman M, Jura N, von Zastrow M, Verdin E, Ashworth A, Schwartz O, d'Enfert C, Mukherjee S, Jacobson M, Malik HS, Fujimori DG, Ideker T, Craik CS, Floor SN, Fraser JS, Gross JD, Sali A, Roth BL, Ruggero D, Taunton J, Kortemme T, Beltrao P, Vignuzzi M, Garcia-Sastre A, Shokat KM, Shoichet BK, Krogan NJ. Nature. 2020 Jul;583(7816):459-468. doi: 10.1038/s41586-020-2286-9. Epub 2020 Apr 30. 10.1038/s41586-020-2286-9 PubMed 32353859

载体图谱质粒图谱和多克隆位点信息

pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro载体图谱

载体简介

载体序列

LOCUS      pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro                9138 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Lentiviral expression of SARS-CoV-2 envelope (E) protein; transient 
            expression and generate lentivirus.
ACCESSION   .
VERSION     .
KEYWORDS    pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9138)
  AUTHORS   Gordon DE, et al.
  TITLE     A SARS-CoV-2-Human Protein-Protein Interaction Map Reveals Drug 
            Targets and Potential Drug Repurposing
  JOURNAL   BioRxiv March 2020
REFERENCE   2  (bases 1 to 9138)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9138
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..1182
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          231..1173
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1130..1150
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1456..1479
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1516..1539
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    1551..2124
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1718..1735)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1945..1964
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2157..2756
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2157..2176)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2653..2673
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2770..3358
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2823..2843)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3241..3252)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3565..4198
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4326..4342)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4326..4342)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4339..4361)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4350..4366
                     /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(4374..4404)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4419..4440
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4557..4574)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4728..5316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4808..4827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5487..6347)
                     /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     6110..6129
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6348..6452)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    6500..6634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6537..6556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6591..6610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             6802..7435
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7482..7607
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8104..8337
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8522..8566
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    8829..8944
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
ORIGIN
        1 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
       61 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
      121 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
      181 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
      241 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
      301 cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
      361 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
      421 tggcttgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
      481 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
      541 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
      601 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
      661 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
      721 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
      781 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
      841 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
      901 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
      961 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     1021 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     1081 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     1141 acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggatctat ttccggtgaa
     1201 ttcgccgcca ccatgtacag cttcgtatca gaagaaaccg ggacactgat cgtaaattct
     1261 gtgctcttgt ttctggcatt cgtcgtattt ctcctcgtca cactggcaat tctgactgca
     1321 ttgaggcttt gcgcctactg ttgtaacatt gtcaatgtat ctctcgtgaa accctcattc
     1381 tacgtttaca gcagggtgaa gaatctcaat tctagcaggg tgccggatct cctcgttctc
     1441 gaaggcggcg ggggatggag ccatccacaa ttcgagaaag gcggtggttc aggaggaggt
     1501 agcgggggtg gatcatggtc acatccgcag tttgaaaagt aaggatcccg cccctctccc
     1561 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     1621 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     1681 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     1741 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     1801 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     1861 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     1921 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     1981 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     2041 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     2101 ttcctttgaa aaacacgatg ataagcttgc cacaacccac aaggagacga ccttccatga
     2161 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
     2221 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
     2281 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     2341 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     2401 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     2461 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     2521 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     2581 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     2641 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     2701 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctagacgc
     2761 gtctggaaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2821 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2881 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2941 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3001 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3061 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3121 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     3181 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3241 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3301 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     3361 ggaattaatt ctgcagtcga gacctagaaa aacatggagc aatcacaagt agcaatacag
     3421 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc
     3481 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     3541 actttttaaa agaaaagagg ggactggaag ggctaattca ctcccaacga agacaagata
     3601 tccttgatct gtggatctac cacacacaag gctacttccc tgattagcag aactacacac
     3661 cagggccagg ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg
     3721 agccagataa ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga
     3781 gcctgcatgg gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc
     3841 tagcatttca tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc
     3901 gagcttgcta caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga
     3961 ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg
     4021 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     4081 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     4141 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     4201 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     4261 agtgagaggc cttgacattg ctagcgttta ccgtcgacct ctagctagag cttggcgtaa
     4321 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4381 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4441 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4501 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4561 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4621 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4681 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4741 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4801 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4861 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4921 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4981 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5041 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5101 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5161 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5221 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5281 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5341 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5401 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5461 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5521 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5581 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5641 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5701 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5761 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5821 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5881 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5941 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6001 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6061 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6121 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6181 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6241 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6301 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6361 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6421 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6481 gtcgacggat cgggagatca acttgtttat tgcagcttat aatggttaca aataaagcaa
     6541 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6601 caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa
     6661 tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt
     6721 actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat
     6781 atgtactaca aacttagtag ttggaagggc taattcactc ccaaagaaga caagatatcc
     6841 ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag
     6901 ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc
     6961 cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc
     7021 tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag
     7081 catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag
     7141 cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg
     7201 gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct
     7261 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     7321 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     7381 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     7441 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     7501 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     7561 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     7621 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     7681 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     7741 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     7801 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     7861 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     7921 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     7981 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     8041 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     8101 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     8161 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     8221 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     8281 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     8341 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     8401 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     8461 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     8521 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     8581 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     8641 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     8701 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     8761 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg
     8821 gtatcgcctt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
     8881 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
     8941 attttcgggt ttattacagg gacagcagag atccagttta tcgatgagta attcatacaa
     9001 aaggactcgc ccctgccttg gggaatccca gggaccgtcg ttaaactccc actaacgtag
     9061 aacccagaga tcgctgcgtt cccgccccct cacccgcccg ctctcgtcat cactgaggtg
     9121 gagaagagca tgcgtgag
//

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