pLVX-EF1alpha-SARS-CoV-2-orf9c-IRES-Puro

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载体基本信息

质粒名称: pLVX-EF1alpha-SARS-CoV-2-orf9c-IRES-Puro
出品公司: Addgene
目录编号: 141393
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: 8825bp
完整质粒大小: 9126 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-orf9c
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 (N terminal on insert)
克隆方法: 未知

文献引用方法
材料和方法部分: pLXV-EF1alpha-2xStrep-SARS-CoV-2-orf9c-IRES-Puro was a gift from Nevan Krogan (Addgene plasmid # 141393 ; http://n2t.net/addgene:141393 ; RRID:Addgene_141393)
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-orf9c-IRES-Puro载体图谱

载体简介

载体序列

LOCUS       pLXV-EF1alpha-2xStrep-SARS-CoV-2-orf9c-IRES-Puro                9126 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Lentiviral expression of SARS-CoV-2 protein; transient expression 
            and generate lentivirus.
ACCESSION   .
VERSION     .
KEYWORDS    pLXV-EF1alpha-2xStrep-SARS-CoV-2-orf9c-IRES-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9126)
  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 9126)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9126
                     /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             1216..1239
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1276..1299
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    1539..2112
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1706..1723)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1933..1952
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2145..2744
                     /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(2145..2164)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2641..2661
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2758..3346
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2811..2831)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3229..3240)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3553..4186
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4314..4330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4314..4330)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4327..4349)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4338..4354
                     /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(4362..4392)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4407..4428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4545..4562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4716..5304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4796..4815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5475..6335)
                     /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     6098..6117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6336..6440)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    6488..6622
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6525..6544)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6579..6598
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             6790..7423
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7470..7595
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8092..8325
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8510..8554
                     /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    8817..8932
                     /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 ccatgtggtc ccatccgcag tttgagaagg gtggtggttc aggcggaggc
     1261 tccgggggcg ggagctggtc tcacccgcaa tttgaaaaag gcgctgcggc tcttcaatcc
     1321 tgctataact tcttgaaaga gcaacactgc caaaaggcga gcacacagaa aggagccgag
     1381 gcagccgtaa agccgcttct tgtccctcat cacgttgtcg ccacagtcca agaaatacag
     1441 cttcaagcgg ccgttggcga gcttctgctc ctcgaatggc tcgcaatggc ggtgatgttg
     1501 ttgctgttgt gctgctgtct tacggattga ggatcccgcc cctctccctc ccccccccct
     1561 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     1621 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     1681 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     1741 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     1801 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     1861 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     1921 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     1981 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     2041 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     2101 acacgatgat aagcttgcca caacccacaa ggagacgacc ttccatgacc gagtacaagc
     2161 ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg
     2221 cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg
     2281 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     2341 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     2401 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     2461 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     2521 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     2581 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     2641 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     2701 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctagacgcgt ctggaacaat
     2761 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     2821 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2881 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     2941 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3001 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3061 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3121 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     3181 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3241 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3301 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattaattct
     3361 gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg
     3421 ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc
     3481 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     3541 aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt
     3601 ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg
     3661 tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg
     3721 tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga
     3781 tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc
     3841 acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca
     3901 agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc
     3961 gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta
     4021 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     4081 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     4141 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt
     4201 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggcct
     4261 tgacattgct agcgtttacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     4321 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4381 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4441 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4501 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4561 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4621 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4681 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4741 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4801 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4861 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4921 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4981 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5041 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5101 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5161 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5221 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5281 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5341 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5401 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5461 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5521 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5581 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5641 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5701 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5761 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5821 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5881 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5941 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6001 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6061 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6121 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6181 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6241 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6301 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6361 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6421 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     6481 ggagatcaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     6541 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     6601 tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact
     6661 tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg
     6721 tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa
     6781 cttagtagtt ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga
     6841 tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca
     6901 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag
     6961 aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg
     7021 atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg
     7081 tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg
     7141 gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag
     7201 ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac
     7261 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     7321 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     7381 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
     7441 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
     7501 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
     7561 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
     7621 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
     7681 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
     7741 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     7801 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     7861 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
     7921 ccgcacagca agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     7981 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     8041 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     8101 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     8161 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     8221 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     8281 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     8341 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     8401 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     8461 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     8521 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     8581 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     8641 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     8701 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg
     8761 agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccttta
     8821 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     8881 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     8941 attacaggga cagcagagat ccagtttatc gatgagtaat tcatacaaaa ggactcgccc
     9001 ctgccttggg gaatcccagg gaccgtcgtt aaactcccac taacgtagaa cccagagatc
     9061 gctgcgttcc cgccccctca cccgcccgct ctcgtcatca ctgaggtgga gaagagcatg
     9121 cgtgag
//

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