LOCUS       sequence-260641-       13893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive, ectopic expression of human active WRN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13893)
  AUTHORS   Chan EM, Shibue T, McFarland JM, Gaeta B, Ghandi M, Dumont N, 
            Gonzalez A, McPartlan JS, Li T, Zhang Y, Bin Liu J, Lazaro JB, Gu P,
            Piett CG, Apffel A, Ali SO, Deasy R, Keskula P, Ng RWS, Roberts EA, 
            Reznichenko E, Leung L, Alimova M, Schenone M, Islam M, Maruvka YE, 
            Liu Y, Roper J, Raghavan S, Giannakis M, Tseng YY, Nagel ZD, 
            D'Andrea A, Root DE, Boehm JS, Getz G, Chang S, Golub TR, Tsherniak 
            A, Vazquez F, Bass AJ
  TITLE     WRN helicase is a synthetic lethal target in microsatellite unstable
            cancers.
  JOURNAL   Nature. 2019 Apr 10. pii: 10.1038/s41586-019-1102-x. doi: 
            10.1038/s41586-019-1102-x.
  PUBMED    30971823
REFERENCE   2  (bases 1 to 13893)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2019 Apr 10. pii: 10.1038/s41586-019-1102-x. doi: 
            10.1038/s41586-019-1102-x."
COMMENT     Sequence Label: pLX209-neo-active WRN
FEATURES             Location/Qualifiers
     source          1..13893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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"
     promoter        1483..1983
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1902..1920
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1998..2792
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDRLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2052..2071)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2662..2681
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    2840..2957
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3096..4274
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3327..4265
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4222..4242
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      4282..4291
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4312..4341
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4351..4380
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4390..4419
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4429..4458
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4468..4497
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    9519..10107
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9572..9592)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9990..10001)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10179..10412
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10490..10611
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10527..10546)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10581..10600
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10651..10786
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10806..10825)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10807..10825)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10835..10852)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10835..10851)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10844..10866)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10993..11448
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11080..11099)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11290..11311
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11474..11578
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11579..11647,11648..12439)
                     /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     complement(11797..11816)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12610..13198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13099..13118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13352..13369
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13486..13507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13522..13527,13528..13545,13546..13552)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13560..13576
                     /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)."
     primer_bind     13565..13587
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13584..13600
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13584..13600
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     13619..13639
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        13621..13639
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        13667..13893
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccttgcgcct tttccaaggc
     1501 agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa acgcagcggc
     1561 gccgaccctg ggtctcgcac attcttcacg tccgttcgca gcgtcacccg gatcttcgcc
     1621 gctacccttg tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc
     1681 ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg cacgtctcac tagtaccctc
     1741 gcagacggac agcgccaggg agcaatggca gcgcgccgac cgcgatgggc tgtggccaat
     1801 agcggctgct cagcagggcg cgccgagagc agcggccggg aaggggcggt gcgggaggcg
     1861 gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag
     1921 cctccggagc gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga cctctctccc
     1981 cagggggtac cgccaccatg attgaacaag atagattgca cgcaggttct ccggccgctt
     2041 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2101 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2161 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2221 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     2281 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     2341 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     2401 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     2461 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     2521 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     2581 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     2641 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     2701 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     2761 ccttctatcg ccttcttgac gagttcttct gaggtggcga attctagatc ttgagacaaa
     2821 tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg
     2881 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
     2941 caaaaattca aaattttcgg gtttattaca gggacagcag agatccactt tggcgccggc
     3001 tcgagggggc ccgggtgcaa agatggataa agttttaaac agagaggaat ctttgcagct
     3061 aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag
     3121 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
     3181 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     3241 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     3301 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
     3361 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga
     3421 ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag
     3481 gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc
     3541 gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
     3601 atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc
     3661 aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt
     3721 cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg
     3781 ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc
     3841 cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
     3901 ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg
     3961 gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact
     4021 ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt
     4081 cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg
     4141 agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
     4201 agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat
     4261 ttcaggtgtc gtgagggatc cgccaccatg gaaaatcgat ttaaagctat ggagcaaaag
     4321 ctcatttctg aagaggactt gaatgaaatg gagcaaaagc tcatttctga agaggacttg
     4381 aatgaaatgg agcaaaagct catttctgaa gaggacttga atgaaatgga gcaaaagctc
     4441 atttctgaag aggacttgaa tgaaatggag caaaagctca tttctgaaga ggacttgaat
     4501 gaaatggaga gcttgggcga cctcaccatg ggtgaaaaaa aattggaaac aactgcacag
     4561 cagcgaaaat gtcctgaatg gatgaatgtg cagaataaaa gatgtgctgt agaagaaaga
     4621 aaggcatgtg ttcggaagag tgtttttgaa gatgacctcc ccttcttaga attcactgga
     4681 tccattgtgt atagttacga tgctagtgat tgctctttcc tgtcagaaga tattagcatg
     4741 agtctatcag atggggatgt ggtgggattt gacatggagt ggccaccatt atacaacaga
     4801 gggaaacttg gcaaagttgc actaattcag ttgtgtgttt ctgagagcaa atgttacttg
     4861 ttccacgttt cttccatgtc agtttttccc cagggattaa aaatgttgct tgaaaataaa
     4921 gcagttaaaa aggcaggtgt aggaattgaa ggagatcagt ggaaacttct acgagacttt
     4981 gatatcaaat tgaagaattt tgtggagttg acagatgttg ccaataaaaa gctgaaatgt
     5041 acagagacct ggagccttaa cagtctggtt aaacacctct taggtaaaca gctcctgaaa
     5101 gacaagtcta tccgctgtag caattggagt aaatttcctc tcactgagga ccagaaactg
     5161 tatgcagcca ctgatgctta tgctggtttt attatttacc gaaatttaga gattttggat
     5221 gatactgtgc aaaggtttgc tataaataaa gaggaagaaa tcctacttag cgacatgaac
     5281 aaacagttga cttcaatctc tgaggaagtg atggatctgg ctaagcatct tcctcatgct
     5341 ttcagtaaat tggaaaaccc acggagggtt tctatcttac taaaggatat ttcagaaaat
     5401 ctatattcac tgaggaggat gataattggg tctactaaca ttgagactga actgaggccc
     5461 agcaataatt taaacttatt atcctttgaa gattcaacta ctgggggagt acaacagaaa
     5521 caaattagag aacatgaagt tttaattcac gttgaagatg aaacatggga cccaacactt
     5581 gatcatttag ctaaacatga tggagaagat gtacttggaa ataaagtgga acgaaaagaa
     5641 gatggatttg aagatggagt agaagacaac aaattgaaag agaatatgga aagagcttgt
     5701 ttgatgtcgt tagatattac agaacatgaa ctccaaattt tggaacagca gtctcaggaa
     5761 gaatatctta gtgatattgc ttataaatct actgagcatt tatctcccaa tgataatgaa
     5821 aacgatacgt cctatgtaat tgagagtgat gaagatttag aaatggagat gcttaagcat
     5881 ttatctccca atgataatga aaacgatacg tcctatgtaa ttgagagtga tgaagattta
     5941 gaaatggaga tgcttaagtc tttagaaaac ctcaatagtg gcacggtaga accaactcat
     6001 tctaaatgct taaaaatgga aagaaatctg ggtcttccta ctaaagaaga agaagaagat
     6061 gatgaaaatg aagctaatga aggggaagaa gatgatgata aggacttttt gtggccagca
     6121 cccaatgaag agcaagttac ttgcctcaag atgtactttg gccattccag ttttaaacca
     6181 gttcagtgga aagtgattca ttcagtatta gaagaaagaa gagataatgt tgctgtcatg
     6241 gcaactggat atggaaagag tttgtgcttc cagtatccac ctgtttatgt aggcaagatt
     6301 ggccttgtta tctctcccct tatttctctg atggaagacc aagtgctaca gcttaaaatg
     6361 tccaacatcc cagcttgctt ccttggatca gcacagtcag aaaatgttct aacagatatt
     6421 aaattaggta aataccggat tgtatacgta actccagaat actgttcagg taacatgggc
     6481 ctgctccagc aacttgaggc tgatattggt atcacgctca ttgctgtgga tgaggctcac
     6541 tgtatttctg agtgggggca tgattttagg gattcattca ggaagttggg ctccctaaag
     6601 acagcactgc caatggttcc aatcgttgca cttactgcta ctgcaagttc ttcaatccgg
     6661 gaagacattg tacgttgctt aaatctgaga aatcctcaga tcacctgtac tggttttgat
     6721 cgaccaaacc tgtatttaga agttaggcga aaaacaggga atatccttca ggatctgcag
     6781 ccatttcttg tcaaaacaag ttcccactgg gaatttgaag gtccaacaat catctactgt
     6841 ccttctagaa aaatgacaca acaagttaca ggtgaactta ggaaactgaa tctatcctgt
     6901 ggaacatacc atgcgggcat gagttttagc acaaggaaag acattcatca taggtttgta
     6961 agagatgaaa ttcagtgtgt catagctacc atagcttttg gaatgggcat taataaagct
     7021 gacattcgcc aagtcattca ttacggtgct cctaaggaca tggaatcata ttatcaggag
     7081 attggtagag ctggtcgtga tggacttcaa agttcttgtc acgtcctctg ggctcctgca
     7141 gacattaact taaataggca ccttcttact gagatacgta atgagaagtt tcgattatac
     7201 aaattaaaga tgatggcaaa gatggaaaaa tatcttcatt ctagcagatg taggagacaa
     7261 atcatcttgt ctcattttga ggacaaacaa gtacaaaaag cctccttggg aattatggga
     7321 actgaaaaat gctgtgataa ttgcaggtcc agattggatc attgctattc catggatgac
     7381 tcagaggata catcctggga ctttggtcca caagcattta agcttttgtc tgctgtggac
     7441 atcttaggcg aaaaatttgg aattgggctt ccaattttat ttctccgagg atctaattct
     7501 cagcgtcttg ccgatcaata tcgcaggcac agtttatttg gcactggcaa ggatcaaaca
     7561 gagagttggt ggaaggcttt ttcccgtcag ctgatcactg agggattctt ggtagaagtt
     7621 tctcggtata acaaatttat gaagatttgc gcccttacga aaaagggtag aaattggctt
     7681 cataaagcta atacagaatc tcagagcctc atccttcaag ctaatgaaga attgtgtcca
     7741 aagaagttgc ttctgcctag ttcgaaaact gtatcttcgg gcaccaaaga gcattgttat
     7801 aatcaagtac cagttgaatt aagtacagag aagaagtcta acttggagaa gttatattct
     7861 tataaaccat gtgataagat ttcttctggg agtaacattt ctaaaaaaag tatcatggta
     7921 cagtcaccag aaaaagctta cagttcctca cagcctgtta tttcggcaca agagcaggag
     7981 actcagattg tgttatatgg caaattggta gaagctaggc agaaacatgc caataaaatg
     8041 gatgttcccc cagctattct ggcaacaaac aagatactgg tggatatggc caaaatgaga
     8101 ccaactacgg ttgaaaacgt aaaaaggatt gatggtgttt ctgaaggcaa agctgccatg
     8161 ttggcccctc tgttggaagt catcaaacat ttctgccaaa caaatagtgt tcagacagac
     8221 ctcttttcaa gtacaaaacc tcaagaagaa cagaagacga gtctggtagc aaaaaataaa
     8281 atatgcacac tttcacagtc tatggccatc acatactctt tattccaaga aaagaagatg
     8341 cctttgaaga gcatagctga gagcaggatt ctgcctctca tgacaattgg catgcactta
     8401 tcccaagcgg tgaaagctgg ctgccccctt gatttggagc gagcaggcct gactccagag
     8461 gttcagaaga ttattgctga tgttatccga aaccctcccg tcaactcaga tatgagtaaa
     8521 attagcctaa tcagaatgtt agttcctgaa aacattgaca cgtaccttat ccacatggca
     8581 attgagatcc ttaaacatgg tcctgacagt ggacttcaac cttcatgtga tgtcaacaaa
     8641 aggagatgtt ttcccggttc tgaagagatc tgttcaagtt ctaagagaag caaggaagaa
     8701 gtaggcatca atactgagac ttcatctgca gagagaaaga gacgattacc tgtgtggttt
     8761 gccaaaggaa gtgataccag caagaaatta atggacaaaa cgaaaagggg aggtcttttt
     8821 agttaagctg gcaattacca gaacaattat gtttcttgct gtattataag aggatagcta
     8881 tattttattt ctgaagagta aggagtagta ttttggctta aaaatcattc taattacaaa
     8941 gttcactgtt tattgaagaa ctggcatctt aaatcagcct tccgcaattc atgtagtttc
     9001 tgggtcttct gggagcctac gtgagtacat cacctaacag aatattaaat tagacttcct
     9061 gtaagattgc tttaagaaac tgttactgtc ctgttttcta atctctttat taaaacagtg
     9121 tatttggaaa atgttatgtg ctctgatttg atatagataa cagattagta gttacatggt
     9181 aattatgtga tataaaatat tcatatatta tcaaaattct gttttgtaaa tgtaagaaag
     9241 catagttatt ttacaaattg tttttactgt cttttgaaga agttcttaaa tacgttgtta
     9301 aatggtatta gttgaccagg gcagtgaaaa tgaaaccgca ttttgggtgc cattaaatag
     9361 ggaaaaaaca tgtaaaaaat gtaaaatgga gaccaattgc actaggcaag tgtatatttt
     9421 gtattttata tacaatttct attatttttc aagtaataaa acaatgtttt tcatnaaaaa
     9481 aaaaaaaaaa aaaaaactcg acgacgcgtt aagtcgacaa tcaacctctg gattacaaaa
     9541 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     9601 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     9661 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     9721 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     9781 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     9841 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     9901 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     9961 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
    10021 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
    10081 ggatctccct ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc
    10141 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
    10201 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
    10261 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
    10321 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
    10381 acccttttag tcagtgtgga aaatctctag cagtacgtat agtagttcat gtcatcttat
    10441 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat
    10501 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
    10561 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
    10621 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
    10681 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
    10741 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca
    10801 attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg
    10861 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
    10921 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
    10981 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
    11041 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
    11101 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
    11161 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
    11221 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
    11281 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
    11341 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
    11401 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
    11461 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
    11521 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
    11581 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
    11641 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
    11701 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
    11761 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
    11821 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
    11881 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
    11941 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
    12001 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
    12061 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
    12121 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
    12181 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
    12241 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
    12301 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
    12361 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
    12421 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
    12481 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
    12541 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
    12601 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
    12661 accgctacca gcggtggttt gtttgccgga tcaagagcca ccaactcttt ttccgaaggt
    12721 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
    12781 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
    12841 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
    12901 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
    12961 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
    13021 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
    13081 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
    13141 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
    13201 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
    13261 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
    13321 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
    13381 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
    13441 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
    13501 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
    13561 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc
    13621 aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttaatgta gtcttatgca
    13681 atactcttgt agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag
    13741 aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga
    13801 aggcaacaga cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat
    13861 attgtattta agtgcctagc tcgatacata aac
//
