LOCUS       sequence-245961-       13892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive, ectopic expression of an exonuclease-dead mutant of 
            human WRN (E84A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13892)
  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 13892)
  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-WRN E84A
FEATURES             Location/Qualifiers
     source          1..13892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    47..172
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    665..898
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1083..1127
                     /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        1302..1802
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1721..1739
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1817..2611
                     /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(1871..1890)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2481..2500
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    2659..2776
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2915..4093
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3146..4084
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4041..4061
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      4101..4110
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4131..4160
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4170..4199
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4209..4238
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4248..4277
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4287..4316
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    9337..9925
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9390..9410)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9808..9819)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9997..10230
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10308..10429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10345..10364)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10399..10418
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10469..10604
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10624..10643)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10625..10643)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10653..10670)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10653..10669)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10662..10684)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10811..11266
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10898..10917)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11108..11129
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11292..11396
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11397..11465,11466..12257)
                     /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(11615..11634)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12428..13016
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12917..12936
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13170..13187
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13304..13325
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13340..13345,13346..13363,13364..13370)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13378..13394
                     /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     13383..13405
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13402..13418
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13402..13418
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     13437..13457
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        13439..13457
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        13485..13711
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             13712..13892
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
       61 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
      121 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
      181 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
      241 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
      301 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
      361 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
      421 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
      481 aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag
      541 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
      601 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
      661 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
      721 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
      781 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
      841 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
      901 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
      961 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     1021 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     1081 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     1141 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     1201 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     1261 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cttgcgcctt ttccaaggca
     1321 gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg
     1381 ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg
     1441 ctacccttgt gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct
     1501 tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg
     1561 cagacggaca gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata
     1621 gcggctgctc agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg
     1681 ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc
     1741 ctccggagcg cacgtcggca gtcggctccc tcgttgaccg aatcaccgac ctctctcccc
     1801 agggggtacc gccaccatga ttgaacaaga tagattgcac gcaggttctc cggccgcttg
     1861 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     1921 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     1981 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     2041 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     2101 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     2161 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     2221 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     2281 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     2341 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     2401 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     2461 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     2521 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     2581 cttctatcgc cttcttgacg agttcttctg aggtggcgaa ttctagatct tgagacaaat
     2641 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
     2701 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
     2761 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccacttt ggcgccggct
     2821 cgagggggcc cgggtgcaaa gatggataaa gttttaaaca gagaggaatc tttgcagcta
     2881 atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga
     2941 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc
     3001 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     3061 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     3121 caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct
     3181 ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat
     3241 tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg
     3301 agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg
     3361 aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa
     3421 tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca
     3481 agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc
     3541 ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg
     3601 gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc
     3661 gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct
     3721 tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg
     3781 tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc
     3841 cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt ggagtacgtc
     3901 gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga
     3961 gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga
     4021 gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt
     4081 tcaggtgtcg tgagggatcc gccaccatgg aaaatcgatt taaagctatg gagcaaaagc
     4141 tcatttctga agaggacttg aatgaaatgg agcaaaagct catttctgaa gaggacttga
     4201 atgaaatgga gcaaaagctc atttctgaag aggacttgaa tgaaatggag caaaagctca
     4261 tttctgaaga ggacttgaat gaaatggagc aaaagctcat ttctgaagag gacttgaatg
     4321 aaatggagag cttgggcgac ctcaccatgg gtgaaaaaaa attggaaaca actgcacagc
     4381 agcgaaaatg tcctgaatgg atgaatgtgc agaataaaag atgtgctgta gaagaaagaa
     4441 aggcatgtgt tcggaagagt gtttttgaag atgacctccc cttcttagaa ttcactggat
     4501 ccattgtgta tagttacgat gctagtgatt gctctttcct gtcagaagat attagcatga
     4561 gtctatcaga tggggatgtg gtgggatttg acatggcgtg gcctccatta tacaacagag
     4621 ggaaacttgg caaagttgca ctaattcagt tgtgtgtttc tgagagcaaa tgttacttgt
     4681 tccacgtttc ttccatgtca gtttttcccc agggattaaa aatgttgctt gaaaataaag
     4741 cagttaaaaa ggcaggtgta ggaattgaag gagatcagtg gaaacttcta cgagactttg
     4801 atatcaaatt gaagaatttt gtggagttga cagatgttgc caataaaaag ctgaaatgta
     4861 cagagacctg gagccttaac agtctggtta aacacctctt aggtaaacag ctcctgaaag
     4921 acaagtctat ccgctgtagc aattggagta aatttcctct cactgaggac cagaaactgt
     4981 atgcagccac tgatgcttat gctggtttta ttatttaccg aaatttagag attttggatg
     5041 atactgtgca aaggtttgct ataaataaag aggaagaaat cctacttagc gacatgaaca
     5101 aacagttgac ttcaatctct gaggaagtga tggatctggc taagcatctt cctcatgctt
     5161 tcagtaaatt ggaaaaccca cggagggttt ctatcttact aaaggatatt tcagaaaatc
     5221 tatattcact gaggaggatg ataattgggt ctactaacat tgagactgaa ctgaggccca
     5281 gcaataattt aaacttatta tcctttgaag attcaactac tgggggagta caacagaaac
     5341 aaattagaga acatgaagtt ttaattcacg ttgaagatga aacatgggac ccaacacttg
     5401 atcatttagc taaacatgat ggagaagatg tacttggaaa taaagtggaa cgaaaagaag
     5461 atggatttga agatggagta gaagacaaca aattgaaaga gaatatggaa agagcttgtt
     5521 tgatgtcgtt agatattaca gaacatgaac tccaaatttt ggaacagcag tctcaggaag
     5581 aatatcttag tgatattgct tataaatcta ctgagcattt atctcccaat gataatgaaa
     5641 acgatacgtc ctatgtaatt gagagtgatg aagatttaga aatggagatg cttaagcatt
     5701 tatctcccaa tgataatgaa aacgatacgt cctatgtaat tgagagtgat gaagatttag
     5761 aaatggagat gcttaagtct ttagaaaacc tcaatagtgg cacggtagaa ccaactcatt
     5821 ctaaatgctt aaaaatggaa agaaatctgg gtcttcctac taaagaagaa gaagaagatg
     5881 atgaaaatga agctaatgaa ggggaagaag atgatgataa ggactttttg tggccagcac
     5941 ccaatgaaga gcaagttact tgcctcaaga tgtactttgg ccattccagt tttaaaccag
     6001 ttcagtggaa agtgattcat tcagtattag aagaaagaag agataatgtt gctgtcatgg
     6061 caactggata tggaaagagt ttgtgcttcc agtatccacc tgtttatgta ggcaagattg
     6121 gccttgttat ctctcccctt atttctctga tggaagacca agtgctacag cttaaaatgt
     6181 ccaacatccc agcttgcttc cttggatcag cacagtcaga aaatgttcta acagatatta
     6241 aattaggtaa ataccggatt gtatacgtaa ctccagaata ctgttcaggt aacatgggcc
     6301 tgctccagca acttgaggct gatattggta tcacgctcat tgctgtggat gaggctcact
     6361 gtatttctga gtgggggcat gattttaggg attcattcag gaagttgggc tccctaaaga
     6421 cagcactgcc aatggttcca atcgttgcac ttactgctac tgcaagttct tcaatccggg
     6481 aagacattgt acgttgctta aatctgagaa atcctcagat cacctgtact ggttttgatc
     6541 gaccaaacct gtatttagaa gttaggcgaa aaacagggaa tatccttcag gatctgcagc
     6601 catttcttgt caaaacaagt tcccactggg aatttgaagg tccaacaatc atctactgtc
     6661 cttctagaaa aatgacacaa caagttacag gtgaacttag gaaactgaat ctatcctgtg
     6721 gaacatacca tgcgggcatg agttttagca caaggaaaga cattcatcat aggtttgtaa
     6781 gagatgaaat tcagtgtgtc atagctacca tagcttttgg aatgggcatt aataaagctg
     6841 acattcgcca agtcattcat tacggtgctc ctaaggacat ggaatcatat tatcaggaga
     6901 ttggtagagc tggtcgtgat ggacttcaaa gttcttgtca cgtcctctgg gctcctgcag
     6961 acattaactt aaataggcac cttcttactg agatacgtaa tgagaagttt cgattataca
     7021 aattaaagat gatggcaaag atggaaaaat atcttcattc tagcagatgt aggagacaaa
     7081 tcatcttgtc tcattttgag gacaaacaag tacaaaaagc ctccttggga attatgggaa
     7141 ctgaaaaatg ctgtgataat tgcaggtcca gattggatca ttgctattcc atggatgact
     7201 cagaggatac atcctgggac tttggtccac aagcatttaa gcttttgtct gctgtggaca
     7261 tcttaggcga aaaatttgga attgggcttc caattttatt tctccgagga tctaattctc
     7321 agcgtcttgc cgatcaatat cgcaggcaca gtttatttgg cactggcaag gatcaaacag
     7381 agagttggtg gaaggctttt tcccgtcagc tgatcactga gggattcttg gtagaagttt
     7441 ctcggtataa caaatttatg aagatttgcg cccttacgaa aaagggtaga aattggcttc
     7501 ataaagctaa tacagaatct cagagcctca tccttcaagc taatgaagaa ttgtgtccaa
     7561 agaagttgct tctgcctagt tcgaaaactg tatcttcggg caccaaagag cattgttata
     7621 atcaagtacc agttgaatta agtacagaga agaagtctaa cttggagaag ttatattctt
     7681 ataaaccatg tgataagatt tcttctggga gtaacatttc taaaaaaagt atcatggtac
     7741 agtcaccaga aaaagcttac agttcctcac agcctgttat ttcggcacaa gagcaggaga
     7801 ctcagattgt gttatatggc aaattggtag aagctaggca gaaacatgcc aataaaatgg
     7861 atgttccccc agctattctg gcaacaaaca agatactggt ggatatggcc aaaatgagac
     7921 caactacggt tgaaaacgta aaaaggattg atggtgtttc tgaaggcaaa gctgccatgt
     7981 tggcccctct gttggaagtc atcaaacatt tctgccaaac aaatagtgtt cagacagacc
     8041 tcttttcaag tacaaaacct caagaagaac agaagacgag tctggtagca aaaaataaaa
     8101 tatgcacact ttcacagtct atggccatca catactcttt attccaagaa aagaagatgc
     8161 ctttgaagag catagctgag agcaggattc tgcctctcat gacaattggc atgcacttat
     8221 cccaagcggt gaaagctggc tgcccccttg atttggagcg agcaggcctg actccagagg
     8281 ttcagaagat tattgctgat gttatccgaa accctcccgt caactcagat atgagtaaaa
     8341 ttagcctaat cagaatgtta gttcctgaaa acattgacac gtaccttatc cacatggcaa
     8401 ttgagatcct taaacatggt cctgacagtg gacttcaacc ttcatgtgat gtcaacaaaa
     8461 ggagatgttt tcccggttct gaagagatct gttcaagttc taagagaagc aaggaagaag
     8521 taggcatcaa tactgagact tcatctgcag agagaaagag acgattacct gtgtggtttg
     8581 ccaaaggaag tgataccagc aagaaattaa tggacaaaac gaaaagggga ggtcttttta
     8641 gttaagctgg caattaccag aacaattatg tttcttgctg tattataaga ggatagctat
     8701 attttatttc tgaagagtaa ggagtagtat tttggcttaa aaatcattct aattacaaag
     8761 ttcactgttt attgaagaac tggcatctta aatcagcctt ccgcaattca tgtagtttct
     8821 gggtcttctg ggagcctacg tgagtacatc acctaacaga atattaaatt agacttcctg
     8881 taagattgct ttaagaaact gttactgtcc tgttttctaa tctctttatt aaaacagtgt
     8941 atttggaaaa tgttatgtgc tctgatttga tatagataac agattagtag ttacatggta
     9001 attatgtgat ataaaatatt catatattat caaaattctg ttttgtaaat gtaagaaagc
     9061 atagttattt tacaaattgt ttttactgtc ttttgaagaa gttcttaaat acgttgttaa
     9121 atggtattag ttgaccaggg cagtgaaaat gaaaccgcat tttgggtgcc attaaatagg
     9181 gaaaaaacat gtaaaaaatg taaaatggag accaattgca ctaggcaagt gtatattttg
     9241 tattttatat acaatttcta ttatttttca agtaataaaa caatgttttt cataaaaaaa
     9301 aaaaaaaaaa aaaactcgac gacgcgttaa gtcgacaatc aacctctgga ttacaaaatt
     9361 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     9421 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     9481 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     9541 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     9601 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     9661 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     9721 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     9781 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     9841 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     9901 atctcccttt gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg
     9961 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
    10021 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
    10081 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
    10141 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
    10201 ccttttagtc agtgtggaaa atctctagca gtacgtatag tagttcatgt catcttatta
    10261 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
    10321 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
    10381 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
    10441 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
    10501 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
    10561 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat
    10621 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
    10681 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
    10741 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
    10801 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
    10861 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
    10921 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
    10981 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
    11041 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
    11101 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
    11161 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
    11221 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
    11281 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
    11341 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
    11401 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
    11461 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
    11521 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
    11581 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
    11641 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
    11701 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
    11761 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
    11821 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
    11881 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
    11941 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
    12001 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
    12061 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
    12121 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
    12181 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
    12241 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
    12301 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
    12361 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
    12421 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
    12481 cgctaccagc ggtggtttgt ttgccggatc aagagccacc aactcttttt ccgaaggtaa
    12541 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
    12601 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
    12661 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
    12721 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
    12781 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
    12841 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
    12901 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
    12961 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
    13021 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
    13081 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
    13141 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
    13201 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
    13261 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
    13321 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
    13381 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
    13441 ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat
    13501 actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa
    13561 aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag
    13621 gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat
    13681 tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc
    13741 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
    13801 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
    13861 acccttttag tcagtgtgga aaatctctag ca
//
