LOCUS       sequence-268910-       13893 bp    DNA     circular SYN 14-APR-2025
DEFINITION  constitutive, ectopic expression of a helicase-dead mutant of human 
            WRN (K577M).
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 Apr 14, 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 K577M
FEATURES             Location/Qualifiers
     source          1..13893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1187..1196
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1217..1246
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1256..1285
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1295..1324
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1334..1363
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1373..1402
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    6424..7012
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6477..6497)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6895..6906)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7084..7317
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7395..7516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7432..7451)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7486..7505
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7556..7691
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7711..7730)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7712..7730)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7740..7757)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7740..7756)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7749..7771)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7898..8353
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7985..8004)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8195..8216
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8379..8483
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8484..8552,8553..9344)
                     /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(8702..8721)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9515..10103
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10004..10023
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10257..10274
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10391..10412
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10427..10432,10433..10450,10451..10457)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10465..10481
                     /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     10470..10492
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10489..10505
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10489..10505
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10524..10544
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10526..10544
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        10572..10798
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             10799..10979
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11026..11151
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11644..11877
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12062..12106
                     /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        12281..12781
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     12700..12718
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             12796..13590
                     /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(12850..12869)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     13460..13479
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    13638..13755
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag ggatccgcca ccatggaaaa
     1201 tcgatttaaa gctatggagc aaaagctcat ttctgaagag gacttgaatg aaatggagca
     1261 aaagctcatt tctgaagagg acttgaatga aatggagcaa aagctcattt ctgaagagga
     1321 cttgaatgaa atggagcaaa agctcatttc tgaagaggac ttgaatgaaa tggagcaaaa
     1381 gctcatttct gaagaggact tgaatgaaat ggagagcttg ggcgacctca ccatgggtga
     1441 aaaaaaattg gaaacaactg cacagcagcg aaaatgtcct gaatggatga atgtgcagaa
     1501 taaaagatgt gctgtagaag aaagaaaggc atgtgttcgg aagagtgttt ttgaagatga
     1561 cctccccttc ttagaattca ctggatccat tgtgtatagt tacgatgcta gtgattgctc
     1621 tttcctgtca gaagatatta gcatgagtct atcagatggg gatgtggtgg gatttgacat
     1681 ggagtggcca ccattataca acagagggaa acttggcaaa gttgcactaa ttcagttgtg
     1741 tgtttctgag agcaaatgtt acttgttcca cgtttcttcc atgtcagttt ttccccaggg
     1801 attaaaaatg ttgcttgaaa ataaagcagt taaaaaggca ggtgtaggaa ttgaaggaga
     1861 tcagtggaaa cttctacgag actttgatat caaattgaag aattttgtgg agttgacaga
     1921 tgttgccaat aaaaagctga aatgtacaga gacctggagc cttaacagtc tggttaaaca
     1981 cctcttaggt aaacagctcc tgaaagacaa gtctatccgc tgtagcaatt ggagtaaatt
     2041 tcctctcact gaggaccaga aactgtatgc agccactgat gcttatgctg gttttattat
     2101 ttaccgaaat ttagagattt tggatgatac tgtgcaaagg tttgctataa ataaagagga
     2161 agaaatccta cttagcgaca tgaacaaaca gttgacttca atctctgagg aagtgatgga
     2221 tctggctaag catcttcctc atgctttcag taaattggaa aacccacgga gggtttctat
     2281 cttactaaag gatatttcag aaaatctata ttcactgagg aggatgataa ttgggtctac
     2341 taacattgag actgaactga ggcccagcaa taatttaaac ttattatcct ttgaagattc
     2401 aactactggg ggagtacaac agaaacaaat tagagaacat gaagttttaa ttcacgttga
     2461 agatgaaaca tgggacccaa cacttgatca tttagctaaa catgatggag aagatgtact
     2521 tggaaataaa gtggaacgaa aagaagatgg atttgaagat ggagtagaag acaacaaatt
     2581 gaaagagaat atggaaagag cttgtttgat gtcgttagat attacagaac atgaactcca
     2641 aattttggaa cagcagtctc aggaagaata tcttagtgat attgcttata aatctactga
     2701 gcatttatct cccaatgata atgaaaacga tacgtcctat gtaattgaga gtgatgaaga
     2761 tttagaaatg gagatgctta agcatttatc tcccaatgat aatgaaaacg atacgtccta
     2821 tgtaattgag agtgatgaag atttagaaat ggagatgctt aagtctttag aaaacctcaa
     2881 tagtggcacg gtagaaccaa ctcattctaa atgcttaaaa atggaaagaa atctgggtct
     2941 tcctactaaa gaagaagaag aagatgatga aaatgaagct aatgaagggg aagaagatga
     3001 tgataaggac tttttgtggc cagcacccaa tgaagagcaa gttacttgcc tcaagatgta
     3061 ctttggccat tccagtttta aaccagttca gtggaaagtg attcattcag tattagaaga
     3121 aagaagagat aatgttgctg tcatggcaac tggatatgga atgagtttgt gcttccagta
     3181 tccacctgtt tatgtaggca agattggcct tgttatctct ccccttattt ctctgatgga
     3241 agaccaagtg ctacagctta aaatgtccaa catcccagct tgcttccttg gatcagcaca
     3301 gtcagaaaat gttctaacag atattaaatt aggtaaatac cggattgtat acgtaactcc
     3361 agaatactgt tcaggtaaca tgggcctgct ccagcaactt gaggctgata ttggtatcac
     3421 gctcattgct gtggatgagg ctcactgtat ttctgagtgg gggcatgatt ttagggattc
     3481 attcaggaag ttgggctccc taaagacagc actgccaatg gttccaatcg ttgcacttac
     3541 tgctactgca agttcttcaa tccgggaaga cattgtacgt tgcttaaatc tgagaaatcc
     3601 tcagatcacc tgtactggtt ttgatcgacc aaacctgtat ttagaagtta ggcgaaaaac
     3661 agggaatatc cttcaggatc tgcagccatt tcttgtcaaa acaagttccc actgggaatt
     3721 tgaaggtcca acaatcatct actgtccttc tagaaaaatg acacaacaag ttacaggtga
     3781 acttaggaaa ctgaatctat cctgtggaac ataccatgcg ggcatgagtt ttagcacaag
     3841 gaaagacatt catcataggt ttgtaagaga tgaaattcag tgtgtcatag ctaccatagc
     3901 ttttggaatg ggcattaata aagctgacat tcgccaagtc attcattacg gtgctcctaa
     3961 ggacatggaa tcatattatc aggagattgg tagagctggt cgtgatggac ttcaaagttc
     4021 ttgtcacgtc ctctgggctc ctgcagacat taacttaaat aggcaccttc ttactgagat
     4081 acgtaatgag aagtttcgat tatacaaatt aaagatgatg gcaaagatgg aaaaatatct
     4141 tcattctagc agatgtagga gacaaatcat cttgtctcat tttgaggaca aacaagtaca
     4201 aaaagcctcc ttgggaatta tgggaactga aaaatgctgt gataattgca ggtccagatt
     4261 ggatcattgc tattccatgg atgactcaga ggatacatcc tgggactttg gtccacaagc
     4321 atttaagctt ttgtctgctg tggacatctt aggcgaaaaa tttggaattg ggcttccaat
     4381 tttatttctc cgaggatcta attctcagcg tcttgccgat caatatcgca ggcacagttt
     4441 atttggcact ggcaaggatc aaacagagag ttggtggaag gctttttccc gtcagctgat
     4501 cactgaggga ttcttggtag aagtttctcg gtataacaaa tttatgaaga tttgcgccct
     4561 tacgaaaaag ggtagaaatt ggcttcataa agctaataca gaatctcaga gcctcatcct
     4621 tcaagctaat gaagaattgt gtccaaagaa gttgcttctg cctagttcga aaactgtatc
     4681 ttcgggcacc aaagagcatt gttataatca agtaccagtt gaattaagta cagagaagaa
     4741 gtctaacttg gagaagttat attcttataa accatgtgat aagatttctt ctgggagtaa
     4801 catttctaaa aaaagtatca tggtacagtc accagaaaaa gcttacagtt cctcacagcc
     4861 tgttatttcg gcacaagagc aggagactca gattgtgtta tatggcaaat tggtagaagc
     4921 taggcagaaa catgccaata aaatggatgt tcccccagct attctggcaa caaacaagat
     4981 actggtggat atggccaaaa tgagaccaac tacggttgaa aacgtaaaaa ggattgatgg
     5041 tgtttctgaa ggcaaagctg ccatgttggc ccctctgttg gaagtcatca aacatttctg
     5101 ccaaacaaat agtgttcaga cagacctctt ttcaagtaca aaacctcaag aagaacagaa
     5161 gacgagtctg gtagcaaaaa ataaaatatg cacactttca cagtctatgg ccatcacata
     5221 ctctttattc caagaaaaga agatgccttt gaagagcata gctgagagca ggattctgcc
     5281 tctcatgaca attggcatgc acttatccca agcggtgaaa gctggctgcc cccttgattt
     5341 ggagcgagca ggcctgactc cagaggttca gaagattatt gctgatgtta tccgaaaccc
     5401 tcccgtcaac tcagatatga gtaaaattag cctaatcaga atgttagttc ctgaaaacat
     5461 tgacacgtac cttatccaca tggcaattga gatccttaaa catggtcctg acagtggact
     5521 tcaaccttca tgtgatgtca acaaaaggag atgttttccc ggttctgaag agatctgttc
     5581 aagttctaag agaagcaagg aagaagtagg catcaatact gagacttcat ctgcagagag
     5641 aaagagacga ttacctgtgt ggtttgccaa aggaagtgat accagcaaga aattaatgga
     5701 caaaacgaaa aggggaggtc tttttagtta agctggcaat taccagaaca attatgtttc
     5761 ttgctgtatt ataagaggat agctatattt tatttctgaa gagtaaggag tagtattttg
     5821 gcttaaaaat cattctaatt acaaagttca ctgtttattg aagaactggc atcttaaatc
     5881 agccttccgc aattcatgta gtttctgggt cttctgggag cctacgtgag tacatcacct
     5941 aacagaatat taaattagac ttcctgtaag attgctttaa gaaactgtta ctgtcctgtt
     6001 ttctaatctc tttattaaaa cagtgtattt ggaaaatgtt atgtgctctg atttgatata
     6061 gataacagat tagtagttac atggtaatta tgtgatataa aatattcata tattatcaaa
     6121 attctgtttt gtaaatgtaa gaaagcatag ttattttaca aattgttttt actgtctttt
     6181 gaagaagttc ttaaatacgt tgttaaatgg tattagttga ccagggcagt gaaaatgaaa
     6241 ccgcattttg ggtgccatta aatagggaaa aaacatgtaa aaaatgtaaa atggagacca
     6301 attgcactag gcaagtgtat attttgtatt ttatatacaa tttctattat ttttcaagta
     6361 ataaaacaat gtttttcata aaaaaaaaaa aaaaaaaaaa nctcgacgac gcgttaagtc
     6421 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     6481 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     6541 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     6601 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     6661 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     6721 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     6781 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     6841 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     6901 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     6961 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcgtcgactt
     7021 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     7081 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     7141 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     7201 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     7261 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     7321 cgtatagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     7381 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     7441 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     7501 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc
     7561 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     7621 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     7681 ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     7741 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     7801 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     7861 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     7921 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     7981 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     8041 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     8101 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     8161 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     8221 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     8281 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     8341 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     8401 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     8461 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     8521 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     8581 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     8641 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     8701 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     8761 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     8821 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     8881 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     8941 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     9001 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     9061 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     9121 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     9181 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     9241 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     9301 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     9361 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     9421 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     9481 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     9541 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     9601 agccaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     9661 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     9721 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     9781 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     9841 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     9901 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     9961 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
    10021 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
    10081 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
    10141 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
    10201 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
    10261 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
    10321 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
    10381 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
    10441 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
    10501 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
    10561 tgcaagctta atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag
    10621 ttagcaacat gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag
    10681 gtggtacgat cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac
    10741 cactgaattg ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg
    10801 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
    10861 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
    10921 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
    10981 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
    11041 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
    11101 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
    11161 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
    11221 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
    11281 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
    11341 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
    11401 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
    11461 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
    11521 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
    11581 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
    11641 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
    11701 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
    11761 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
    11821 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
    11881 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
    11941 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
    12001 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
    12061 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
    12121 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
    12181 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
    12241 attatcgttt cagacccacc tcccaacccc gaggggaccc ttgcgccttt tccaaggcag
    12301 ccctgggttt gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc
    12361 cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc
    12421 tacccttgtg ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt
    12481 gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc
    12541 agacggacag cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag
    12601 cggctgctca gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg
    12661 gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc
    12721 tccggagcgc acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca
    12781 gggggtaccg ccaccatgat tgaacaagat agattgcacg caggttctcc ggccgcttgg
    12841 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
    12901 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
    12961 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
    13021 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
    13081 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
    13141 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
    13201 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
    13261 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
    13321 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
    13381 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
    13441 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
    13501 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
    13561 ttctatcgcc ttcttgacga gttcttctga ggtggcgaat tctagatctt gagacaaatg
    13621 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
    13681 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
    13741 aaaattcaaa attttcgggt ttattacagg gacagcagag atccactttg gcgccggctc
    13801 gagggggccc gggtgcaaag atggataaag ttttaaacag agaggaatct ttgcagctaa
    13861 tggaccttct aggtcttgaa aggagtggga att
//
