LOCUS       sequence-175351-       13016 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  To produce lentiviral vector for Cas9 editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 13016)
  AUTHORS   Merienne N, Vachey G, de Longprez L, Meunier C, Zimmer V, Perriard 
            G, Canales M, Mathias A, Herrgott L, Beltraminelli T, Maulet A, 
            Dequesne T, Pythoud C, Rey M, Pellerin L, Brouillet E, Perrier AL, 
            du Pasquier R, Deglon N
  TITLE     The Self-Inactivating KamiCas9 System for the Editing of CNS Disease
            Genes.
  JOURNAL   Cell Rep. 2017 Sep 19;20(12):2980-2991. doi: 
            10.1016/j.celrep.2017.08.075.
  PUBMED    28930690
REFERENCE   2  (bases 1 to 13016)
  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"; doi: 
            "10.1016/j.celrep.2017.08"; journalName: "Cell Rep"; date: 
            "2017-09-19- 19"; volume: "20"; issue: "12"; pages: "2980-2991"
COMMENT     Sequence Label: SIN-PGK-Cas9-V5-WPRE
FEATURES             Location/Qualifiers
     source          1..13016
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    37..61
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             67..4170
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             67..4170
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             67..4170
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4183..4203
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4210..4251
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(4301..4325)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4348..4364)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4349..4365)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     misc_feature    4378..4966
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4431..4451)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4849..4860)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5055..5288
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(5313..5331)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5314..5331)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5621..5640)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5854..5873)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5973..5995
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6033..6051)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6119..6223
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6224..6292,6293..7084)
                     /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(6442..6461)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7255..7843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7744..7763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7997..8014
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(8084..8104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8089..8418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8269..8404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          9552..9617
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             9747..9767
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    10192..10326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10229..10248)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10283..10302
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10494..11127
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    11174..11299
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11792..12025
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12210..12254
                     /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    12521..12638
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     complement(12691..12714)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 tagaactagt ggatcccccg ggctgcagga attatcacaa gtttgtacaa aaaagcaggc
       61 tccaccatgg acaagaagta ctccattggg ctcgatatcg gcacaaacag cgtcggctgg
      121 gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc
      181 gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggagacg
      241 gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg
      301 atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc
      361 cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc
      421 tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg
      481 aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg
      541 gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac
      601 agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag
      661 aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa
      721 tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa cggcctgttt
      781 ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa cttcgacctg
      841 gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct cgacaatctg
      901 ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa cctgtcagac
      961 gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc tccgctgagc
     1021 gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct gaaggccctt
     1081 gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc taaaaatggc
     1141 tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt tattaagccc
     1201 atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag agaagatctg
     1261 ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca cctgggcgaa
     1321 ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga taacagggaa
     1381 aagattgaga aaatcctcac atttcggata ccctactatg taggccccct cgcccgggga
     1441 aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc ctggaacttc
     1501 gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat gactaacttt
     1561 gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta cgagtacttc
     1621 acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag aaagccagca
     1681 ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac gaaccggaaa
     1741 gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt cgactctgtt
     1801 gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca cgatctcctg
     1861 aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat tcttgaggac
     1921 attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg cttgaaaact
     1981 tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg atatacagga
     2041 tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag tggaaagaca
     2101 atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca gttgatccat
     2161 gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg ccagggggac
     2221 agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa gggaatactg
     2281 cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat
     2341 atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa gaacagtagg
     2401 gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat ccttaaggaa
     2461 cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta cctgcagaac
     2521 ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga ctacgacgtg
     2581 gatcatatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa agtgttgaca
     2641 agatccgata aaaatagagg gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa
     2701 atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg gaagttcgat
     2761 aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagccgg cttcatcaaa
     2821 aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct cgattcacgc
     2881 atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt tattactctg
     2941 aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt gagagagatc
     3001 aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac tgcacttatc
     3061 aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt gtacgatgtt
     3121 aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt
     3181 tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg agagattcgg
     3241 aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga caagggtagg
     3301 gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc
     3361 gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag
     3421 ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga ttctcctaca
     3481 gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa
     3541 agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga aaaaaacccc
     3601 atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat cattaagctt
     3661 cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc
     3721 gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt cttgtatctg
     3781 gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa gcagctgttc
     3841 gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga attctccaaa
     3901 agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa taagcacagg
     3961 gataagccca tcagggagca ggcagaaaac attatccact tgtttactct gaccaacttg
     4021 ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg gtacacctct
     4081 acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct ctatgaaaca
     4141 agaatcgacc tctctcagct cggtggagac agcagggctg accccaagaa gaagaggaag
     4201 gtgggtaccg gtaagcctat ccctaaccct ctcctcggtc tcgattctac gtgagaattc
     4261 aagggttcga tccctaccgg ttagtaatga gtttatctag acccagcttt cttgtacaaa
     4321 gtggtgataa ttcgatatca agcttatcga taccgtcgac ctcgagggaa ttccgataat
     4381 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     4441 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     4501 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     4561 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     4621 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     4681 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     4741 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     4801 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     4861 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     4921 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ggaattcgag
     4981 ctcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     5041 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     5101 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     5161 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     5221 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     5281 ctctagcagc atctagaatt aattccgtgt attctatagt gtcacctaaa tcgtatgtgt
     5341 atgatacata aggttatgta ttaattgtag ccgcgttcta acgacaatat gtacaagcct
     5401 aattgtgtag catctggctt actgaagcag accctatcat ctctctcgta aactgccgtc
     5461 agagtcggtt tggttggacg aaccttctga gtttctggta acgccgtccc gcacccggaa
     5521 atggtcagcg aaccaatcag cagggtcatc gctagccaga tcctctacgc cggacgcatc
     5581 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
     5641 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
     5701 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt
     5761 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
     5821 cataagggag agcgtcgaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     5881 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     5941 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     6001 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     6061 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     6121 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6181 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6241 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6301 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6361 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6421 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     6481 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6541 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6601 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6661 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6721 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6781 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6841 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6901 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6961 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7021 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7081 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7141 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7201 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7261 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7321 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7381 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     7441 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7501 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7561 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7621 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7681 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7741 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7801 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7861 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7921 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     7981 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     8041 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tgtggaatgt gtgtcagtta
     8101 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     8161 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     8221 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     8281 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     8341 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     8401 ggcctaggct tttgcaaaaa gcttggacac aagacaggct tgcgagatat gtttgagaat
     8461 accactttat cccgcgtcag ggagaggcag tgcgtaaaaa gacgcggact catgtgaaat
     8521 actggttttt agtgcgccag atctctataa tctcgcgcaa cctattttcc cctcgaacac
     8581 tttttaagcc gtagataaac aggctgggac acttcacatg agcgaaaaat acatcgtcac
     8641 ctgggacatg ttgcagatcc atgcacgtaa actcgcaagc cgactgatgc cttctgaaca
     8701 atggaaaggc attattgccg taagccgtgg cggtctgtac cgggtgcgtt actggcgcgt
     8761 gaactgggta ttcgtcatgt cgataccgtt tgtatttcca gctacgatca cgacaaccag
     8821 cgcgagctta aagtgctgaa acgcgcagaa ggcgatggcg aaggcttcat cgttattgat
     8881 gacctggtgg ataccggtgg tactgcggtt gcgattcgtg aaatgtatcc aaaagcgcac
     8941 tttgtcacca tcttcgcaaa accggctggt cgtccgctgg ttgatgacta tgttgttgat
     9001 atcccgcaag atacctggat tgaacagccg tgggatatgg gcgtcgtatt cgtcccgcca
     9061 atctccggtc gctaatcttt tcaacgcctg gcactgccgg gcgttgttct ttttaacttc
     9121 aggcgggtta caatagtttc cagtaagtat tctggaggct gcatccatga cacaggcaaa
     9181 cctgagcgaa accctgttca aaccccgctt taaacatcct gaaacctcga cgctagtccg
     9241 ccgctttaat cacggcgcac aaccgcctgt gcagtcggcc cttgatggta aaaccatccc
     9301 tcactggtat cgcatgatta accgtctgat gtggatctgg cgcggcattg acccacgcga
     9361 aatcctcgac gtccaggcac gtattgtgat gagcgatgcc gaacgtaccg acgatgattt
     9421 atacgatacg gtgattggct accgtggcgg caactggatt tatgagtggg ccccggatct
     9481 ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct acagagattt
     9541 aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa
     9601 ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag tggtggaatg
     9661 cctttaatga ggaaaacctg ttttgctcag aagaaatgcc atctagtgat gatgaggcta
     9721 ctgctgactc tcaacattct actcctccaa aaaagaagag aaaggtagaa gaccccaagg
     9781 actttccttc agaattgcta agttttttga gtcatgctgt gtttagtaat agaactcttg
     9841 cttgctttgc tatttacacc acaaaggaaa aagctgcact gctatacaag aaaattatgg
     9901 aaaaatattc tgtaaccttt ataagtaggc ataacagtta taatcataac atactgtttt
     9961 ttcttactcc acacaggcat agagtgtctg ctattaataa ctatgctcaa aaattgtgta
    10021 cctttagctt tttaatttgt aaaggggtta ataaggaata tttgatgtat agtgccttga
    10081 ctagagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
    10141 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
    10201 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
    10261 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
    10321 tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac cccataccct
    10381 attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta
    10441 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agttggaagg
    10501 gctaattcac tcccaaagaa gacaagatat ccttgatctg tggatctacc acacacaagg
    10561 ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt
    10621 tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg
    10681 agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
    10741 agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca
    10801 tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga
    10861 ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat
    10921 ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
    10981 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
    11041 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
    11101 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
    11161 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
    11221 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
    11281 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
    11341 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
    11401 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
    11461 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
    11521 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
    11581 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
    11641 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
    11701 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
    11761 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
    11821 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
    11881 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
    11941 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
    12001 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
    12061 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
    12121 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
    12181 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
    12241 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
    12301 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
    12361 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
    12421 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
    12481 ccattcgatt agtgaacgga tctcgacggt atcggttaac ttttaaaaga aaagggggga
    12541 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
    12601 aagaattaca aaaacaaatt acaaaaattc aaaattttat cgatggtcga gtaccgggta
    12661 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact
    12721 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa
    12781 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag
    12841 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc
    12901 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc
    12961 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aagggg
//
