LOCUS       sequence-171325-       13168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To produce lentiviral vector for Cas9 editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13168)
  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 13168)
  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..13168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    678..805
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1298..1531
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1715..1759
                     /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    2026..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2161..2660
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2196..2219)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2593..2612
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    2710..2734
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2740..6843
                     /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             2740..6843
                     /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             2740..6843
                     /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             6856..6876
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6883..6924
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(6974..6998)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(7021..7037)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7022..7038)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     misc_feature    7051..7639
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7104..7124)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7522..7533)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7728..7961
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(7986..8004)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7987..8004)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(8294..8313)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(8528..8547)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8647..8669
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8707..8725)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8793..8897
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8898..8966,8967..9758)
                     /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(9116..9135)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9929..10517
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10418..10437
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10671..10688
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(10758..10778)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10763..11092
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10943..11078
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          12226..12291
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             12421..12441
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    12866..13000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12903..12922)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12957..12976
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacctgaaag
      661 cgaaagggaa accagagctc tctcgacgca ggactcggct tgctgaagcg cgcgcacggc
      721 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
      781 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
      841 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
      901 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
      961 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     1021 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     1081 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     1141 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
     1201 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
     1261 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
     1321 tgggagcagc aggaagcact atgggcgcag cctcaatgac gctgacggta caggccagac
     1381 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
     1441 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
     1501 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
     1561 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattg
     1621 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     1681 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     1741 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     1801 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     1861 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     1921 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     1981 cagatccatt cgattagtga acggatctcg acggtatcgg ttaactttta aaagaaaagg
     2041 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2101 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttatcgatg gtcgagtacc
     2161 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
     2221 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
     2281 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
     2341 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
     2401 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
     2461 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
     2521 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct
     2581 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt
     2641 cctcatctcc gggcctttcg acctctagga tcctagaact agtggatccc ccgggctgca
     2701 ggaattatca caagtttgta caaaaaagca ggctccacca tggacaagaa gtactccatt
     2761 gggctcgata tcggcacaaa cagcgtcggc tgggccgtca ttacggacga gtacaaggtg
     2821 ccgagcaaaa aattcaaagt tctgggcaat accgatcgcc acagcataaa gaagaacctc
     2881 attggcgccc tcctgttcga ctccggggag acggccgaag ccacgcggct caaaagaaca
     2941 gcacggcgca gatatacccg cagaaagaat cggatctgct acctgcagga gatctttagt
     3001 aatgagatgg ctaaggtgga tgactctttc ttccataggc tggaggagtc ctttttggtg
     3061 gaggaggata aaaagcacga gcgccaccca atctttggca atatcgtgga cgaggtggcg
     3121 taccatgaaa agtacccaac catatatcat ctgaggaaga agcttgtaga cagtactgat
     3181 aaggctgact tgcggttgat ctatctcgcg ctggcgcata tgatcaaatt tcggggacac
     3241 ttcctcatcg agggggacct gaacccagac aacagcgatg tcgacaaact ctttatccaa
     3301 ctggttcaga cttacaatca gcttttcgaa gagaacccga tcaacgcatc cggagttgac
     3361 gccaaagcaa tcctgagcgc taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca
     3421 cagctccctg gggagaagaa gaacggcctg tttggtaatc ttatcgccct gtcactcggg
     3481 ctgaccccca actttaaatc taacttcgac ctggccgaag atgccaagct tcaactgagc
     3541 aaagacacct acgatgatga tctcgacaat ctgctggccc agatcggcga ccagtacgca
     3601 gacctttttt tggcggcaaa gaacctgtca gacgccattc tgctgagtga tattctgcga
     3661 gtgaacacgg agatcaccaa agctccgctg agcgctagta tgatcaagcg ctatgatgag
     3721 caccaccaag acttgacttt gctgaaggcc cttgtcagac agcaactgcc tgagaagtac
     3781 aaggaaattt tcttcgatca gtctaaaaat ggctacgccg gatacattga cggcggagca
     3841 agccaggagg aattttacaa atttattaag cccatcttgg aaaaaatgga cggcaccgag
     3901 gagctgctgg taaagcttaa cagagaagat ctgttgcgca aacagcgcac tttcgacaat
     3961 ggaagcatcc cccaccagat tcacctgggc gaactgcacg ctatcctcag gcggcaagag
     4021 gatttctacc cctttttgaa agataacagg gaaaagattg agaaaatcct cacatttcgg
     4081 ataccctact atgtaggccc cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc
     4141 aaatcagaag agaccatcac tccctggaac ttcgaggaag tcgtggataa gggggcctct
     4201 gcccagtcct tcatcgaaag gatgactaac tttgataaaa atctgcctaa cgaaaaggtg
     4261 cttcctaaac actctctgct gtacgagtac ttcacagttt ataacgagct caccaaggtc
     4321 aaatacgtca cagaagggat gagaaagcca gcattcctgt ctggagagca gaagaaagct
     4381 atcgtggacc tcctcttcaa gacgaaccgg aaagttaccg tgaaacagct caaagaagac
     4441 tatttcaaaa agattgaatg tttcgactct gttgaaatca gcggagtgga ggatcgcttc
     4501 aacgcatccc tgggaacgta tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg
     4561 gacaatgagg agaacgagga cattcttgag gacattgtcc tcacccttac gttgtttgaa
     4621 gatagggaga tgattgaaga acgcttgaaa acttacgctc atctcttcga cgacaaagtc
     4681 atgaaacagc tcaagaggcg ccgatataca ggatgggggc ggctgtcaag aaaactgatc
     4741 aatgggatcc gagacaagca gagtggaaag acaatcctgg attttcttaa gtccgatgga
     4801 tttgccaacc ggaacttcat gcagttgatc catgatgact ctctcacctt taaggaggac
     4861 atccagaaag cacaagtttc tggccagggg gacagtcttc acgagcacat cgctaatctt
     4921 gcaggtagcc cagctatcaa aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc
     4981 gtcaaagtaa tgggaaggca taagcccgag aatatcgtta tcgagatggc ccgagagaac
     5041 caaactaccc agaagggaca gaagaacagt agggaaagga tgaagaggat tgaagagggt
     5101 ataaaagaac tggggtccca aatccttaag gaacacccag ttgaaaacac ccagcttcag
     5161 aatgagaagc tctacctgta ctacctgcag aacggcaggg acatgtacgt ggatcaggaa
     5221 ctggacatca atcggctctc cgactacgac gtggatcata tcgtgcccca gtcttttctc
     5281 aaagatgatt ctattgataa taaagtgttg acaagatccg ataaaaatag agggaagagt
     5341 gataacgtcc cctcagaaga agttgtcaag aaaatgaaaa attattggcg gcagctgctg
     5401 aacgccaaac tgatcacaca acggaagttc gataatctga ctaaggctga acgaggtggc
     5461 ctgtctgagt tggataaagc cggcttcatc aaaaggcagc ttgttgagac acgccagatc
     5521 accaagcacg tggcccaaat tctcgattca cgcatgaaca ccaagtacga tgaaaatgac
     5581 aaactgattc gagaggtgaa agttattact ctgaagtcta agctggtctc agatttcaga
     5641 aaggactttc agttttataa ggtgagagag atcaacaatt accaccatgc gcatgatgcc
     5701 tacctgaatg cagtggtagg cactgcactt atcaaaaaat atcccaagct tgaatctgaa
     5761 tttgtttacg gagactataa agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag
     5821 gaaataggca aggccaccgc taagtacttc ttttacagca atattatgaa ttttttcaag
     5881 accgagatta cactggccaa tggagagatt cggaagcgac cacttatcga aacaaacgga
     5941 gaaacaggag aaatcgtgtg ggacaagggt agggatttcg cgacagtccg gaaggtcctg
     6001 tccatgccgc aggtgaacat cgttaaaaag accgaagtac agaccggagg cttctccaag
     6061 gaaagtatcc tcccgaaaag gaacagcgac aagctgatcg cacgcaaaaa agattgggac
     6121 cccaagaaat acggcggatt cgattctcct acagtcgctt acagtgtact ggttgtggcc
     6181 aaagtggaga aagggaagtc taaaaaactc aaaagcgtca aggaactgct gggcatcaca
     6241 atcatggagc gatcaagctt cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat
     6301 aaagaggtca aaaaagacct catcattaag cttcccaagt actctctctt tgagcttgaa
     6361 aacggccgga aacgaatgct cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca
     6421 ctgccctcta aatacgttaa tttcttgtat ctggccagcc actatgaaaa gctcaaaggg
     6481 tctcccgaag ataatgagca gaagcagctg ttcgtggaac aacacaaaca ctaccttgat
     6541 gagatcatcg agcaaataag cgaattctcc aaaagagtga tcctcgccga cgctaacctc
     6601 gataaggtgc tttctgctta caataagcac agggataagc ccatcaggga gcaggcagaa
     6661 aacattatcc acttgtttac tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc
     6721 gacaccacca tagacagaaa gcggtacacc tctacaaagg aggtcctgga cgccacactg
     6781 attcatcagt caattacggg gctctatgaa acaagaatcg acctctctca gctcggtgga
     6841 gacagcaggg ctgaccccaa gaagaagagg aaggtgggta ccggtaagcc tatccctaac
     6901 cctctcctcg gtctcgattc tacgtgagaa ttcaagggtt cgatccctac cggttagtaa
     6961 tgagtttatc tagacccagc tttcttgtac aaagtggtga taattcgata tcaagcttat
     7021 cgataccgtc gacctcgagg gaattccgat aatcaacctc tggattacaa aatttgtgaa
     7081 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     7141 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     7201 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     7261 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     7321 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     7381 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     7441 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     7501 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     7561 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     7621 ctttgggccg cctccccgca tcgggaattc gagctcggta cctttaagac caatgactta
     7681 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     7741 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
     7801 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     7861 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     7921 atccctcaga cccttttagt cagtgtggaa aatctctagc agcatctaga attaattccg
     7981 tgtattctat agtgtcacct aaatcgtatg tgtatgatac ataaggttat gtattaattg
     8041 tagccgcgtt ctaacgacaa tatgtacaag cctaattgtg tagcatctgg cttactgaag
     8101 cagaccctat catctctctc gtaaactgcc gtcagagtcg gtttggttgg acgaaccttc
     8161 tgagtttctg gtaacgccgt cccgcacccg gaaatggtca gcgaaccaat cagcagggtc
     8221 atcgctagcc agatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
     8281 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
     8341 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
     8401 ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
     8461 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg atatggtgca
     8521 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     8581 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     8641 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     8701 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     8761 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     8821 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     8881 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     8941 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     9001 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     9061 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     9121 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     9181 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     9241 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     9301 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     9361 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     9421 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     9481 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     9541 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     9601 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     9661 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     9721 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     9781 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     9841 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     9901 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     9961 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
    10021 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
    10081 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
    10141 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
    10201 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
    10261 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
    10321 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
    10381 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
    10441 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
    10501 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
    10561 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
    10621 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
    10681 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
    10741 ttcattaatg cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
    10801 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
    10861 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
    10921 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
    10981 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
    11041 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttgg
    11101 acacaagaca ggcttgcgag atatgtttga gaataccact ttatcccgcg tcagggagag
    11161 gcagtgcgta aaaagacgcg gactcatgtg aaatactggt ttttagtgcg ccagatctct
    11221 ataatctcgc gcaacctatt ttcccctcga acacttttta agccgtagat aaacaggctg
    11281 ggacacttca catgagcgaa aaatacatcg tcacctggga catgttgcag atccatgcac
    11341 gtaaactcgc aagccgactg atgccttctg aacaatggaa aggcattatt gccgtaagcc
    11401 gtggcggtct gtaccgggtg cgttactggc gcgtgaactg ggtattcgtc atgtcgatac
    11461 cgtttgtatt tccagctacg atcacgacaa ccagcgcgag cttaaagtgc tgaaacgcgc
    11521 agaaggcgat ggcgaaggct tcatcgttat tgatgacctg gtggataccg gtggtactgc
    11581 ggttgcgatt cgtgaaatgt atccaaaagc gcactttgtc accatcttcg caaaaccggc
    11641 tggtcgtccg ctggttgatg actatgttgt tgatatcccg caagatacct ggattgaaca
    11701 gccgtgggat atgggcgtcg tattcgtccc gccaatctcc ggtcgctaat cttttcaacg
    11761 cctggcactg ccgggcgttg ttctttttaa cttcaggcgg gttacaatag tttccagtaa
    11821 gtattctgga ggctgcatcc atgacacagg caaacctgag cgaaaccctg ttcaaacccc
    11881 gctttaaaca tcctgaaacc tcgacgctag tccgccgctt taatcacggc gcacaaccgc
    11941 ctgtgcagtc ggcccttgat ggtaaaacca tccctcactg gtatcgcatg attaaccgtc
    12001 tgatgtggat ctggcgcggc attgacccac gcgaaatcct cgacgtccag gcacgtattg
    12061 tgatgagcga tgccgaacgt accgacgatg atttatacga tacggtgatt ggctaccgtg
    12121 gcggcaactg gatttatgag tgggccccgg atctttgtga aggaacctta cttctgtggt
    12181 gtgacataat tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt
    12241 ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta gattccaacc
    12301 tatggaactg atgaatggga gcagtggtgg aatgccttta atgaggaaaa cctgttttgc
    12361 tcagaagaaa tgccatctag tgatgatgag gctactgctg actctcaaca ttctactcct
    12421 ccaaaaaaga agagaaaggt agaagacccc aaggactttc cttcagaatt gctaagtttt
    12481 ttgagtcatg ctgtgtttag taatagaact cttgcttgct ttgctattta caccacaaag
    12541 gaaaaagctg cactgctata caagaaaatt atggaaaaat attctgtaac ctttataagt
    12601 aggcataaca gttataatca taacatactg ttttttctta ctccacacag gcatagagtg
    12661 tctgctatta ataactatgc tcaaaaattg tgtaccttta gctttttaat ttgtaaaggg
    12721 gttaataagg aatatttgat gtatagtgcc ttgactagag atcataatca gccataccac
    12781 atttgtagag cttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
    12841 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
    12901 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
    12961 tttgtccaaa ctcatcaatg tatcttatca tgtgtggatc aactggataa ctcaagctaa
    13021 ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaaat tacctgtggt
    13081 ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt
    13141 gttaaatatg tactacaaac ttagtagt
//
