LOCUS       sequence-175362-       13258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To produce lentiviral vector for gene editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13258)
  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 13258)
  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-CMV-Cas9-V5-WPRE
FEATURES             Location/Qualifiers
     source          1..13258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      106..241
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          1389..1454
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1584..1604
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2029..2163
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2066..2085)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2120..2139
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             2331..2964
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    3011..3136
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3629..3862
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4047..4091
                     /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    4358..4475
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        4498..4801
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4802..5005
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4956..4976
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5002..5026
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    5132..5156
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5162..9265
                     /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             5162..9265
                     /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             5162..9265
                     /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             9278..9298
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9305..9346
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(9396..9420)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(9443..9459)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9444..9460)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     misc_feature    9473..10061
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9526..9546)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9944..9955)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10150..10383
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(10408..10426)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(10409..10426)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(10716..10735)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(10949..10968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11068..11090
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(11128..11146)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        11214..11318
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11319..11387,11388..12179)
                     /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(11537..11556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12350..12938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12839..12858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13092..13109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(13179..13199)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(13184..13258,1..255)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
       61 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
      121 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
      181 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
      241 ctaggctttt gcaaaaagct tggacacaag acaggcttgc gagatatgtt tgagaatacc
      301 actttatccc gcgtcaggga gaggcagtgc gtaaaaagac gcggactcat gtgaaatact
      361 ggtttttagt gcgccagatc tctataatct cgcgcaacct attttcccct cgaacacttt
      421 ttaagccgta gataaacagg ctgggacact tcacatgagc gaaaaataca tcgtcacctg
      481 ggacatgttg cagatccatg cacgtaaact cgcaagccga ctgatgcctt ctgaacaatg
      541 gaaaggcatt attgccgtaa gccgtggcgg tctgtaccgg gtgcgttact ggcgcgtgaa
      601 ctgggtattc gtcatgtcga taccgtttgt atttccagct acgatcacga caaccagcgc
      661 gagcttaaag tgctgaaacg cgcagaaggc gatggcgaag gcttcatcgt tattgatgac
      721 ctggtggata ccggtggtac tgcggttgcg attcgtgaaa tgtatccaaa agcgcacttt
      781 gtcaccatct tcgcaaaacc ggctggtcgt ccgctggttg atgactatgt tgttgatatc
      841 ccgcaagata cctggattga acagccgtgg gatatgggcg tcgtattcgt cccgccaatc
      901 tccggtcgct aatcttttca acgcctggca ctgccgggcg ttgttctttt taacttcagg
      961 cgggttacaa tagtttccag taagtattct ggaggctgca tccatgacac aggcaaacct
     1021 gagcgaaacc ctgttcaaac cccgctttaa acatcctgaa acctcgacgc tagtccgccg
     1081 ctttaatcac ggcgcacaac cgcctgtgca gtcggccctt gatggtaaaa ccatccctca
     1141 ctggtatcgc atgattaacc gtctgatgtg gatctggcgc ggcattgacc cacgcgaaat
     1201 cctcgacgtc caggcacgta ttgtgatgag cgatgccgaa cgtaccgacg atgatttata
     1261 cgatacggtg attggctacc gtggcggcaa ctggatttat gagtgggccc cggatctttg
     1321 tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa
     1381 gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg
     1441 tttgtgtatt ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcct
     1501 ttaatgagga aaacctgttt tgctcagaag aaatgccatc tagtgatgat gaggctactg
     1561 ctgactctca acattctact cctccaaaaa agaagagaaa ggtagaagac cccaaggact
     1621 ttccttcaga attgctaagt tttttgagtc atgctgtgtt tagtaataga actcttgctt
     1681 gctttgctat ttacaccaca aaggaaaaag ctgcactgct atacaagaaa attatggaaa
     1741 aatattctgt aacctttata agtaggcata acagttataa tcataacata ctgttttttc
     1801 ttactccaca caggcataga gtgtctgcta ttaataacta tgctcaaaaa ttgtgtacct
     1861 ttagcttttt aatttgtaaa ggggttaata aggaatattt gatgtatagt gccttgacta
     1921 gagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     1981 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2041 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2101 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     2161 atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt
     2221 accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc tgaattattt
     2281 tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt tggaagggct
     2341 aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
     2401 cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac tgacctttgg
     2461 atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca ataaaggaga
     2521 gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
     2581 gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag agctgcatcc
     2641 ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg ctggggactt
     2701 tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat cctgcatata
     2761 agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc
     2821 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     2881 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     2941 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
     3001 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
     3061 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
     3121 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
     3181 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
     3241 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
     3301 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
     3361 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
     3421 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc
     3481 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
     3541 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     3601 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     3661 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     3721 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     3781 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
     3841 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     3901 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
     3961 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     4021 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     4081 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
     4141 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
     4201 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
     4261 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
     4321 ttcgattagt gaacggatct cgacggtatc ggttaacttt taaaagaaaa ggggggattg
     4381 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     4441 aattacaaaa acaaattaca aaaattcaaa attttatcga taagcttggg agttccgcgt
     4501 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     4561 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     4621 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     4681 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     4741 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     4801 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     4861 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     4921 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     4981 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     5041 tccacgctgt tttgacctcc atagaagaca ccgactctag ctagaggatc ttccctagaa
     5101 ctagtggatc ccccgggctg caggaattat cacaagtttg tacaaaaaag caggctccac
     5161 catggacaag aagtactcca ttgggctcga tatcggcaca aacagcgtcg gctgggccgt
     5221 cattacggac gagtacaagg tgccgagcaa aaaattcaaa gttctgggca ataccgatcg
     5281 ccacagcata aagaagaacc tcattggcgc cctcctgttc gactccgggg agacggccga
     5341 agccacgcgg ctcaaaagaa cagcacggcg cagatatacc cgcagaaaga atcggatctg
     5401 ctacctgcag gagatcttta gtaatgagat ggctaaggtg gatgactctt tcttccatag
     5461 gctggaggag tcctttttgg tggaggagga taaaaagcac gagcgccacc caatctttgg
     5521 caatatcgtg gacgaggtgg cgtaccatga aaagtaccca accatatatc atctgaggaa
     5581 gaagcttgta gacagtactg ataaggctga cttgcggttg atctatctcg cgctggcgca
     5641 tatgatcaaa tttcggggac acttcctcat cgagggggac ctgaacccag acaacagcga
     5701 tgtcgacaaa ctctttatcc aactggttca gacttacaat cagcttttcg aagagaaccc
     5761 gatcaacgca tccggagttg acgccaaagc aatcctgagc gctaggctgt ccaaatcccg
     5821 gcggctcgaa aacctcatcg cacagctccc tggggagaag aagaacggcc tgtttggtaa
     5881 tcttatcgcc ctgtcactcg ggctgacccc caactttaaa tctaacttcg acctggccga
     5941 agatgccaag cttcaactga gcaaagacac ctacgatgat gatctcgaca atctgctggc
     6001 ccagatcggc gaccagtacg cagacctttt tttggcggca aagaacctgt cagacgccat
     6061 tctgctgagt gatattctgc gagtgaacac ggagatcacc aaagctccgc tgagcgctag
     6121 tatgatcaag cgctatgatg agcaccacca agacttgact ttgctgaagg cccttgtcag
     6181 acagcaactg cctgagaagt acaaggaaat tttcttcgat cagtctaaaa atggctacgc
     6241 cggatacatt gacggcggag caagccagga ggaattttac aaatttatta agcccatctt
     6301 ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt aacagagaag atctgttgcg
     6361 caaacagcgc actttcgaca atggaagcat cccccaccag attcacctgg gcgaactgca
     6421 cgctatcctc aggcggcaag aggatttcta cccctttttg aaagataaca gggaaaagat
     6481 tgagaaaatc ctcacatttc ggatacccta ctatgtaggc cccctcgccc ggggaaattc
     6541 cagattcgcg tggatgactc gcaaatcaga agagaccatc actccctgga acttcgagga
     6601 agtcgtggat aagggggcct ctgcccagtc cttcatcgaa aggatgacta actttgataa
     6661 aaatctgcct aacgaaaagg tgcttcctaa acactctctg ctgtacgagt acttcacagt
     6721 ttataacgag ctcaccaagg tcaaatacgt cacagaaggg atgagaaagc cagcattcct
     6781 gtctggagag cagaagaaag ctatcgtgga cctcctcttc aagacgaacc ggaaagttac
     6841 cgtgaaacag ctcaaagaag actatttcaa aaagattgaa tgtttcgact ctgttgaaat
     6901 cagcggagtg gaggatcgct tcaacgcatc cctgggaacg tatcacgatc tcctgaaaat
     6961 cattaaagac aaggacttcc tggacaatga ggagaacgag gacattcttg aggacattgt
     7021 cctcaccctt acgttgtttg aagataggga gatgattgaa gaacgcttga aaacttacgc
     7081 tcatctcttc gacgacaaag tcatgaaaca gctcaagagg cgccgatata caggatgggg
     7141 gcggctgtca agaaaactga tcaatgggat ccgagacaag cagagtggaa agacaatcct
     7201 ggattttctt aagtccgatg gatttgccaa ccggaacttc atgcagttga tccatgatga
     7261 ctctctcacc tttaaggagg acatccagaa agcacaagtt tctggccagg gggacagtct
     7321 tcacgagcac atcgctaatc ttgcaggtag cccagctatc aaaaagggaa tactgcagac
     7381 cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg cataagcccg agaatatcgt
     7441 tatcgagatg gcccgagaga accaaactac ccagaaggga cagaagaaca gtagggaaag
     7501 gatgaagagg attgaagagg gtataaaaga actggggtcc caaatcctta aggaacaccc
     7561 agttgaaaac acccagcttc agaatgagaa gctctacctg tactacctgc agaacggcag
     7621 ggacatgtac gtggatcagg aactggacat caatcggctc tccgactacg acgtggatca
     7681 tatcgtgccc cagtcttttc tcaaagatga ttctattgat aataaagtgt tgacaagatc
     7741 cgataaaaat agagggaaga gtgataacgt cccctcagaa gaagttgtca agaaaatgaa
     7801 aaattattgg cggcagctgc tgaacgccaa actgatcaca caacggaagt tcgataatct
     7861 gactaaggct gaacgaggtg gcctgtctga gttggataaa gccggcttca tcaaaaggca
     7921 gcttgttgag acacgccaga tcaccaagca cgtggcccaa attctcgatt cacgcatgaa
     7981 caccaagtac gatgaaaatg acaaactgat tcgagaggtg aaagttatta ctctgaagtc
     8041 taagctggtc tcagatttca gaaaggactt tcagttttat aaggtgagag agatcaacaa
     8101 ttaccaccat gcgcatgatg cctacctgaa tgcagtggta ggcactgcac ttatcaaaaa
     8161 atatcccaag cttgaatctg aatttgttta cggagactat aaagtgtacg atgttaggaa
     8221 aatgatcgca aagtctgagc aggaaatagg caaggccacc gctaagtact tcttttacag
     8281 caatattatg aattttttca agaccgagat tacactggcc aatggagaga ttcggaagcg
     8341 accacttatc gaaacaaacg gagaaacagg agaaatcgtg tgggacaagg gtagggattt
     8401 cgcgacagtc cggaaggtcc tgtccatgcc gcaggtgaac atcgttaaaa agaccgaagt
     8461 acagaccgga ggcttctcca aggaaagtat cctcccgaaa aggaacagcg acaagctgat
     8521 cgcacgcaaa aaagattggg accccaagaa atacggcgga ttcgattctc ctacagtcgc
     8581 ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag tctaaaaaac tcaaaagcgt
     8641 caaggaactg ctgggcatca caatcatgga gcgatcaagc ttcgaaaaaa accccatcga
     8701 ctttctcgag gcgaaaggat ataaagaggt caaaaaagac ctcatcatta agcttcccaa
     8761 gtactctctc tttgagcttg aaaacggccg gaaacgaatg ctcgctagtg cgggcgagct
     8821 gcagaaaggt aacgagctgg cactgccctc taaatacgtt aatttcttgt atctggccag
     8881 ccactatgaa aagctcaaag ggtctcccga agataatgag cagaagcagc tgttcgtgga
     8941 acaacacaaa cactaccttg atgagatcat cgagcaaata agcgaattct ccaaaagagt
     9001 gatcctcgcc gacgctaacc tcgataaggt gctttctgct tacaataagc acagggataa
     9061 gcccatcagg gagcaggcag aaaacattat ccacttgttt actctgacca acttgggcgc
     9121 gcctgcagcc ttcaagtact tcgacaccac catagacaga aagcggtaca cctctacaaa
     9181 ggaggtcctg gacgccacac tgattcatca gtcaattacg gggctctatg aaacaagaat
     9241 cgacctctct cagctcggtg gagacagcag ggctgacccc aagaagaaga ggaaggtggg
     9301 taccggtaag cctatcccta accctctcct cggtctcgat tctacgtgag aattcaaggg
     9361 ttcgatccct accggttagt aatgagttta tctagaccca gctttcttgt acaaagtggt
     9421 gataattcga tatcaagctt atcgataccg tcgacctcga gggaattccg ataatcaacc
     9481 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     9541 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     9601 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     9661 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     9721 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     9781 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     9841 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     9901 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     9961 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
    10021 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgggaat tcgagctcgg
    10081 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
    10141 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
    10201 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
    10261 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
    10321 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
    10381 gcagcatcta gaattaattc cgtgtattct atagtgtcac ctaaatcgta tgtgtatgat
    10441 acataaggtt atgtattaat tgtagccgcg ttctaacgac aatatgtaca agcctaattg
    10501 tgtagcatct ggcttactga agcagaccct atcatctctc tcgtaaactg ccgtcagagt
    10561 cggtttggtt ggacgaacct tctgagtttc tggtaacgcc gtcccgcacc cggaaatggt
    10621 cagcgaacca atcagcaggg tcatcgctag ccagatcctc tacgccggac gcatcgtggc
    10681 cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg
    10741 ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc
    10801 aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc
    10861 ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa
    10921 gggagagcgt cgaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
    10981 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
    11041 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
    11101 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
    11161 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
    11221 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
    11281 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
    11341 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
    11401 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
    11461 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
    11521 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
    11581 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
    11641 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
    11701 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
    11761 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
    11821 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
    11881 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
    11941 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
    12001 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
    12061 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
    12121 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
    12181 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
    12241 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
    12301 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
    12361 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
    12421 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
    12481 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
    12541 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
    12601 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
    12661 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
    12721 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
    12781 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
    12841 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
    12901 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
    12961 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
    13021 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
    13081 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
    13141 ctctccccgc gcgttggccg attcattaat gcagctgtgg aatgtgtgtc agttagggtg
    13201 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattag
//
