LOCUS       sequence-391662-        9446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression construct encoding SpCas9 nuclease and an AAVS1-targeting
            guide RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9446)
  AUTHORS   Wang Q, Liu J, Janssen JM, Le Bouteiller M, Frock RL, Goncalves MAFV
  TITLE     Precise and broad scope genome editing based on high-specificity 
            Cas9 nickases.
  JOURNAL   Nucleic Acids Res. 2021 Jan 25;49(2):1173-1198. doi: 
            10.1093/nar/gkaa1236.
  PUBMED    33398349
REFERENCE   2  (bases 1 to 9446)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2021-01-25- 25"; volume: "49"; issue: 
            "2"; pages: "1173-1198"
COMMENT     Sequence Label: AV15_pCAG.Cas9.gRNA.S1
FEATURES             Location/Qualifiers
     source          1..9446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        212..460
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     212..232
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     383..402
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        481..556
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        758..1137
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1139..1414
                     /label=chicken beta-actin promoter
     primer_bind     2344..2366
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2428..2447
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2518..2527
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2524..6627
                     /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             2524..6627
                     /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             2524..6627
                     /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             6640..6660
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6715..6734)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6759..6807
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        complement(7253..7271)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7326..7344)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7412..7516
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7517..7585,7586..8377)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7735..7754)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8548..9136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9037..9056
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9290..9307
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     9374..9390
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9397..9416
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9397..9415
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 atgttattcc tccttattta aaagcttaag gtgcacggcc cacgtggcca ctagtacttc
       61 tcgagctctg tacatgtccg cggtcgcgac gtacgcgtat cgatggcgcc agctgcaggc
      121 ggccgccata tgcatcctag gcctattaat attccggagt atacgtagcc ggctaacgtt
      181 gactggatcc ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata
      241 tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac
      301 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
      361 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
      421 tttcttggct ttatatatct tgtggaaagg acgaaacacc ggggccacta gggacaggat
      481 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
      541 ggcaccgagt cggtgctttt tttctagacc cagctttctt gtacaacaac cggtacctct
      601 agaactatag ctagcatgcg caaatttaaa gcgctgatat cgatcgcgcg cagatctgtc
      661 atgatgatca ttgcaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
      721 gcgatgtacg ggccagatat actcgagcca tggattcgac attgattatt gactagttat
      781 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      841 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      901 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      961 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     1021 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     1081 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
     1141 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
     1201 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg ggggggggsg
     1261 cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc
     1321 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
     1381 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc
     1441 cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact
     1501 cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta
     1561 atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc
     1621 tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc
     1681 gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc
     1741 gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggg
     1801 ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt
     1861 gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg ctgagcacgg
     1921 cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg tggtggcggc
     1981 aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc
     2041 gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt
     2101 tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga
     2161 aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg
     2221 gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc
     2281 ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg
     2341 cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg
     2401 ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
     2461 ttggcaaaga attatcgcat gcctgcagag ctctagagtc tagaggatcg aacccttgcc
     2521 accatggaca agaagtactc cattgggctc gatatcggca caaacagcgt cggctgggcc
     2581 gtcattacgg acgagtacaa ggtgccgagc aaaaaattca aagttctggg caataccgat
     2641 cgccacagca taaagaagaa cctcattggc gccctcctgt tcgactccgg ggagacggcc
     2701 gaagccacgc ggctcaaaag aacagcacgg cgcagatata cccgcagaaa gaatcggatc
     2761 tgctacctgc aggagatctt tagtaatgag atggctaagg tggatgactc tttcttccat
     2821 aggctggagg agtccttttt ggtggaggag gataaaaagc acgagcgcca cccaatcttt
     2881 ggcaatatcg tggacgaggt ggcgtaccat gaaaagtacc caaccatata tcatctgagg
     2941 aagaagcttg tagacagtac tgataaggct gacttgcggt tgatctatct cgcgctggcg
     3001 catatgatca aatttcgggg acacttcctc atcgaggggg acctgaaccc agacaacagc
     3061 gatgtcgaca aactctttat ccaactggtt cagacttaca atcagctttt cgaagagaac
     3121 ccgatcaacg catccggagt tgacgccaaa gcaatcctga gcgctaggct gtccaaatcc
     3181 cggcggctcg aaaacctcat cgcacagctc cctggggaga agaagaacgg cctgtttggt
     3241 aatcttatcg ccctgtcact cgggctgacc cccaacttta aatctaactt cgacctggcc
     3301 gaagatgcca agcttcaact gagcaaagac acctacgatg atgatctcga caatctgctg
     3361 gcccagatcg gcgaccagta cgcagacctt tttttggcgg caaagaacct gtcagacgcc
     3421 attctgctga gtgatattct gcgagtgaac acggagatca ccaaagctcc gctgagcgct
     3481 agtatgatca agcgctatga tgagcaccac caagacttga ctttgctgaa ggcccttgtc
     3541 agacagcaac tgcctgagaa gtacaaggaa attttcttcg atcagtctaa aaatggctac
     3601 gccggataca ttgacggcgg agcaagccag gaggaatttt acaaatttat taagcccatc
     3661 ttggaaaaaa tggacggcac cgaggagctg ctggtaaagc ttaacagaga agatctgttg
     3721 cgcaaacagc gcactttcga caatggaagc atcccccacc agattcacct gggcgaactg
     3781 cacgctatcc tcaggcggca agaggatttc tacccctttt tgaaagataa cagggaaaag
     3841 attgagaaaa tcctcacatt tcggataccc tactatgtag gccccctcgc ccggggaaat
     3901 tccagattcg cgtggatgac tcgcaaatca gaagagacca tcactccctg gaacttcgag
     3961 gaagtcgtgg ataagggggc ctctgcccag tccttcatcg aaaggatgac taactttgat
     4021 aaaaatctgc ctaacgaaaa ggtgcttcct aaacactctc tgctgtacga gtacttcaca
     4081 gtttataacg agctcaccaa ggtcaaatac gtcacagaag ggatgagaaa gccagcattc
     4141 ctgtctggag agcagaagaa agctatcgtg gacctcctct tcaagacgaa ccggaaagtt
     4201 accgtgaaac agctcaaaga agactatttc aaaaagattg aatgtttcga ctctgttgaa
     4261 atcagcggag tggaggatcg cttcaacgca tccctgggaa cgtatcacga tctcctgaaa
     4321 atcattaaag acaaggactt cctggacaat gaggagaacg aggacattct tgaggacatt
     4381 gtcctcaccc ttacgttgtt tgaagatagg gagatgattg aagaacgctt gaaaacttac
     4441 gctcatctct tcgacgacaa agtcatgaaa cagctcaaga ggcgccgata tacaggatgg
     4501 gggcggctgt caagaaaact gatcaatggg atccgagaca agcagagtgg aaagacaatc
     4561 ctggattttc ttaagtccga tggatttgcc aaccggaact tcatgcagtt gatccatgat
     4621 gactctctca cctttaagga ggacatccag aaagcacaag tttctggcca gggggacagt
     4681 cttcacgagc acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg aatactgcag
     4741 accgttaagg tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc cgagaatatc
     4801 gttatcgaga tggcccgaga gaaccaaact acccagaagg gacagaagaa cagtagggaa
     4861 aggatgaaga ggattgaaga gggtataaaa gaactggggt cccaaatcct taaggaacac
     4921 ccagttgaaa acacccagct tcagaatgag aagctctacc tgtactacct gcagaacggc
     4981 agggacatgt acgtggatca ggaactggac atcaatcggc tctccgacta cgacgtggat
     5041 catatcgtgc cccagtcttt tctcaaagat gattctattg ataataaagt gttgacaaga
     5101 tccgataaaa atagagggaa gagtgataac gtcccctcag aagaagttgt caagaaaatg
     5161 aaaaattatt ggcggcagct gctgaacgcc aaactgatca cacaacggaa gttcgataat
     5221 ctgactaagg ctgaacgagg tggcctgtct gagttggata aagccggctt catcaaaagg
     5281 cagcttgttg agacacgcca gatcaccaag cacgtggccc aaattctcga ttcacgcatg
     5341 aacaccaagt acgatgaaaa tgacaaactg attcgagagg tgaaagttat tactctgaag
     5401 tctaagctgg tctcagattt cagaaaggac tttcagtttt ataaggtgag agagatcaac
     5461 aattaccacc atgcgcatga tgcctacctg aatgcagtgg taggcactgc acttatcaaa
     5521 aaatatccca agcttgaatc tgaatttgtt tacggagact ataaagtgta cgatgttagg
     5581 aaaatgatcg caaagtctga gcaggaaata ggcaaggcca ccgctaagta cttcttttac
     5641 agcaatatta tgaatttttt caagaccgag attacactgg ccaatggaga gattcggaag
     5701 cgaccactta tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa gggtagggat
     5761 ttcgcgacag tccggaaggt cctgtccatg ccgcaggtga acatcgttaa aaagaccgaa
     5821 gtacagaccg gaggcttctc caaggaaagt atcctcccga aaaggaacag cgacaagctg
     5881 atcgcacgca aaaaagattg ggaccccaag aaatacggcg gattcgattc tcctacagtc
     5941 gcttacagtg tactggttgt ggccaaagtg gagaaaggga agtctaaaaa actcaaaagc
     6001 gtcaaggaac tgctgggcat cacaatcatg gagcgatcaa gcttcgaaaa aaaccccatc
     6061 gactttctcg aggcgaaagg atataaagag gtcaaaaaag acctcatcat taagcttccc
     6121 aagtactctc tctttgagct tgaaaacggc cggaaacgaa tgctcgctag tgcgggcgag
     6181 ctgcagaaag gtaacgagct ggcactgccc tctaaatacg ttaatttctt gtatctggcc
     6241 agccactatg aaaagctcaa agggtctccc gaagataatg agcagaagca gctgttcgtg
     6301 gaacaacaca aacactacct tgatgagatc atcgagcaaa taagcgaatt ctccaaaaga
     6361 gtgatcctcg ccgacgctaa cctcgataag gtgctttctg cttacaataa gcacagggat
     6421 aagcccatca gggagcaggc agaaaacatt atccacttgt ttactctgac caacttgggc
     6481 gcgcctgcag ccttcaagta cttcgacacc accatagaca gaaagcggta cacctctaca
     6541 aaggaggtcc tggacgccac actgattcat cagtcaatta cggggctcta tgaaacaaga
     6601 atcgacctct ctcagctcgg tggagacagc agggctgacc ccaagaagaa gaggaaggtg
     6661 tgaaagggtt cgatccctac cggttagtaa tgagtttaaa cgggggaggc taactgaaac
     6721 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     6781 aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     6841 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     6901 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     6961 gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta
     7021 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     7081 gcgccctagc gcccggggtt ataattacct caggtcgacg tcccatgtgc aggtgctgaa
     7141 ttcgaaggca ttccacgacg tggtcaccag ccatggccat gtccaactcc atcaaatcag
     7201 cttgagtagc catgcccatg gctccagcct gtcctcgtac cagcttttgt tccctttagt
     7261 gagggttaat ttcgagcttg gcgtaatcat ggtcatagct gtttccaatt cttgaagacg
     7321 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     7381 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     7441 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     7501 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     7561 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     7621 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     7681 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     7741 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     7801 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     7861 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     7921 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     7981 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     8041 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     8101 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     8161 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     8221 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     8281 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     8341 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     8401 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     8461 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     8521 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     8581 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     8641 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     8701 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     8761 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     8821 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     8881 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     8941 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     9001 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     9061 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     9121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     9181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     9241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     9301 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     9361 ttcacaccgc atagtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa
     9421 ttggagcttg gcgtaatcat ggtcat
//
