LOCUS       sequence-492882-        8505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpCas9 and a sgRNA targeting the human AAVS1 loci for 
            knock-in..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8505)
  AUTHORS   Ide S, Imai R, Ochi H, Maeshima K
  TITLE     Transcriptional suppression of ribosomal DNA with phase separation.
  JOURNAL   Sci Adv. 2020 Oct 14;6(42). pii: 6/42/eabb5953. doi: 
            10.1126/sciadv.abb5953. Print 2020 Oct.
  PUBMED    33055158
REFERENCE   2  (bases 1 to 8505)
  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"; journalName: "Sci Adv."; 
            date: "2020-10-14"; volume: "6(42). pii: 6/42/eabb5953. doi"; pages:
            " 10.1126/sciadv.abb5953. Print 2020 Oct"
COMMENT     Sequence Label: pX330-AAVS1
FEATURES             Location/Qualifiers
     source          1..8505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     79..98
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   260..400
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   271..400
                     /label=AAV2 ITR
     polyA_signal    409..616
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     605..622
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(650..697)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(698..4798)
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(698..4798)
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(4823..4843)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4850..4915)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      4915..4924
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          4936..5163
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     promoter        complement(5164..5439)
                     /label=chicken beta-actin promoter
     enhancer        5441..5726
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     misc_RNA        complement(5823..5898)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(5919..6167)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(5977..5996)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(6147..6167)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(6229..6817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6309..6328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6988..7779,7780..7848))
                     /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     7611..7630
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7849..7953)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8021..8039
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8077..8099)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8199..8218
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(8235..8505,1..185))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8372..8393)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
       61 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag
      121 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
      181 cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga
      241 aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
      301 cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
      361 cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgctc cccagcatgc
      421 ctgctattct cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata
      481 gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg
      541 ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg
      601 caactagaag gcacagtcga ggctgatcag cgagctctag gaattcttac tttttctttt
      661 ttgcctggcc ggcctttttc gtggccgccg gccttttgtc gcctcccagc tgagacaggt
      721 cgatccgtgt ctcgtacagg ccggtgatgc tctggtggat cagggtggcg tccagcacct
      781 ctttggtgct ggtgtacctc ttccggtcga tggtggtgtc aaagtacttg aaggcggcag
      841 gggctcccag attggtcagg gtaaacaggt ggatgatatt ctcggcctgc tctctgatgg
      901 gcttatcccg gtgcttgttg taggcggaca gcactttgtc cagattagcg tcggccagga
      961 tcactctctt ggagaactcg ctgatctgct cgatgatctc gtccaggtag tgcttgtgct
     1021 gttccacaaa cagctgtttc tgctcattat cctcggggga gcccttcagc ttctcatagt
     1081 ggctggccag gtacaggaag ttcacatatt tggagggcag ggccagttcg tttcccttct
     1141 gcagttcgcc ggcagaggcc agcattctct tccggccgtt ttccagctcg aacagggagt
     1201 acttaggcag cttgatgatc aggtcctttt tcacttcttt gtagcccttg gcttccagaa
     1261 agtcgatggg attcttctcg aagctgcttc tttccatgat ggtgatcccc agcagctctt
     1321 tcacactctt cagtttcttg gacttgccct tttccacttt ggccaccacc agcacagaat
     1381 aggccacggt ggggctgtcg aagccgccgt acttcttagg gtcccagtcc ttctttctgg
     1441 cgatcagctt atcgctgttc ctcttgggca ggatagactc tttgctgaag ccgcctgtct
     1501 gcacctcggt ctttttcacg atattcactt ggggcatgct cagcactttc cgcacggtgg
     1561 caaaatcccg gcccttatcc cacacgatct ccccggtttc gccgtttgtc tcgatcagag
     1621 gccgcttccg gatctcgccg ttggccaggg taatctcggt cttgaaaaag ttcatgatgt
     1681 tgctgtagaa gaagtacttg gcggtagcct tgccgatttc ctgctcgctc ttggcgatca
     1741 tcttccgcac gtcgtacacc ttgtagtcgc cgtacacgaa ctcgctttcc agcttagggt
     1801 actttttgat cagggcggtt cccacgacgg cgttcaggta ggcgtcgtgg gcgtggtggt
     1861 agttgttgat ctcgcgcact ttgtaaaact ggaaatcctt ccggaaatcg gacaccagct
     1921 tggacttcag ggtgatcact ttcacttccc ggatcagctt gtcattctcg tcgtacttag
     1981 tgttcatccg ggagtccagg atctgtgcca cgtgctttgt gatctgccgg gtttccacca
     2041 gctgtctctt gatgaagccg gccttatcca gttcgctcag gccgcctctc tcggccttgg
     2101 tcagattgtc gaactttctc tgggtaatca gcttggcgtt cagcagctgc cgccagtagt
     2161 tcttcatctt cttcacgacc tcttcggagg gcacgttgtc gctcttgccc cggttcttgt
     2221 cgcttctggt cagcaccttg ttgtcgatgg agtcgtcctt cagaaagctc tgaggcacga
     2281 tatggtccac atcgtagtcg gacagccggt tgatgtccag ttcctggtcc acgtacatat
     2341 cccgcccatt ctgcaggtag tacaggtaca gcttctcgtt ctgcagctgg gtgttttcca
     2401 cggggtgttc tttcaggatc tggctgccca gctctttgat gccctcttcg atccgcttca
     2461 ttctctcgcg gctgttcttc tgtcccttct gggtggtctg gttctctctg gccatttcga
     2521 tcacgatgtt ctcgggcttg tgccggccca tcactttcac gagctcgtcc accaccttca
     2581 ctgtctgcag gatgcccttc ttaatggcgg ggctgccggc cagattggca atgtgctcgt
     2641 gcaggctatc gccctggccg gacacctggg ctttctggat gtcctcttta aaggtcaggc
     2701 tgtcgtcgtg gatcagctgc atgaagtttc tgttggcgaa gccgtcggac ttcaggaaat
     2761 ccaggattgt cttgccggac tgcttgtccc ggatgccgtt gatcagcttc cggctcagcc
     2821 tgccccagcc ggtgtatctc cgccgcttca gctgcttcat cactttgtcg tcgaacaggt
     2881 gggcataggt tttcagccgt tcctcgatca tctctctgtc ctcaaacagt gtcagggtca
     2941 gcacgatatc ttccagaatg tcctcgtttt cctcattgtc caggaagtcc ttgtccttga
     3001 taattttcag cagatcgtgg tatgtgccca gggaggcgtt gaaccgatct tccacgccgg
     3061 agatttccac ggagtcgaag cactcgattt tcttgaagta gtcctctttc agctgcttca
     3121 cggtcacttt ccggttggtc ttgaacagca ggtccacgat ggcctttttc tgctcgccgc
     3181 tcaggaaggc gggctttctc attccctcgg tcacgtattt cactttggtc agctcgttat
     3241 acacggtgaa gtactcgtac agcaggctgt gcttgggcag caccttctcg ttgggcaggt
     3301 tcttatcgaa gttggtcatc cgctcgatga agctctgggc ggaagcgccc ttgtccacca
     3361 cttcctcgaa gttccagggg gtgatggttt cctcgctctt tctggtcatc caggcgaatc
     3421 tgctgtttcc cctggccaga gggcccacgt agtaggggat gcggaaggtc aggatcttct
     3481 cgatcttttc ccggttgtcc ttcaggaatg ggtaaaaatc ttcctgccgc cgcagaatgg
     3541 cgtgcagctc tcccaggtgg atctggtggg ggatgctgcc gttgtcgaag gtccgctgct
     3601 tccgcagcag gtcctctctg ttcagcttca cgagcagttc ctcggtgccg tccatctttt
     3661 ccaggatggg cttgatgaac ttgtagaact cttcctggct ggctccgccg tcaatgtagc
     3721 cggcgtagcc gttcttgctc tggtcgaaga aaatctcttt gtacttctca ggcagctgct
     3781 gccgcacgag agctttcagc agggtcaggt cctggtggtg ctcgtcgtat ctcttgatca
     3841 tagaggcgct caggggggcc ttggtgatct cggtgttcac tctcaggatg tcgctcagca
     3901 ggatggcgtc ggacaggttc ttggcggcca gaaacaggtc ggcgtactgg tcgccgatct
     3961 gggccagcag gttgtccagg tcgtcgtcgt aggtgtcctt gctcagctgc agtttggcat
     4021 cctcggccag gtcgaagttg ctcttgaagt tgggggtcag gcccaggctc agggcaatca
     4081 ggtttccgaa caggccattc ttcttctcgc cgggcagctg ggcgatcaga ttttccagcc
     4141 gtctgctctt gctcagtctg gcagacagga tggccttggc gtccacgccg ctggcgttga
     4201 tggggttttc ctcgaacagc tggttgtagg tctgcaccag ctggatgaac agcttgtcca
     4261 cgtcgctgtt gtcggggttc aggtcgccct cgatcaggaa gtggccccgg aacttgatca
     4321 tgtgggccag ggccagatag atcagccgca ggtcggcctt gtcggtgctg tccaccagtt
     4381 tctttctcag gtggtagatg gtggggtact tctcgtggta ggccacctcg tccacgatgt
     4441 tgccgaagat ggggtgccgc tcgtgcttct tatcctcttc caccaggaag gactcttcca
     4501 gtctgtggaa gaagctgtcg tccaccttgg ccatctcgtt gctgaagatc tcttgcagat
     4561 agcagatccg gttcttccgt ctggtgtatc ttcttctggc ggttctcttc agccgggtgg
     4621 cctcggctgt ttcgccgctg tcgaacagca gggctccgat caggttcttc ttgatgctgt
     4681 gccggtcggt gttgcccagc accttgaatt tcttgctggg caccttgtac tcgtcggtga
     4741 tcacggccca gcccacagag ttggtgccga tgtccaggcc gatgctgtac ttcttgtcgg
     4801 ctgctgggac tccgtggata ccgaccttcc gcttcttctt tggggccatc ttatcgtcat
     4861 cgtctttgta atcaatatca tgatccttgt agtctccgtc gtggtcctta tagtccatgg
     4921 tggcaccggt ccaacctgaa aaaaagtgat ttcaggcagg tgctccaggt aattaaacat
     4981 taatacccca ccaaccaacc atcccttaaa cccttacctc ttgctcagct aattacagcc
     5041 cggaggagaa gggccgtccc gcccgctcac ctgtgggagt aacgcggtca gtcagagccg
     5101 gggcgggcgg cgcgaggcgg cggcggagcg gggcacgggg cgaaggcagc gcgcagcgac
     5161 tcccgcccgc cgcgcgcttc gctttttata gggccgccgc cgccgccgcc tcgccataaa
     5221 aggaaacttt cggagcgcgc cgctctgatt ggctgccgcc gcacctctcc gcctcgcccc
     5281 gccccgcccc tcgccccgcc ccgccccgcc tggcgcgccc cccccccccc cccgccccca
     5341 tcgctgcaca aaataattaa aaaataaata aatacaaaat tgggggtggg gagggggggg
     5401 agatggggag agtgaagcag aacgtggggc tcacctcgac catggtaata gcgatgacta
     5461 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcacaatgcc
     5521 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
     5581 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
     5641 tccctattgg cgttactatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     5701 ggccatttac cgtaagttat gtaacgggta cctctagagc catttgtctg cagaattggc
     5761 gcacgcgcta aaaacggact agccttattt taacttgcta tttctagctc taaaacaaaa
     5821 aagcaccgac tcggtgccac tttttcaagt tgataacgga ctagccttat tttaacttgc
     5881 tatttctagc tctaaaacat cctgtcccta gtggccccgg tgtttcgtcc tttccacaag
     5941 atatataaag ccaagaaatc gaaatacttt caagttacgg taagcatatg atagtccatt
     6001 ttaaaacata attttaaaac tgcaaactac ccaagaaatt attactttct acgtcacgta
     6061 ttttgtacta atatctttgt gtttacagtc aaattaattc caattatctc tctaacagcc
     6121 ttgtatcgta tatgcaaata tgaaggaatc atgggaaata ggccctcaca tgtgagcaaa
     6181 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6241 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6301 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6361 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6421 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6481 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6541 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6601 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6661 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6721 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6781 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6841 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6901 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6961 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7021 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7081 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggc ttccacgctc
     7141 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7201 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7261 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7321 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7381 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7441 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7501 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7561 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7621 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7681 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7741 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7801 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7861 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7921 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7981 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     8041 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     8101 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     8161 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     8221 gagagtgcac cataaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
     8281 ttaaatcagc tcatttttta accaatagac cgaaatcggc aaaatccctt ataaatcaaa
     8341 agaatagccc gagatagagt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     8401 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     8461 tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc cgtaa
//
