LOCUS       sequence-338134-        8475 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses Cas9 and an sgRNA targeting SUZ12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8475)
  AUTHORS   Youmans DT, Schmidt JC, Cech TR
  TITLE     Live-cell imaging reveals the dynamics of PRC2 and recruitment to 
            chromatin by SUZ12-associated subunits.
  JOURNAL   Genes Dev. 2018 Jun 1;32(11-12):794-805. doi: 
            10.1101/gad.311936.118. Epub 2018 Jun 11.
  PUBMED    29891558
REFERENCE   2  (bases 1 to 8475)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genes 
            Dev."; date: "2018-06-1"; volume: "32(11-12):794-805. doi"; pages: "
            10.1101/gad.311936.118. Epub 2018 Jun 11"
COMMENT     Sequence Label: px330 SUZ12 - pDY083
FEATURES             Location/Qualifiers
     source          1..8475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          130..357
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      369..378
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             378..443
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             450..470
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             495..4595
                     /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             495..4595
                     /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             4596..4643
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4671..4688)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4677..4884
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4893..5033
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4893..5022
                     /label=AAV2 ITR
     rep_origin      5108..5563
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5195..5214)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5405..5426
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5580..5599)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5699..5721
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5759..5777)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5845..5949
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5950..6018,6019..6810)
                     /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(6168..6187)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6981..7569
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7470..7489
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7631..7879
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7631..7651
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7802..7821
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7900..7975
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8072..8357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc
       61 cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg
      121 cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg
      181 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc
      241 ccttctcctc cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg atggttggtt
      301 ggtggggtat taatgtttaa ttacctggag cacctgcctg aaatcacttt ttttcaggtt
      361 ggaccggtgc caccatggac tataaggacc acgacggaga ctacaaggat catgatattg
      421 attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc ggtatccacg
      481 gagtcccagc agccgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg
      541 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
      601 acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg
      661 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
      721 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
      781 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
      841 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
      901 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
      961 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
     1021 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
     1081 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
     1141 gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
     1201 tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
     1261 acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
     1321 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
     1381 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
     1441 tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag
     1501 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
     1561 acggctacgc cggctacatt gacggcggag ccagccagga agagttctac aagttcatca
     1621 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg
     1681 acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg
     1741 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
     1801 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
     1861 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
     1921 acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca
     1981 acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
     2041 acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
     2101 ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc
     2161 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
     2221 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
     2281 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
     2341 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
     2401 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
     2461 ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
     2521 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga
     2581 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
     2641 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
     2701 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
     2761 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
     2821 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
     2881 aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
     2941 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
     3001 atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc
     3061 tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
     3121 agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt
     3181 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
     3241 tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact
     3301 cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca
     3361 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
     3421 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc
     3481 tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg
     3541 acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact
     3601 tcttctacag caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga
     3661 tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg
     3721 gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa
     3781 agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg
     3841 ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc
     3901 ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac
     3961 tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga
     4021 atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca
     4081 agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg
     4141 ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt
     4201 acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc
     4261 tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct
     4321 ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc
     4381 accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca
     4441 atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca
     4501 ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg
     4561 agacacggat cgacctgtct cagctgggag gcgacaaaag gccggcggcc acgaaaaagg
     4621 ccggccaggc aaaaaagaaa aagtaagaat tcctagagct cgctgatcag cctcgactgt
     4681 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     4741 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     4801 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4861 agagaatagc aggcatgctg gggagcggcc gcaggaaccc ctagtgatgg agttggccac
     4921 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     4981 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     5041 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     5101 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5161 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     5221 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5281 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     5341 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5401 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     5461 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     5521 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     5581 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     5641 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     5701 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     5761 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5821 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5881 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5941 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6001 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6061 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6121 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6181 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6241 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6301 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6361 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6421 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6481 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6541 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6601 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6661 cgtggaagcc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6721 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6781 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6841 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6901 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6961 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7021 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7081 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7141 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7201 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7261 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7321 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7381 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7441 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7501 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7561 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7621 gctcacatgt gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
     7681 tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
     7741 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
     7801 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
     7861 gtggaaagga cgaaacaccg cctcccccgg accccgcgcg ttttagagct agaaatagca
     7921 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     7981 tgttttagag ctagaaatag caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc
     8041 aattctgcag acaaatggct ctagaggtac ccgttacata acttacggta aatggcccgc
     8101 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat agtaacgcca atagggactt
     8161 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     8221 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     8281 attgtgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     8341 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
     8401 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
     8461 gggcgggggg ggggg
//
