LOCUS       sequence-338121-        8487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid encodes Cas9 and a sgRNA that targets the SUZ12 locus; 
            to be used with pDY153.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8487)
  AUTHORS   Youmans DT, Gooding AR, Dowell RD, Cech TR
  TITLE     Competition between PRC2.1 and 2.2 subcomplexes regulates PRC2 
            chromatin occupancy in human stem cells.
  JOURNAL   Mol Cell. 2021 Feb 4;81(3):488-501.e9. doi: 
            10.1016/j.molcel.2020.11.044. Epub 2020 Dec 17.
  PUBMED    33338397
REFERENCE   2  (bases 1 to 8487)
  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.molcel.2020.11.044"; journalName: "Mol Cell"; date: 
            "2021-02-4- 4"; volume: "81"; issue: "3"; pages: "488-501.e9"
COMMENT     Sequence Label: pDY156: SUZ12 px330 sgRNA Cas9 plasmid
FEATURES             Location/Qualifiers
     source          1..8487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          140..367
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      379..388
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             388..453
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             460..480
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             505..4605
                     /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             505..4605
                     /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             4606..4653
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4681..4698)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4687..4894
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4903..5043
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4903..5032
                     /label=AAV2 ITR
     rep_origin      5118..5573
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5205..5224)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5415..5436
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5590..5609)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5709..5731
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5769..5787)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5855..5959
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5960..6028,6029..6820)
                     /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(6178..6197)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6991..7579
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7480..7499
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7641..7889
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7641..7661
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7812..7831
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7911..7986
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8083..8368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(8370..8487,1..139)
                     /label=chicken beta-actin promoter
                     /note="contains a 21-bp deletion relative to the standard 
                     chicken beta-actin promoter"
ORIGIN
        1 ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag
       61 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
      121 gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc
      181 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
      241 gcgggacggc ccttctcctc cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg
      301 atggttggtt ggtggggtat taatgtttaa ttacctggag cacctgcctg aaatcacttt
      361 ttttcaggtt ggaccggtgc caccatggac tataaggacc acgacggaga ctacaaggat
      421 catgatattg attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc
      481 ggtatccacg gagtcccagc agccgacaag aagtacagca tcggcctgga catcggcacc
      541 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
      601 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
      661 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
      721 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
      781 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
      841 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
      901 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
      961 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     1021 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     1081 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
     1141 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     1201 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     1261 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     1321 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     1381 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     1441 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     1501 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     1561 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
     1621 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     1681 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
     1741 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     1801 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     1861 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     1921 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
     1981 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     2041 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
     2101 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
     2161 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     2221 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     2281 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     2341 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     2401 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     2461 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     2521 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     2581 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     2641 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     2701 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     2761 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     2821 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     2881 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     2941 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     3001 tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac
     3061 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     3121 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
     3181 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     3241 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     3301 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
     3361 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     3421 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     3481 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     3541 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     3601 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     3661 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
     3721 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
     3781 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
     3841 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
     3901 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
     3961 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
     4021 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
     4081 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
     4141 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
     4201 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
     4261 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
     4321 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
     4381 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
     4441 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
     4501 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
     4561 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaaaag gccggcggcc
     4621 acgaaaaagg ccggccaggc aaaaaagaaa aagtaagaat tcctagagct cgctgatcag
     4681 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     4741 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     4801 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     4861 aggattggga agagaatagc aggcatgctg gggagcggcc gcaggaaccc ctagtgatgg
     4921 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4981 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     5041 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     5101 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5161 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     5221 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5281 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     5341 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5401 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     5461 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     5521 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     5581 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     5641 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5701 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5761 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5821 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5881 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5941 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     6001 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     6061 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     6121 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     6181 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6241 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6301 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6361 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6421 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6481 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6541 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6601 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6661 agccggtgag cgtggaagcc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6721 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6781 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6841 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6901 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6961 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     7021 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     7081 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     7141 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     7201 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     7261 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7321 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7381 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7441 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7501 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7561 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7621 ctggcctttt gctcacatgt gagggcctat ttcccatgat tccttcatat ttgcatatac
     7681 gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag
     7741 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
     7801 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
     7861 tatatatctt gtggaaagga cgaaacaccg tctatagatt tcttcgaact gttttagagc
     7921 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
     7981 cggtgctttt ttgttttaga gctagaaata gcaagttaaa ataaggctag tccgttttta
     8041 gcgcgtgcgc caattctgca gacaaatggc tctagaggta cccgttacat aacttacggt
     8101 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc
     8161 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     8221 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     8281 gcccgcctgg cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     8341 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
     8401 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
     8461 tgcagcgatg ggggcggggg ggggggg
//
