LOCUS       sequence-273921-        9715 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  gRNA to knockout EED. Use with EED-KO-gRNA-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9715)
  AUTHORS   Oksuz O, Narendra V, Lee CH, Descostes N, LeRoy G, Raviram R, 
            Blumenberg L, Karch K, Rocha PP, Garcia BA, Skok JA, Reinberg D
  TITLE     Capturing the Onset of PRC2-Mediated Repressive Domain Formation.
  JOURNAL   Mol Cell. 2018 Jun 21;70(6):1149-1162.e5. doi: 
            10.1016/j.molcel.2018.05.023.
  PUBMED    29932905
REFERENCE   2  (bases 1 to 9715)
  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.2018.05"; journalName: "Mol Cell"; date: 
            "2018-06-21- 21"; volume: "70"; issue: "6"; pages: "1149-1162.e5"
COMMENT     Sequence Label: EED-KO-gRNA-1
FEATURES             Location/Qualifiers
     source          1..9715
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          118..345
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      357..366
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             366..431
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             438..458
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             483..4583
                     /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             483..4583
                     /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             4584..4631
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4647..4700
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(4701..4703,4704..5414)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4701..4703,4704..5414)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5004..5023)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5351..5372
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5442..5459)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5448..5655
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5664..5804
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5664..5793
                     /label=AAV2 ITR
     rep_origin      5879..6334
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5966..5985)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6176..6197
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6351..6370)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6470..6492
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6530..6548)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6616..6720
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6721..6789,6790..7581)
                     /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(6939..6958)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7752..8340
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8241..8260
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8402..8650
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8402..8422
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8573..8592
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8672..8747
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8844..9129
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
       61 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
      121 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
      181 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
      241 ggctgtaatt agctgagcaa gaggtaaggg tttaagggat ggttggttgg tggggtatta
      301 atgtttaatt acctggagca cctgcctgaa atcacttttt ttcaggttgg accggtgcca
      361 ccatggacta taaggaccac gacggagact acaaggatca tgatattgat tacaaagacg
      421 atgacgataa gatggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag
      481 ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga
      541 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
      601 acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg
      661 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
      721 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
      781 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
      841 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
      901 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
      961 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     1021 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     1081 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     1141 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc
     1201 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     1261 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     1321 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     1381 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     1441 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     1501 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     1561 gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     1621 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     1681 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     1741 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     1801 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     1861 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     1921 tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     1981 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     2041 ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     2101 gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     2161 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     2221 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     2281 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     2341 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     2401 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     2461 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     2521 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     2581 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     2641 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     2701 tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga
     2761 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     2821 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     2881 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     2941 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata
     3001 tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg
     3061 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     3121 actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga
     3181 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     3241 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
     3301 ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca
     3361 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
     3421 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
     3481 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
     3541 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
     3601 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
     3661 ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg
     3721 ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc
     3781 agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg
     3841 ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct
     3901 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
     3961 aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
     4021 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
     4081 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc
     4141 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
     4201 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
     4261 agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
     4321 tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc
     4381 ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc
     4441 ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag
     4501 aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg
     4561 acctgtctca gctgggaggc gacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa
     4621 aaaagaaaaa ggaattcggc agtggagagg gcagaggaag tctgctaaca tgcggtgacg
     4681 tcgaggagaa tcctggccca gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     4741 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     4801 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     4861 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     4921 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     4981 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     5041 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     5101 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     5161 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     5221 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     5281 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     5341 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     5401 acgagctgta caaggaattc taactagagc tcgctgatca gcctcgactg tgccttctag
     5461 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     5521 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     5581 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag
     5641 caggcatgct ggggagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     5701 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     5761 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     5821 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     5881 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5941 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     6001 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     6061 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     6121 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     6181 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     6241 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     6301 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     6361 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     6421 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     6481 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     6541 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     6601 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6661 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6721 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6781 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6841 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6901 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6961 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     7021 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     7081 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     7141 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     7201 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     7261 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     7321 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7381 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc
     7441 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     7501 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     7561 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     7621 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7681 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7741 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7801 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7861 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     7921 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7981 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     8041 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     8101 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     8161 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     8221 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     8281 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8341 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8401 tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
     8461 gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
     8521 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
     8581 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
     8641 acgaaacacc gctctggcta ctgtcaacta ggttttagag ctagaaatag caagttaaaa
     8701 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttgttttag
     8761 agctagaaat agcaagttaa aataaggcta gtccgttttt agcgcgtgcg ccaattctgc
     8821 agacaaatgg ctctagaggt acccgttaca taacttacgg taaatggccc gcctggctga
     8881 ccgcccaacg acccccgccc attgacgtca atagtaacgc caatagggac tttccattga
     8941 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     9001 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattgtgcc
     9061 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     9121 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
     9181 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
     9241 gggggggggg ggsgsgccch sgcggggggg gggggggggg gggggggggg gggggggggg
     9301 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
     9361 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
     9421 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
     9481 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
     9541 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
     9601 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
     9661 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg ggggg
//
