LOCUS       sequence-181553-        9251 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses 3xFLAG-Cas9 and a gRNA targeting mouse Suz12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9251)
  AUTHORS   Vo BT, Li C, Morgan MA, Theurillat I, Finkelstein D, Wright S, Hyle 
            J, Smith SM, Fan Y, Wang YD, Wu G, Orr BA, Northcott PA, Shilatifard
            A, Sherr CJ, Roussel MF
  TITLE     Inactivation of Ezh2 Upregulates Gfi1 and Drives Aggressive 
            Myc-Driven Group 3 Medulloblastoma.
  JOURNAL   Cell Rep. 2017 Mar 21;18(12):2907-2917. doi: 
            10.1016/j.celrep.2017.02.073.
  PUBMED    28329683
REFERENCE   2  (bases 1 to 9251)
  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.celrep.2017.02"; journalName: "Cell Rep"; date: 
            "2017-03-21- 21"; volume: "18"; issue: "12"; pages: "2907-2917"
COMMENT     Sequence Label: AOI-WT-Cas9-sg-mouse Suz12-F1-GFP
FEATURES             Location/Qualifiers
     source          1..9251
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          122..349
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      361..370
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             370..435
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             442..462
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             487..4587
                     /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             487..4587
                     /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             4588..4635
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4651..4704
                     /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(4705..4707,4708..5418)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4705..4707,4708..5418)
                     /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(5008..5027)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5355..5376
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5446..5463)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5452..5659
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5668..5808
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5668..5797
                     /label=AAV2 ITR
     rep_origin      5883..6338
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5970..5989)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6180..6201
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6355..6374)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6474..6496
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6534..6552)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6620..6724
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6725..6793,6794..7585)
                     /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(6943..6962)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7756..8344
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8245..8264
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8406..8654
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8406..8426
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8577..8596
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8676..8751
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8848..9133
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
       61 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      121 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      181 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      241 tccgggctgt aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt
      301 attaatgttt aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt
      361 gccaccatgg actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa
      421 gacgatgacg ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca
      481 gcagccgaca agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc
      541 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac
      601 cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc
      661 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc
      721 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
      781 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc
      841 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga
      901 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc
      961 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
     1021 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac
     1081 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc
     1141 agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga
     1201 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
     1261 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
     1321 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc
     1381 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc
     1441 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg
     1501 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac
     1561 gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc
     1621 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg
     1681 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg
     1741 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag
     1801 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac
     1861 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag
     1921 gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat
     1981 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
     2041 gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc
     2101 ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg
     2161 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa
     2221 atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa
     2281 attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc
     2341 gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat
     2401 gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg
     2461 ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc
     2521 ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac
     2581 gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc
     2641 ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag
     2701 acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc
     2761 gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag
     2821 agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac
     2881 cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg
     2941 cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac
     3001 catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga
     3061 agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg
     3121 aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat
     3181 ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga
     3241 cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg
     3301 aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag
     3361 tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac
     3421 aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa
     3481 aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg
     3541 aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac
     3601 agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag
     3661 cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat
     3721 tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag
     3781 gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg
     3841 atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg
     3901 gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt
     3961 gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc
     4021 gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct
     4081 aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa
     4141 ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc
     4201 agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg
     4261 gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga
     4321 gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat
     4381 aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga
     4441 gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc
     4501 aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg
     4561 atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag
     4621 gcaaaaaaga aaaaggaatt cggcagtgga gagggcagag gaagtctgct aacatgcggt
     4681 gacgtcgagg agaatcctgg cccagtgagc aagggcgagg agctgttcac cggggtggtg
     4741 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     4801 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     4861 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     4921 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     4981 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     5041 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     5101 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     5161 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     5221 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     5281 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     5341 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     5401 atggacgagc tgtacaagga attctaacta gagctcgctg atcagcctcg actgtgcctt
     5461 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     5521 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     5581 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga
     5641 atagcaggca tgctggggag cggccgcagg aacccctagt gatggagttg gccactccct
     5701 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     5761 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     5821 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     5881 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5941 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     6001 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     6061 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     6121 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     6181 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     6241 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     6301 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     6361 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     6421 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     6481 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     6541 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     6601 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6661 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6721 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6781 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6841 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6901 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6961 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     7021 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     7081 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     7141 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7201 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7261 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7321 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7381 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7441 aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7501 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7561 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7621 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7681 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7741 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7801 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7861 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     7921 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7981 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     8041 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     8101 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     8161 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     8221 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     8281 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     8341 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     8401 catgtgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
     8461 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
     8521 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
     8581 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
     8641 aaggacgaaa caccgcggct tcgggggttc ggcgggtttt agagctagaa atagcaagtt
     8701 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt
     8761 ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt
     8821 ctgcagacaa atggctctag aggtacccgt tacataactt acggtaaatg gcccgcctgg
     8881 ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca
     8941 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     9001 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg
     9061 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     9121 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
     9181 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
     9241 gggggggggg g
//
