LOCUS       sequence-183214-        9129 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses 3xFLAG-Cas9 and a gRNA targeting mouse Gfi1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9129)
  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 9129)
  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 Gfi1-exon5 (F3)-GFP
FEATURES             Location/Qualifiers
     source          1..9129
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          6..233
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      245..254
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             254..319
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             326..346
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             371..4471
                     /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             371..4471
                     /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             4472..4519
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4535..4588
                     /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(4589..4591,4592..5302)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4589..4591,4592..5302)
                     /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(4892..4911)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5239..5260
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5330..5347)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5336..5543
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5552..5692
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5552..5681
                     /label=AAV2 ITR
     rep_origin      5767..6222
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5854..5873)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6064..6085
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6239..6258)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6358..6380
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6418..6436)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6504..6608
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6609..6677,6678..7469)
                     /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(6827..6846)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7640..8228
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8129..8148
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8290..8538
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8290..8310
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8461..8480
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8560..8635
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8732..9017
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
       61 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
      121 ctcctccggg ctgtaattag ctgagcaaga ggtaagggtt taagggatgg ttggttggtg
      181 gggtattaat gtttaattac ctggagcacc tgcctgaaat cacttttttt caggttggac
      241 cggtgccacc atggactata aggaccacga cggagactac aaggatcatg atattgatta
      301 caaagacgat gacgataaga tggccccaaa gaagaagcgg aaggtcggta tccacggagt
      361 cccagcagcc gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg
      421 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
      481 cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac
      541 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
      601 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
      661 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
      721 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
      781 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
      841 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
      901 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
      961 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
     1021 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     1081 cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     1141 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     1201 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     1261 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     1321 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     1381 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     1441 ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     1501 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     1561 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     1621 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     1681 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     1741 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     1801 cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt
     1861 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     1921 caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     1981 cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa
     2041 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     2101 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     2161 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     2221 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     2281 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     2341 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     2401 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     2461 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     2521 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     2581 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     2641 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
     2701 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
     2761 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     2821 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
     2881 ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac
     2941 cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
     3001 gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga
     3061 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
     3121 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
     3181 gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct
     3241 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
     3301 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
     3361 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
     3421 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     3481 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     3541 gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg
     3601 ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac
     3661 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa
     3721 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
     3781 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     3841 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     3901 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     3961 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
     4021 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     4081 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
     4141 tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa
     4201 gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg
     4261 ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
     4321 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
     4381 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
     4441 acggatcgac ctgtctcagc tgggaggcga caaaaggccg gcggccacga aaaaggccgg
     4501 ccaggcaaaa aagaaaaagg aattcggcag tggagagggc agaggaagtc tgctaacatg
     4561 cggtgacgtc gaggagaatc ctggcccagt gagcaagggc gaggagctgt tcaccggggt
     4621 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     4681 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     4741 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     4801 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     4861 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     4921 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     4981 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     5041 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     5101 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     5161 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     5221 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     5281 cggcatggac gagctgtaca aggaattcta actagagctc gctgatcagc ctcgactgtg
     5341 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     5401 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     5461 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     5521 gagaatagca ggcatgctgg ggagcggccg caggaacccc tagtgatgga gttggccact
     5581 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     5641 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     5701 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     5761 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5821 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     5881 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5941 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     6001 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     6061 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     6121 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     6181 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     6241 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     6301 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     6361 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     6421 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     6481 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6541 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6601 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6661 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6721 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6781 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6841 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6901 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6961 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     7021 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     7081 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7141 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     7201 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     7261 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7321 gtggaagccg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7381 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7441 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     7501 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     7561 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7621 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     7681 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     7741 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     7801 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     7861 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     7921 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     7981 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     8041 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     8101 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     8161 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     8221 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     8281 ctcacatgtg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
     8341 gttagagaga taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg
     8401 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
     8461 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
     8521 tggaaaggac gaaacaccgc ttcgggcacg cggtgagccg ttttagagct agaaatagca
     8581 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     8641 tgttttagag ctagaaatag caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc
     8701 aattctgcag acaaatggct ctagaggtac ccgttacata acttacggta aatggcccgc
     8761 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat agtaacgcca atagggactt
     8821 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     8881 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     8941 attgtgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     9001 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
     9061 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
     9121 gggcggggg
//
