LOCUS       sequence-466603-       11687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable knockout of DNMT3B function in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11687)
  AUTHORS   Choudhury SR, Kaushal A, Biswas P, Padilla C, Sarthy JF, Chavan A, 
            Gonzalez GA, Meshinchi S, Farrar JE
  TITLE     Transcriptional rewiring by enhancer methylation in 
            CBFA2T3-GLIS2-driven pediatric acute megakaryoblastic leukemia.
  JOURNAL   Genes Dis. 2025 Sep 6;13(1):101843. doi: 
            10.1016/j.gendis.2025.101843. eCollection 2026 Jan.
  PUBMED    41141393
REFERENCE   2  (bases 1 to 11687)
  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"; journalName: "Genes 
            Dis."; date: "2025-09-6"; volume: "13(1):101843. doi"; pages: " 
            10.1016/j.gendis.2025.101843. eCollection 2026 Jan"
COMMENT     Sequence Label: hDNMT3B_KO_puro
FEATURES             Location/Qualifiers
     source          1..11687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    135..260
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    753..986
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1171..1215
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1481..1598
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1605..1853
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1605..1625
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1776..1795
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1875..1950
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    1976..1996
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        2016..2227
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     protein_bind    2228..2252
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2253..2262
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2262..2327
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2334..2354
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2379..6479
                     /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             2379..6479
                     /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             6480..6527
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6537..6590
                     /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             6591..7190
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6591..6610)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7087..7107
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(7191..7215)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    7226..7814
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7279..7299)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7697..7708)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7886..8119
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8191..8312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8228..8247)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8282..8301
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8352..8487
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8507..8526)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8508..8526)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8536..8553)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8536..8552)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8545..8567)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8694..9149
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8781..8800)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8991..9012
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9175..9279
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9280..9348,9349..10140)
                     /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(9498..9517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10311..10899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10800..10819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11053..11070
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11187..11208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11223..11228,11229..11246,11247..11253)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11261..11277
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     11266..11288
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11285..11301
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11285..11301
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11320..11340
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11322..11340
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        11368..11594
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             join(11595..11687,1..88)
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
       61 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
      121 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
      181 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
      241 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
      301 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
      361 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
      421 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
      481 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
      541 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
      601 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
      661 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
      721 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
      781 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
      841 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
      901 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
      961 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     1021 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     1081 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     1141 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     1201 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     1261 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     1321 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     1381 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
     1441 tccattcgat tagtgaacgg atctcgacgg tatcgctagc ttttaaaaga aaagggggga
     1501 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     1561 aagaattaca aaaacaaatt acaaaaattc aaaattttac tagtgagggc ctatttccca
     1621 tgattccttc atatttgcat atacgataca aggctgttag agagataatt ggaattaatt
     1681 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
     1741 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     1801 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accgtgctgt
     1861 tgcccgccgt ctcagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
     1921 caacttgaaa aagtggcacc gagtcggtgc ttttttgtct agaggtaccg aattccaact
     1981 ttgtatagaa aagttgggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     2041 ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg
     2101 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     2161 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     2221 aacacaggca agtttgtaca aaaaagcagg ctgccaccat ggactataag gaccacgacg
     2281 gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg gccccaaaga
     2341 agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac agcatcggcc
     2401 tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     2461 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
     2521 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
     2581 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
     2641 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
     2701 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
     2761 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
     2821 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
     2881 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     2941 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     3001 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
     3061 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
     3121 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
     3181 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     3241 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     3301 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
     3361 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     3421 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
     3481 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
     3541 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
     3601 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
     3661 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
     3721 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
     3781 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
     3841 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
     3901 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
     3961 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
     4021 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
     4081 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
     4141 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
     4201 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
     4261 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
     4321 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
     4381 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
     4441 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
     4501 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
     4561 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
     4621 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
     4681 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
     4741 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
     4801 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
     4861 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg
     4921 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
     4981 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
     5041 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
     5101 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
     5161 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
     5221 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
     5281 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
     5341 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
     5401 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
     5461 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
     5521 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
     5581 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
     5641 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
     5701 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
     5761 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
     5821 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
     5881 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
     5941 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
     6001 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
     6061 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
     6121 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
     6181 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
     6241 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
     6301 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
     6361 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
     6421 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca
     6481 aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagggc tccggagagg
     6541 gcaggggaag tcttctaaca tgcggggacg tggaggaaaa tcccggcccc atgaccgagt
     6601 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     6661 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     6721 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     6781 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     6841 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     6901 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     6961 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     7021 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     7081 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     7141 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga acccagcttt
     7201 cttgtacaaa gtggtggtac ccgataatca acctctggat tacaaaattt gtgaaagatt
     7261 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     7321 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     7381 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     7441 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     7501 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     7561 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     7621 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     7681 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     7741 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     7801 ggccgcctcc ccgcatcggc tttaagacca atgacttaca aggcagctgt agatcttagc
     7861 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     7921 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     7981 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     8041 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     8101 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     8161 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     8221 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     8281 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     8341 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     8401 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     8461 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     8521 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     8581 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     8641 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     8701 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     8761 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     8821 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     8881 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     8941 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     9001 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     9061 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     9121 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     9181 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     9241 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     9301 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     9361 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     9421 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     9481 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     9541 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     9601 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     9661 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     9721 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     9781 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     9841 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     9901 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     9961 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
    10021 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
    10081 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
    10141 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
    10201 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
    10261 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
    10321 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
    10381 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
    10441 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
    10501 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
    10561 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
    10621 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
    10681 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaaga gagaaaggcg
    10741 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
    10801 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
    10861 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
    10921 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
    10981 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
    11041 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
    11101 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
    11161 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
    11221 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
    11281 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
    11341 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
    11401 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc
    11461 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
    11521 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
    11581 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
    11641 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtg
//
