LOCUS       sequence-174094-       10109 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of human codon-optimized dCas9-DNMT3A fusion with 
            T2A-EGFP for targeted DNA methylation in mammalian cells; cloning 
            backbone for sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 10109)
  AUTHORS   Vojta A, Dobrinic P, Tadic V, Bockor L, Korac P, Julg B, Klasic M, 
            Zoldos V
  TITLE     Repurposing the CRISPR-Cas9 system for targeted DNA methylation.
  JOURNAL   Nucleic Acids Res. 2016 Mar 11. pii: gkw159.
  PUBMED    26969735
REFERENCE   2  (bases 1 to 10109)
  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: "Nucleic 
            Acids Res. 2016 Mar 11. pii: gkw159."
COMMENT     Sequence Label: pdCas9-DNMT3A-EGFP
FEATURES             Location/Qualifiers
     source          1..10109
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        119..404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     misc_RNA        complement(501..576)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(595..843)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(653..672)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(823..843)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(905..1493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(985..1004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1664..2455,2456..2524))
                     /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     2287..2306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2525..2629)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2697..2715
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2753..2775)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2875..2894
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2911..3366)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3048..3069)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3260..3279
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3441..3581
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3452..3581
                     /label=AAV2 ITR
     polyA_signal    3590..3797
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3786..3803
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(3836..4546,4547..4549))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(3836..4546,4547..4549))
                     /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(3878..3899)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4227..4246
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(4550..4603)
                     /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             complement(4631..5563)
                     /codon_start=1
                     /product="catalytic domain of DNA methyltransferase 3A"
                     /label=DNMT3A catalytic domain
                     /translation="PSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATG
                     LLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVI
                     GGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVS
                     DKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGR
                     IAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSR
                     LARQRLLGRSWSVPVIRHLFAPLKEYFACV"
     CDS             complement(5591..5638)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(join(5639..7222,7223..7225,7226..9712,9713..9715,
                     9716..9739))
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(5639..9739)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(9764..9784)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(9791..9856)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      9856..9865
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          9877..10104
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
ORIGIN
        1 cccccccccc cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg
       61 ggggtgggga ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca
      121 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
      181 tgtactgggc acaatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
      241 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
      301 attgacgtca atggaaagtc cctattggcg ttactattga cgtcaatggg cgggggtcgt
      361 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgggtacc tctagagcca
      421 tttgtctgca gaattggcgc acgcgctaaa aacggactag ccttatttta acttgctatt
      481 tctagctcta aaacaaaaaa gcaccgactc ggtgccactt tttcaagttg ataacggact
      541 agccttattt taacttgcta tttctagctc taaaacaggt cttctcgaag acccggtgtt
      601 tcgtcctttc cacaagatat ataaagccaa gaaatcgaaa tactttcaag ttacggtaag
      661 catatgatag tccattttaa aacataattt taaaactgca aactacccaa gaaattatta
      721 ctttctacgt cacgtatttt gtactaatat ctttgtgttt acagtcaaat taattccaat
      781 tatctctcta acagccttgt atcgtatatg caaatatgaa ggaatcatgg gaaataggcc
      841 ctcacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      901 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      961 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1021 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1081 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1141 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1201 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1261 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1321 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1381 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1441 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1501 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1561 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1621 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     1681 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     1741 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     1801 cgcggcttcc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     1861 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     1921 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     1981 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     2041 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     2101 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     2161 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     2221 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     2281 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     2341 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     2401 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     2461 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     2521 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     2581 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     2641 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     2701 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     2761 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     2821 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     2881 cagagcagat tgtactgaga gtgcaccata aaattgtaaa cgttaatatt ttgttaaaat
     2941 tcgcgttaaa tttttgttaa atcagctcat tttttaacca atagaccgaa atcggcaaaa
     3001 tcccttataa atcaaaagaa tagcccgaga tagagttgag tgttgttcca gtttggaaca
     3061 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3121 gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta
     3181 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3241 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctaag gcgctggcaa
     3301 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3361 gcgcgtacta tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat gcgtaaggag
     3421 aaaataccgc atcaggcgcc cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     3481 ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
     3541 gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgct ccccagcatg
     3601 cctgctattc tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat
     3661 agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt
     3721 gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg
     3781 gcaactagaa ggcacagtcg aggctgatca gcgagctcta gttagaattg aattccttgt
     3841 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     3901 gatcgcgctt ctcgttgggg tctttgctca gggcggactg ggtgctcagg tagtggttgt
     3961 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
     4021 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct
     4081 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
     4141 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     4201 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     4261 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     4321 ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc
     4381 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
     4441 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     4501 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcact gggccaggat
     4561 tctcctcgac gtcaccgcat gttagcagac ttcctctgcc ctctccactg ccgaattcgg
     4621 gccggccgga cacacacgca aaatactcct tcagcggagc gaagaggtgg cggatgactg
     4681 gcacgctcca tgaccggccc agcagtctct gcctcgccaa gcggctcatg ttggagacgt
     4741 cagtatagtg gactgggaaa ccaaataccc tttccatttc agtgcaccat aagatgtcct
     4801 ctttctcatt catgaacaca ggaaaatgct ggtctttgcc ctgctttatg gagtttgacc
     4861 tcgtagtaat ggtcctcact ttgctgaact tggctatcct gccatgctcc agacactcct
     4921 gcagctccag cttatcattc acagtggatg ccaacggcct gttcataccg ggaaggttac
     4981 cccagaagta gcgggccctg tgtgcagctg acacttcttt ggcatcaatc atcacagggt
     5041 tggactcgag aaatcgcgag atgtccctct tgtcactaac gcccatggcc accacattct
     5101 caaagagcca gaagaagggg cgatcatctc cctccttggg ccgcgcatca tgcaggaggc
     5161 ggtagaactc aaagaagagc cggccagtgc cctcgtagag gcccttgcga gcagggttga
     5221 cgatggagag gtcattgcag ggactgcccc caatcaccag atcgaatggg ccccactcct
     5281 ggatatgctt ctgtgtgacg ctgcggacgt ccccgacgta catgatcttc ccctggtgcc
     5341 gcaccatgcc caccgtgatg gagtcctcac acacctccga ggcaatgtag cggtccacct
     5401 gaatgcccaa gtccttcagc accaggagcc ctgtagcgat tccatcaaag agagacagca
     5461 cccggatggg cttcctcttc tcagctggga caggtgggta aacctttgga gggtcaaatt
     5521 cctggtcgtg gttattagcg aagaacatct ggagccggga gggggatccg ctcccgcctc
     5581 cgccctcgag ctttttcttt ttggcctgtc cggccttttt cgtggccgcc ggccttttgt
     5641 cgcctcccag ctgagacagg tcgatccgtg tctcgtacag gccggtgatg ctctggtgga
     5701 tcagggtggc gtccagcacc tctttggtgc tggtgtacct cttccggtcg atggtggtgt
     5761 caaagtactt gaaggcggca ggggctccca gattggtcag ggtaaacagg tggatgatat
     5821 tctcggcctg ctctctgatg ggcttatccc ggtgcttgtt gtaggcggac agcactttgt
     5881 ccagattagc gtcggccagg atcactctct tggagaactc gctgatctgc tcgatgatct
     5941 cgtccaggta gtgcttgtgc tgttccacaa acagctgttt ctgctcatta tcctcggggg
     6001 agcccttcag cttctcatag tggctggcca ggtacaggaa gttcacatat ttggagggca
     6061 gggccagttc gtttcccttc tgcagttcgc cggcagaggc cagcattctc ttccggccgt
     6121 tttccagctc gaacagggag tacttaggca gcttgatgat caggtccttt ttcacttctt
     6181 tgtagccctt ggcttccaga aagtcgatgg gattcttctc gaagctgctt ctttccatga
     6241 tggtgatccc cagcagctct ttcacactct tcagtttctt ggacttgccc ttttccactt
     6301 tggccaccac cagcacagaa taggccacgg tggggctgtc gaagccgccg tacttcttag
     6361 ggtcccagtc cttctttctg gcgatcagct tatcgctgtt cctcttgggc aggatagact
     6421 ctttgctgaa gccgcctgtc tgcacctcgg tctttttcac gatattcact tggggcatgc
     6481 tcagcacttt ccgcacggtg gcaaaatccc ggcccttatc ccacacgatc tccccggttt
     6541 cgccgtttgt ctcgatcaga ggccgcttcc ggatctcgcc gttggccagg gtaatctcgg
     6601 tcttgaaaaa gttcatgatg ttgctgtaga agaagtactt ggcggtagcc ttgccgattt
     6661 cctgctcgct cttggcgatc atcttccgca cgtcgtacac cttgtagtcg ccgtacacga
     6721 actcgctttc cagcttaggg tactttttga tcagggcggt tcccacgacg gcgttcaggt
     6781 aggcgtcgtg ggcgtggtgg tagttgttga tctcgcgcac tttgtaaaac tggaaatcct
     6841 tccggaaatc ggacaccagc ttggacttca gggtgatcac tttcacttcc cggatcagct
     6901 tgtcattctc gtcgtactta gtgttcatcc gggagtccag gatctgtgcc acgtgctttg
     6961 tgatctgccg ggtttccacc agctgtctct tgatgaagcc ggccttatcc agttcgctca
     7021 ggccgcctct ctcggccttg gtcagattgt cgaactttct ctgggtaatc agcttggcgt
     7081 tcagcagctg ccgccagtag ttcttcatct tcttcacgac ctcttcggag ggcacgttgt
     7141 cgctcttgcc ccggttcttg tcgcttctgg tcagcacctt gttgtcgatg gagtcgtcct
     7201 tcagaaagct ctgaggcacg atagcgtcca catcgtagtc ggacagccgg ttgatgtcca
     7261 gttcctggtc cacgtacata tcccgcccat tctgcaggta gtacaggtac agcttctcgt
     7321 tctgcagctg ggtgttttcc acggggtgtt ctttcaggat ctggctgccc agctctttga
     7381 tgccctcttc gatccgcttc attctctcgc ggctgttctt ctgtcccttc tgggtggtct
     7441 ggttctctct ggccatttcg atcacgatgt tctcgggctt gtgccggccc atcactttca
     7501 cgagctcgtc caccaccttc actgtctgca ggatgccctt cttaatggcg gggctgccgg
     7561 ccagattggc aatgtgctcg tgcaggctat cgccctggcc ggacacctgg gctttctgga
     7621 tgtcctcttt aaaggtcagg ctgtcgtcgt ggatcagctg catgaagttt ctgttggcga
     7681 agccgtcgga cttcaggaaa tccaggattg tcttgccgga ctgcttgtcc cggatgccgt
     7741 tgatcagctt ccggctcagc ctgccccagc cggtgtatct ccgccgcttc agctgcttca
     7801 tcactttgtc gtcgaacagg tgggcatagg ttttcagccg ttcctcgatc atctctctgt
     7861 cctcaaacag tgtcagggtc agcacgatat cttccagaat gtcctcgttt tcctcattgt
     7921 ccaggaagtc cttgtccttg ataattttca gcagatcgtg gtatgtgccc agggaggcgt
     7981 tgaaccgatc ttccacgccg gagatttcca cggagtcgaa gcactcgatt ttcttgaagt
     8041 agtcctcttt cagctgcttc acggtcactt tccggttggt cttgaacagc aggtccacga
     8101 tggccttttt ctgctcgccg ctcaggaagg cgggctttct cattccctcg gtcacgtatt
     8161 tcactttggt cagctcgtta tacacggtga agtactcgta cagcaggctg tgcttgggca
     8221 gcaccttctc gttgggcagg ttcttatcga agttggtcat ccgctcgatg aagctctggg
     8281 cggaagcgcc cttgtccacc acttcctcga agttccaggg ggtgatggtt tcctcgctct
     8341 ttctggtcat ccaggcgaat ctgctgtttc ccctggccag agggcccacg tagtagggga
     8401 tgcggaaggt caggatcttc tcgatctttt cccggttgtc cttcaggaat gggtaaaaat
     8461 cttcctgccg ccgcagaatg gcgtgcagct ctcccaggtg gatctggtgg gggatgctgc
     8521 cgttgtcgaa ggtccgctgc ttccgcagca ggtcctctct gttcagcttc acgagcagtt
     8581 cctcggtgcc gtccatcttt tccaggatgg gcttgatgaa cttgtagaac tcttcctggc
     8641 tggctccgcc gtcaatgtag ccggcgtagc cgttcttgct ctggtcgaag aaaatctctt
     8701 tgtacttctc aggcagctgc tgccgcacga gagctttcag cagggtcagg tcctggtggt
     8761 gctcgtcgta tctcttgatc atagaggcgc tcaggggggc cttggtgatc tcggtgttca
     8821 ctctcaggat gtcgctcagc aggatggcgt cggacaggtt cttggcggcc agaaacaggt
     8881 cggcgtactg gtcgccgatc tgggccagca ggttgtccag gtcgtcgtcg taggtgtcct
     8941 tgctcagctg cagtttggca tcctcggcca ggtcgaagtt gctcttgaag ttgggggtca
     9001 ggcccaggct cagggcaatc aggttgccga acaggccatt cttcttctcg ccgggcagct
     9061 gggcgatcag attttccagc cgtctgctct tgctcagtct ggcagacagg atggccttgg
     9121 cgtccacgcc gctggcgttg atggggtttt cctcgaacag ctggttgtag gtctgcacca
     9181 gctggatgaa cagcttgtcc acgtcgctgt tgtcggggtt caggtcgccc tcgatcagga
     9241 agtggccccg gaacttgatc atgtgggcca gggccagata gatcagccgc aggtcggcct
     9301 tgtcggtgct gtccaccagt ttctttctca ggtggtagat ggtggggtac ttctcgtggt
     9361 aggccacctc gtccacgatg ttgccgaaga tggggtgccg ctcgtgcttc ttatcctctt
     9421 ccaccaggaa ggactcttcc agtctgtgga agaagctgtc gtccaccttg gccatctcgt
     9481 tgctgaagat ctcttgcaga tagcagatcc ggttcttccg tctggtgtat cttcttctgg
     9541 cggttctctt cagccgggtg gcctcggctg tttcgccgct gtcgaacagc agggctccga
     9601 tcaggttctt cttgatgctg tgccggtcgg tgttgcccag caccttgaat ttcttgctgg
     9661 gcaccttgta ctcgtcggtg atcacggccc agcccacaga gttggtgccg atggccaggc
     9721 cgatgctgta cttcttgtcg gctgctggga ctccgtggat accgaccttc cgcttcttct
     9781 ttggggccat cttatcgtca tcgtctttgt aatcaatatc atgatccttg tagtctccgt
     9841 cgtggtcctt atagtccatg gtggcaccgg tccaacctga aaaaaagtga tttcaggcag
     9901 gtgctccagg taattaaaca ttaatacccc accaaccaac catcccttaa acccttacct
     9961 cttgctcagc taattacagc ccggaggaga agggccgtcc cgcccgctca cctgtgggag
    10021 taacgcggtc agtcagagcc ggggcgggcg gcgcgaggcg gcggcggagc ggggcacggg
    10081 gcgaaggcag cgcgcagcga ctcccgccc
//
