LOCUS       sequence-457017-        9760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR targeting of human EHMT1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9760)
  AUTHORS   Biligiri KK, Sharma NR, Mohanty A, Sarkar DP, Vemula PK, Rampalli S.
  TITLE     A cytoplasmic form of EHMT1N methylates viral proteins to enable 
            inclusion body maturation and efficient viral replication
  JOURNAL   PLOS Biology 22(11): e3002871.
REFERENCE   2  (bases 1 to 9760)
  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: "PLOS 
            Biology 22(11): e3002871."
COMMENT     Sequence Label: EHMT1_exon 3_gRNA
FEATURES             Location/Qualifiers
     source          1..9760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             827..874
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             890..943
                     /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(944..946,947..1657)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(944..946,947..1657)
                     /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(1247..1266)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1594..1615
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1685..1702)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1691..1898
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1907..2047
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1907..2036
                     /label=AAV2 ITR
     rep_origin      2122..2577
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2209..2228)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2419..2440
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2594..2613)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2713..2735
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2773..2791)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2859..2963
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2964..3032,3033..3824)
                     /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(3182..3201)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3995..4583
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4484..4503
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4645..4893
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        4915..4990
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        5081..5329
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        5356..5431
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        5521..5806
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          6121..6348
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      6360..6369
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6369..6434
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6441..6461
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(6486..9760,1..826)
                     /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             join(6486..9760,1..826)
                     /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"
ORIGIN
        1 tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa
       61 gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg
      121 cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa
      181 gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag
      241 cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga
      301 cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat
      361 gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt
      421 gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga
      481 gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat
      541 cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc
      601 ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt
      661 taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg
      721 gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac
      781 cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc
      841 cacgaaaaag gccggccagg caaaaaagaa aaaggaattc ggcagtggag agggcagagg
      901 aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccagtgagca agggcgagga
      961 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1021 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1081 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     1141 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1201 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1261 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1321 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1381 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     1441 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1501 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     1561 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1621 cgccgggatc actctcggca tggacgagct gtacaaggaa ttctaactag agctcgctga
     1681 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1741 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1801 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1861 ggggaggatt gggaagagaa tagcaggcat gctggggagc ggccgcagga acccctagtg
     1921 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     1981 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     2041 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2101 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2161 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     2221 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2281 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     2341 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     2401 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     2461 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     2521 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     2581 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     2641 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     2701 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     2761 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     2821 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2881 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2941 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3001 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3061 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3121 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3181 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3241 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3301 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3361 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3421 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3481 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3541 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3601 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3661 ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca gatggtaagc
     3721 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3781 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3841 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3901 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3961 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4021 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4081 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4141 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4201 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4261 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4321 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4381 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4441 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4501 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4561 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4621 tttgctggcc ttttgctcac atgtgagggc ctatttccca tgattccttc atatttgcat
     4681 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
     4741 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
     4801 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
     4861 gctttatata tcttgtggaa aggacgaaac accgcgccga cgtcaaggtc cacagtttta
     4921 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
     4981 gagtcggtgc ttttttgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
     5041 tttagcgcgt gcgccaattc tgcagacaaa tggctctaga gagggcctat ttcccatgat
     5101 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac
     5161 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
     5221 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa
     5281 gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg gagacctatt
     5341 cgccttaagg tctcggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     5401 tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt ttagagctag aaatagcaag
     5461 ttaaaataag actagtccgt ttttagcgcg tgcgccaatt ctgcagacaa atggggtacc
     5521 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     5581 gacgtcaata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     5641 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     5701 cgtcaatgac ggtaaatggc ccgcctggca ttgtgcccag tacatgacct tatgggactt
     5761 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
     5821 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
     5881 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggg cgccccggcg
     5941 gggggggggg cggggggggg gggggggggg gggggggggg gggggggggg cgaggggcgg
     6001 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
     6061 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
     6121 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
     6181 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
     6241 ccgggctgta attagctgag caagaggtaa gggtttaagg gatggttggt tggtggggta
     6301 ttaatgttta attacctgga gcacctgcct gaaatcactt tttttcaggt tggaccggtg
     6361 ccaccatgga ctataaggac cacgacggag actacaagga tcatgatatt gattacaaag
     6421 acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac ggagtcccag
     6481 cagccgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg
     6541 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
     6601 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
     6661 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
     6721 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
     6781 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
     6841 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
     6901 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
     6961 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
     7021 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
     7081 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
     7141 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa
     7201 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
     7261 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
     7321 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
     7381 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct
     7441 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc
     7501 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg
     7561 ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc
     7621 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc
     7681 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc
     7741 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga
     7801 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca
     7861 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg
     7921 aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata
     7981 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg
     8041 tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc
     8101 tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga
     8161 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa
     8221 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa
     8281 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg
     8341 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg
     8401 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg
     8461 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc
     8521 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg
     8581 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc
     8641 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga
     8701 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg
     8761 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga
     8821 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc
     8881 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc
     8941 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc
     9001 atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa
     9061 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga
     9121 agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc
     9181 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac
     9241 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga
     9301 acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt
     9361 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca
     9421 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa
     9481 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga
     9541 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca
     9601 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc
     9661 ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt
     9721 ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa
//
