LOCUS       sequence-150190-       14332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dead Cas9 (dCas9) fused to DNMT3A catalytic domain (CD) 
            under the control of the CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14332)
  AUTHORS   McDonald JI, Celik H, Rois LE, Fishberger G, Fowler T, Rees R, 
            Kramer A, Martens A, Edwards JR, Challen GA
  TITLE     Reprogrammable CRISPR/Cas9-based system for inducing site-specific 
            DNA methylation.
  JOURNAL   Biol Open. 2016 May 11. pii: bio.019067. doi: 10.1242/bio.019067.
  PUBMED    27170255
REFERENCE   2  (bases 1 to 14332)
  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: "Biol Open. 
            2016 May 11. pii: bio.019067. doi: 10.1242/bio.019067."
COMMENT     Sequence Label: pCMV-dCas9-D3A
FEATURES             Location/Qualifiers
     source          1..14332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2611..3821
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3658..3677
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      3856..3865
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3862..3888,3889..3891,3892..6375,6376..6381,6382..6447,
                     6448..6450,6451..7965)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEIAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             3862..7965
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEIAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7978..7998
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8008..8031
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8050..8982
                     /codon_start=1
                     /product="catalytic domain of DNA methyltransferase 3A"
                     /label=DNMT3A catalytic domain
                     /translation="PSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATG
                     LLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVI
                     GGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVS
                     DKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGR
                     IAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSR
                     LARQRLLGRSWSVPVIRHLFAPLKEYFACV"
     misc_feature    9024..9612
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9077..9097)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9495..9506)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9615..9631)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9616..9632)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10137..10317
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10343..10360)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10349..10573
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10619..11047
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10706..10725)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10916..10937
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11056..11076)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11061..11390
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11241..11376
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11303..11322
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11438..11485
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11504..11878
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    12008..12129
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12045..12064)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12099..12118
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12178..12194)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12178..12194)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12191..12213)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12202..12218
                     /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)."
     promoter        complement(join(12226..12232,12233..12250,12251..12256))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12271..12292
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12409..12426)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12580..13168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12660..12679)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13339..14130,14131..14199))
                     /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     13962..13981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14200..14304)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta agggtgcagc ggcctccgcg ccgggttttg gcgcctcccg
     2641 cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc aggagcgttc
     2701 ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag
     2761 aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg
     2821 ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg
     2881 agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac
     2941 agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc
     3001 gtcacttggt gagttgcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct
     3061 cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag
     3121 gttgccctga actgggggtt ggggggagcg cacaaaatgg cggctgttcc cgagtcttga
     3181 atggaagacg cttgtaaggc gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg
     3241 gcggcaagaa cccaaggtct tgaggccttc gctaatgcgg gaaagctctt attcgggtga
     3301 gatgggctgg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc
     3361 gggtttgtcg tctggttgcg ggggcggcag ttatgcggtg ccgttgggca gtgcacccgt
     3421 acctttggga gcgcgcgcct cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg
     3481 cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg
     3541 gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg
     3601 gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt aagtagctga
     3661 agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa gttttttagg
     3721 caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag actagtaaat
     3781 tgtccgctaa attctggccg tttttggctt ttttgttaga cgaagcttgg gctgcaggtc
     3841 gactctagag gatccgccac catggacaag aagtactcca ttgggctcgc tatcggcaca
     3901 aacagcgtcg gctgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa
     3961 gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggcgc cctcctgttc
     4021 gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc
     4081 cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat agctaaggtg
     4141 gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagcac
     4201 gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca
     4261 accatatatc atctgaggaa gaagcttgta gacagtactg ataaggctga cttgcggttg
     4321 atctatctcg cgctggcgca tatgatcaaa tttcggggac acttcctcat cgagggggac
     4381 ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat
     4441 cagcttttcg aagagaaccc gatcaacgca tccggagttg acgccaaagc aatcctgagc
     4501 gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag
     4561 aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa
     4621 tctaacttcg acctggccga agatgccaag cttcaactga gcaaagacac ctacgatgat
     4681 gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca
     4741 aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc
     4801 aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact
     4861 ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat
     4921 cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga ggaattttac
     4981 aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt
     5041 aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag
     5101 attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg
     5161 aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc
     5221 cccctcgccc ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc
     5281 actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa
     5341 aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg
     5401 ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg
     5461 atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc
     5521 aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa aaagattgaa
     5581 tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg
     5641 tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag
     5701 gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa
     5761 gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagagg
     5821 cgccgatata caggatgggg gcggctgtca agaaaactga tcaatgggat ccgagacaag
     5881 cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa ccggaacttc
     5941 atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa agcacaagtt
     6001 tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag cccagctatc
     6061 aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg
     6121 cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac ccagaaggga
     6181 cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga actggggtcc
     6241 caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa gctctacctg
     6301 tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat caatcggctc
     6361 tccgactacg acgtggatgc tatcgtgccc cagtcttttc tcaaagatga ttctattgat
     6421 aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt cccctcagaa
     6481 gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa actgatcaca
     6541 caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga gttggataaa
     6601 gccggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca cgtggcccaa
     6661 attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat tcgagaggtg
     6721 aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt tcagttttat
     6781 aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa tgcagtggta
     6841 ggcactgcac ttatcaaaaa atatcccaag cttgaatctg aatttgttta cggagactat
     6901 aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg caaggccacc
     6961 gctaagtact tcttttacag caatattatg aattttttca agaccgagat tacactggcc
     7021 aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg agaaatcgtg
     7081 tgggacaagg gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc gcaggtgaac
     7141 atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat cctcccgaaa
     7201 aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa atacggcgga
     7261 ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag
     7321 tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caatcatgga gcgatcaagc
     7381 ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat ataaagaggt caaaaaagac
     7441 ctcatcatta agcttcccaa gtactctctc tttgagcttg aaaacggccg gaaacgaatg
     7501 ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc taaatacgtt
     7561 aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga agataatgag
     7621 cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat cgagcaaata
     7681 agcgaattct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt gctttctgct
     7741 tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat ccacttgttt
     7801 actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac catagacaga
     7861 aagcggtaca cctctacgaa ggaggtcctg gacgccacac tgattcatca gtcaattacg
     7921 gggctctatg aaacaagaat cgacctctct cagctcggtg gagacagcag ggctgacccc
     7981 aagaagaaga ggaaggtggg agctagcgac tacaaagacg atgacgacaa gggaagccga
     8041 gaggactggc cctcccggct ccagatgttc ttcgctaata accacgacca ggaatttgac
     8101 cctccaaagg tttacccacc tgtcccagct gagaagagga agcccatccg ggtgctgtct
     8161 ctctttgatg gaatcgctac agggctcctg gtgctgaagg acttgggcat tcaggtggac
     8221 cgctacattg cctcggaggt gtgtgaggac tccatcacgg tgggcatggt gcggcaccag
     8281 gggaagatca tgtacgtcgg ggacgtccgc agcgtcacac agaagcatat ccaggagtgg
     8341 ggcccattcg atctggtgat tgggggcagt ccctgcaatg acctctccat cgtcaaccct
     8401 gctcgcaagg gcctctacga gggcactggc cggctcttct ttgagttcta ccgcctcctg
     8461 catgatgcgc ggcccaagga gggagatgat cgccccttct tctggctctt tgagaatgtg
     8521 gtggccatgg gcgttagtga caagagggac atctcgcgat ttctcgagtc caaccctgtg
     8581 atgattgatg ccaaagaagt gtcagctgca cacagggccc gctacttctg gggtaacctt
     8641 cccggtatga acaggccgtt ggcatccact gtgaatgata agctggagct gcaggagtgt
     8701 ctggagcatg gcaggatagc caagttcagc aaagtgagga ccattactac gaggtcaaac
     8761 tccataaagc agggcaaaga ccagcatttt cctgtcttca tgaatgagaa agaggacatc
     8821 ttatggtgca ctgaaatgga aagggtattt ggtttcccag tccactatac tgacgtctcc
     8881 aacatgagcc gcttggcgag gcagagactg ctgggccggt catggagcgt gccagtcatc
     8941 cgccacctct tcgctccgct gaaggagtat tttgcgtgtg tgtaaaccgg tctagctaag
     9001 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     9061 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     9121 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     9181 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     9241 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     9301 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     9361 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     9421 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     9481 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     9541 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     9601 ccgcctcccc gcatcgatac cgtcgacctc gagacctaga aaaacatgga gcaatcacaa
     9661 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg
     9721 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg
     9781 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     9841 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc
     9901 agaactacac accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc
     9961 tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt
    10021 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt
    10081 ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt
    10141 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
    10201 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
    10261 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg
    10321 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
    10381 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
    10441 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
    10501 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
    10561 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
    10621 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
    10681 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
    10741 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
    10801 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
    10861 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
    10921 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
    10981 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
    11041 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
    11101 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
    11161 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
    11221 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
    11281 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
    11341 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
    11401 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
    11461 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
    11521 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
    11581 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
    11641 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
    11701 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
    11761 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
    11821 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
    11881 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
    11941 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
    12001 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
    12061 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
    12121 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
    12181 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
    12241 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
    12301 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
    12361 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
    12421 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
    12481 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
    12541 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
    12601 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
    12661 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
    12721 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
    12781 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
    12841 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
    12901 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
    12961 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
    13021 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
    13081 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
    13141 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
    13201 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
    13261 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
    13321 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
    13381 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
    13441 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
    13501 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
    13561 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
    13621 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
    13681 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
    13741 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
    13801 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
    13861 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
    13921 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
    13981 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
    14041 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
    14101 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
    14161 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
    14221 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
    14281 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ac
//
