LOCUS       sequence-254626-       16074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct to express dCas9 fused with Dnmt3a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16074)
  AUTHORS   Liu XS, Wu H, Ji X, Stelzer Y, Wu X, Czauderna S, Shu J, Dadon D, 
            Young RA, Jaenisch R
  TITLE     Editing DNA Methylation in the Mammalian Genome.
  JOURNAL   Cell. 2016 Sep 22;167(1):233-247.e17. doi: 
            10.1016/j.cell.2016.08.056.
  PUBMED    27662091
REFERENCE   2  (bases 1 to 16074)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2016.08"; journalName: "Cell"; date: "2016-09-22- 
            22"; volume: "167"; issue: "1"; pages: "233-247.e17"
COMMENT     Sequence Label: Fuw-dCas9-Dnmt3a
FEATURES             Location/Qualifiers
     source          1..16074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     209..230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(349..369)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        393..603
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        415..611
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      462..597
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     524..543
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        659..706
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             725..1099
                     /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    1229..1350
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1266..1285)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1320..1339
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1399..1415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1399..1415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1412..1434)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1423..1439
                     /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(1447..1453,1454..1471,1472..1477))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1492..1513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1630..1647)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1801..2389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1881..1900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2560..3351,3352..3420))
                     /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     3183..3202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3421..3525)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3600..3619)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3791..4170
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4172..4370
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4325..4345
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4388..4568
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4615..4740
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5233..5466
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5651..5695
                     /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    5993..6110
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6175..7386
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     7223..7242
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(7437..7463,7464..7466,7467..9953,9954..9956,
                     9957..11540)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             11553..11573
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11577..11597
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11601..11621
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11628..14366
                     /codon_start=1
                     /product="DNA methyltransferase 3A"
                     /label=DNMT3A
                     /translation="MPAMPSSGPGDTSSSAAEREEDRKDGEEQEEPRGKEERQEPSTTA
                     RKVGRPGRKRKHPPVESGDTPKDPAVISKSPSMAQDSGASELLPNGDLEKRSEPQPEEG
                     SPAGGQKGGAPAEGEGAAETLPEASRAVENGCCTPKEGRGAPAEAGKEQKETNIESMKM
                     EGSRGRLRGGLGWESSLRQRPMPRLTFQAGDPYYISKRKRDEWLARWKREAEKKAKVIA
                     GMNAVEENQGPGESQKVEEASPPAVQQPTDPASPTVATTPEPVGSDAGDKNATKAGDDE
                     PEYEDGRGFGIGELVWGKLRGFSWWPGRIVSWWMTGRSRAAEGTRWVMWFGDGKFSVVC
                     VEKLMPLSSFCSAFHQATYNKQPMYRKAIYEVLQVASSRAGKLFPVCHDSDESDTAKAV
                     EVQNKPMIEWALGGFQPSGPKGLEPPEEEKNPYKEVYTDMWVEPEAAAYAPPPPAKKPR
                     KSTAEKPKVKEIIDERTRERLVYEVRQKCRNIEDICISCGSLNVTLEHPLFVGGMCQNC
                     KNCFLECAYQYDDDGYQSYCTICCGGREVLMCGNNNCCRCFCVECVDLLVGPGAAQAAI
                     KEDPWNCYMCGHKGTYGLLRRREDWPSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIR
                     VLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKH
                     IQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFW
                     LFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDK
                     LELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFP
                     VHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV"
     misc_feature    14391..14979
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(14444..14464)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(14862..14873)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(14982..14998)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14983..14999)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             15504..15684
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(15710..15727)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    15716..15940
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(15986..16074,1..340)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(join(16073..16074,1..18))
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
       61 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
      121 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
      181 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
      241 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
      301 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
      361 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
      421 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
      481 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
      541 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
      601 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
      661 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
      721 aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc
      781 ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
      841 cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc
      901 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
      961 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
     1021 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
     1081 ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga
     1141 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     1201 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     1261 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     1321 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     1381 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1441 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1501 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1561 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1621 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1681 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1741 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1801 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1861 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1921 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1981 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2041 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2101 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2161 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2221 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2281 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2341 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2401 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2461 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2521 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2581 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2641 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2701 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2761 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2821 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2881 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2941 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3001 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3061 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3121 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3181 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3241 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3301 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3361 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3421 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3481 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3541 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     3601 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     3661 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     3721 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     3781 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     3841 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     3901 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     3961 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     4021 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     4081 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     4141 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     4201 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     4261 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     4321 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc
     4381 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     4441 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     4501 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     4561 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
     4621 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
     4681 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
     4741 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
     4801 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
     4861 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     4921 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     4981 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     5041 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg
     5101 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     5161 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     5221 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     5281 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     5341 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     5401 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     5461 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     5521 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
     5581 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     5641 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     5701 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     5761 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     5821 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
     5881 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
     5941 atcggcactg cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa
     6001 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     6061 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     6121 acagggacag cagagatcca gtttggttaa ttaacccgtg tcggctccag atctggcctc
     6181 cgcgccgggt tttggcgcct cccgcgggcg cccccctcct cacggcgagc gctgccacgt
     6241 cagacgaagg gcgcagcgag cgtcctgatc cttccgcccg gacgctcagg acagcggccc
     6301 gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg
     6361 acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt
     6421 agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat
     6481 ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt
     6541 tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctggccgg
     6601 ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg
     6661 ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg agcgcagcaa
     6721 aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt
     6781 tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagg ccttcgctaa
     6841 tgcgggaaag ctcttattcg ggtgagatgg gctggggcac catctgggga ccctgacgtg
     6901 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     6961 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     7021 tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg tgtgcggtag
     7081 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     7141 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     7201 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     7261 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     7321 aattttcagt gttagactag taaattgtcc gctaaattct ggccgttttt ggcttttttg
     7381 ttagacgaag cttgggctgc aggtcgactc tagaagttaa cctcgagggc gcgcccatgg
     7441 acaagaagta ttctatcgga ctggccatcg ggactaatag cgtcgggtgg gccgtgatca
     7501 ctgacgagta caaggtgccc tctaagaagt tcaaggtgct cgggaacacc gaccggcatt
     7561 ccatcaagaa aaatctgatc ggagctctcc tctttgattc aggggagacc gctgaagcaa
     7621 cccgcctcaa gcggactgct agacggcggt acaccaggag gaagaaccgg atttgttacc
     7681 ttcaagagat attctccaac gaaatggcaa aggtcgacga cagcttcttc cataggctgg
     7741 aagaatcatt cctcgtggaa gaggataaga agcatgaacg gcatcccatc ttcggtaata
     7801 tcgtcgacga ggtggcctat cacgagaaat acccaaccat ctaccatctt cgcaaaaagc
     7861 tggtggactc aaccgacaag gcagacctcc ggcttatcta cctggccctg gcccacatga
     7921 tcaagttcag aggccacttc ctgatcgagg gcgacctcaa tcctgacaat agcgatgtgg
     7981 ataaactgtt catccagctg gtgcagactt acaaccagct ctttgaagag aaccccatca
     8041 atgcaagcgg agtcgatgcc aaggccattc tgtcagcccg gctgtcaaag agccgcagac
     8101 ttgagaatct tatcgctcag ctgccgggtg aaaagaaaaa tggactgttc gggaacctga
     8161 ttgctctttc acttgggctg actcccaatt tcaagtctaa tttcgacctg gcagaggatg
     8221 ccaagctgca actgtccaag gacacctatg atgacgatct cgacaacctc ctggcccaga
     8281 tcggtgacca atacgccgac cttttccttg ctgctaagaa tctttctgac gccatcctgc
     8341 tgtctgacat tctccgcgtg aacactgaaa tcaccaaggc ccctctttca gcttcaatga
     8401 ttaagcggta tgatgagcac caccaggacc tgaccctgct taaggcactc gtccggcagc
     8461 agcttccgga gaagtacaag gaaatcttct ttgaccagtc aaagaatgga tacgccggct
     8521 acatcgacgg aggtgcctcc caagaggaat tttataagtt tatcaaacct atccttgaga
     8581 agatggacgg caccgaagag ctcctcgtga aactgaatcg ggaggatctg ctgcggaagc
     8641 agcgcacttt cgacaatggg agcattcccc accagatcca tcttggggag cttcacgcca
     8701 tccttcggcg ccaagaggac ttctacccct ttcttaagga caacagggag aagattgaga
     8761 aaattctcac tttccgcatc ccctactacg tgggacccct cgccagagga aatagccggt
     8821 ttgcttggat gaccagaaag tcagaagaaa ctatcactcc ctggaacttc gaagaggtgg
     8881 tggacaaggg agccagcgct cagtcattca tcgaacggat gactaacttc gataagaacc
     8941 tccccaatga gaaggtcctg ccgaaacatt ccctgctcta cgagtacttt accgtgtaca
     9001 acgagctgac caaggtgaaa tatgtcaccg aagggatgag gaagcccgca ttcctgtcag
     9061 gcgaacaaaa gaaggcaatt gtggaccttc tgttcaagac caatagaaag gtgaccgtga
     9121 agcagctgaa ggaggactat ttcaagaaaa ttgaatgctt cgactctgtg gagattagcg
     9181 gggtcgaaga tcggttcaac gcaagcctgg gtacctacca tgatctgctt aagatcatca
     9241 aggacaagga ttttctggac aatgaggaga acgaggacat ccttgaggac attgtcctga
     9301 ctctcactct gttcgaggac cgggaaatga tcgaggagag gcttaagacc tacgcccatc
     9361 tgttcgacga taaagtgatg aagcaactta aacggagaag atataccgga tggggacgcc
     9421 ttagccgcaa actcatcaac ggaatccggg acaaacagag cggaaagacc attcttgatt
     9481 tccttaagag cgacggattc gctaatcgca acttcatgca acttatccat gatgattccc
     9541 tgacctttaa ggaggacatc cagaaggccc aagtgtctgg acaaggtgac tcactgcacg
     9601 agcatatcgc aaatctggct ggttcacccg ctattaagaa gggtattctc cagaccgtga
     9661 aagtcgtgga cgagctggtc aaggtgatgg gtcgccataa accagagaac attgtcatcg
     9721 agatggccag ggaaaaccag actacccaga agggacagaa gaacagcagg gagcggatga
     9781 aaagaattga ggaagggatt aaggagctcg ggtcacagat ccttaaagag cacccggtgg
     9841 aaaacaccca gcttcagaat gagaagctct atctgtacta ccttcaaaat ggacgcgata
     9901 tgtatgtgga ccaagagctt gatatcaaca ggctctcaga ctacgacgtg gacgccatcg
     9961 tccctcagag cttcctcaaa gacgactcaa ttgacaataa ggtgctgact cgctcagaca
    10021 agaaccgggg aaagtcagat aacgtgccct cagaggaagt cgtgaaaaag atgaagaact
    10081 attggcgcca gcttctgaac gcaaagctga tcactcagcg gaagttcgac aatctcacta
    10141 aggctgagag gggcggactg agcgaactgg acaaagcagg attcattaaa cggcaacttg
    10201 tggagactcg gcagattact aaacatgtcg cccaaatcct tgactcacgc atgaatacca
    10261 agtacgacga aaacgacaaa cttatccgcg aggtgaaggt gattaccctg aagtccaagc
    10321 tggtcagcga tttcagaaag gactttcaat tctacaaagt gcgggagatc aataactatc
    10381 atcatgctca tgacgcatat ctgaatgccg tggtgggaac cgccctgatc aagaagtacc
    10441 caaagctgga aagcgagttc gtgtacggag actacaaggt ctacgacgtg cgcaagatga
    10501 ttgccaaatc tgagcaggag atcggaaagg ccaccgcaaa gtacttcttc tacagcaaca
    10561 tcatgaattt cttcaagacc gaaatcaccc ttgcaaacgg tgagatccgg aagaggccgc
    10621 tcatcgagac taatggggag actggcgaaa tcgtgtggga caagggcaga gatttcgcta
    10681 ccgtgcgcaa agtgctttct atgcctcaag tgaacatcgt gaagaaaacc gaggtgcaaa
    10741 ccggaggctt ttctaaggaa tcaatcctcc ccaagcgcaa ctccgacaag ctcattgcaa
    10801 ggaagaagga ttgggaccct aagaagtacg gcggattcga ttcaccaact gtggcttatt
    10861 ctgtcctggt cgtggctaag gtggaaaaag gaaagtctaa gaagctcaag agcgtgaagg
    10921 aactgctggg tatcaccatt atggagcgca gctccttcga gaagaaccca attgactttc
    10981 tcgaagccaa aggttacaag gaagtcaaga aggaccttat catcaagctc ccaaagtata
    11041 gcctgttcga actggagaat gggcggaagc ggatgctcgc ctccgctggc gaacttcaga
    11101 agggtaatga gctggctctc ccctccaagt acgtgaattt cctctacctt gcaagccatt
    11161 acgagaagct gaaggggagc cccgaggaca acgagcaaaa gcaactgttt gtggagcagc
    11221 ataagcatta tctggacgag atcattgagc agatttccga gttttctaaa cgcgtcattc
    11281 tcgctgatgc caacctcgat aaagtcctta gcgcatacaa taagcacaga gacaaaccaa
    11341 ttcgggagca ggctgagaat atcatccacc tgttcaccct caccaatctt ggtgcccctg
    11401 ccgcattcaa gtacttcgac accaccatcg accggaaacg ctatacctcc accaaagaag
    11461 tgctggacgc caccctcatc caccagagca tcaccggact ttacgaaact cggattgacc
    11521 tctcacagct cggaggggat gagggagctg atccaaaaaa gaagagaaag gtagacccaa
    11581 agaaaaaacg aaaagtagat ccgaaaaaga agaggaaggt gggatccatg cccgccatgc
    11641 cctccagcgg ccccggggac accagcagct ctgctgcgga gcgggaggag gaccgaaagg
    11701 acggagagga gcaggaggag ccgcgtggca aggaggagcg ccaagagccc agcaccacgg
    11761 cacggaaggt ggggcggcct gggaggaagc gcaagcaccc cccggtggaa agcggtgaca
    11821 cgccaaagga ccctgcggtg atctccaagt ccccatccat ggcccaggac tcaggcgcct
    11881 cagagctatt acccaatggg gacttggaga agcggagtga gccccagcca gaggagggga
    11941 gccctgctgg ggggcagaag ggcggggccc cagcagaggg agagggtgca gctgagaccc
    12001 tgcctgaagc ctcaagagca gtggaaaatg gctgctgcac ccccaaggag ggccgaggag
    12061 cccctgcaga agcgggcaaa gaacagaagg agaccaacat cgaatccatg aaaatggagg
    12121 gctcccgggg ccggctgcgg ggtggcttgg gctgggagtc cagcctccgt cagcggccca
    12181 tgccgaggct caccttccag gcgggggacc cctactacat cagcaagcgc aagcgggacg
    12241 agtggctggc acgctggaaa agggaggctg agaagaaagc caaggtcatt gcaggaatga
    12301 atgctgtgga agaaaaccag gggcccgggg agtctcagaa ggtggaggag gccagccctc
    12361 ctgctgtgca gcagcccact gaccccgcat cccccactgt ggctaccacg cctgagcccg
    12421 tggggtccga tgctggggac aagaatgcca ccaaagcagg cgatgacgag ccagagtacg
    12481 aggacggccg gggctttggc attggggagc tggtgtgggg gaaactgcgg ggcttctcct
    12541 ggtggccagg ccgcattgtg tcttggtgga tgacgggccg gagccgagca gctgaaggca
    12601 cccgctgggt catgtggttc ggagacggca aattctcagt ggtgtgtgtt gagaagctga
    12661 tgccgctgag ctcgttttgc agtgcgttcc accaggccac gtacaacaag cagcccatgt
    12721 accgcaaagc catctacgag gtcctgcagg tggccagcag ccgcgcgggg aagctgttcc
    12781 cggtgtgcca cgacagcgat gagagtgaca ctgccaaggc cgtggaggtg cagaacaagc
    12841 ccatgattga atgggccctg gggggcttcc agccttctgg ccctaagggc ctggagccac
    12901 cagaagaaga gaagaatccc tacaaagaag tgtacacgga catgtgggtg gaacctgagg
    12961 cagctgccta cgcaccacct ccaccagcca aaaagccccg gaagagcaca gcggagaagc
    13021 ccaaggtcaa ggagattatt gatgagcgca caagagagcg gctggtgtac gaggtgcggc
    13081 agaagtgccg gaacattgag gacatctgca tctcctgtgg gagcctcaat gttaccctgg
    13141 aacaccccct cttcgttgga ggaatgtgcc aaaactgcaa gaactgcttt ctggagtgtg
    13201 cgtaccagta cgacgacgac ggctaccagt cctactgcac catctgctgt gggggccgtg
    13261 aggtgctcat gtgcggaaac aacaactgct gcaggtgctt ttgcgtggag tgtgtggacc
    13321 tcttggtggg gccgggggct gcccaggcag ccattaagga agacccctgg aactgctaca
    13381 tgtgcgggca caagggtacc tacgggctgc tgcggcggcg agaggactgg ccctcccggc
    13441 tccagatgtt cttcgctaat aaccacgacc aggaatttga ccctccaaag gtttacccac
    13501 ctgtcccagc tgagaagagg aagcccatcc gggtgctgtc tctctttgat ggaatcgcta
    13561 cagggctcct ggtgctgaag gacttgggca ttcaggtgga ccgctacatt gcctcggagg
    13621 tgtgtgagga ctccatcacg gtgggcatgg tgcggcacca ggggaagatc atgtacgtcg
    13681 gggacgtccg cagcgtcaca cagaagcata tccaggagtg gggcccattc gatctggtga
    13741 ttgggggcag tccctgcaat gacctctcca tcgtcaaccc tgctcgcaag ggcctctacg
    13801 agggcactgg ccggctcttc tttgagttct accgcctcct gcatgatgcg cggcccaagg
    13861 agggagatga tcgccccttc ttctggctct ttgagaatgt ggtggccatg ggcgttagtg
    13921 acaagaggga catctcgcga tttctcgagt ccaaccctgt gatgattgat gccaaagaag
    13981 tgtcagctgc acacagggcc cgctacttct ggggtaacct tcccggtatg aacaggccgt
    14041 tggcatccac tgtgaatgat aagctggagc tgcaggagtg tctggagcat ggcaggatag
    14101 ccaagttcag caaagtgagg accattacta cgaggtcaaa ctccataaag cagggcaaag
    14161 accagcattt tcctgtcttc atgaatgaga aagaggacat cttatggtgc actgaaatgg
    14221 aaagggtatt tggtttccca gtccactata ctgacgtctc caacatgagc cgcttggcga
    14281 ggcagagact gctgggccgg tcatggagcg tgccagtcat ccgccacctc ttcgctccgc
    14341 tgaaggagta ttttgcgtgt gtgtaagaat tcgatatcaa gcttatcgat aatcaacctc
    14401 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
    14461 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
    14521 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
    14581 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
    14641 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
    14701 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
    14761 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
    14821 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
    14881 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
    14941 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgag
    15001 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
    15061 cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt
    15121 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
    15181 gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc
    15241 acacacaagg ctacttccct gattggcaga actacacacc agggccaggg atcagatatc
    15301 cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag
    15361 ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc
    15421 cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc
    15481 gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc
    15541 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
    15601 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
    15661 agtcagtgtg gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg
    15721 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
    15781 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
    15841 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
    15901 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
    15961 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
    16021 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgct
//
