LOCUS       sequence-306393-       16869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct to express dCas9 fused with Dnmt3a-P2A-tagBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16869)
  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 16869)
  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-P2A-tagBFP
FEATURES             Location/Qualifiers
     source          1..16869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1212
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1049..1068
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(1269..1295,1296..1298,1299..3785,3786..3788,
                     3789..5372)
                     /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             5388..5408
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5412..5432
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5436..5456
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5463..8198
                     /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"
     CDS             8208..8234
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             8235..8291
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             8292..8993
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    9012..9600
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9065..9085)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9483..9494)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9603..9619)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9604..9620)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10125..10305
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10331..10348)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10337..10561
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10607..11035
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10694..10713)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10904..10925
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11044..11064)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11088..11298
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        11110..11306
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      11157..11292
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11219..11238
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11354..11401
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11420..11794
                     /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    11924..12045
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11961..11980)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12015..12034
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12094..12110)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12094..12110)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12107..12129)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12118..12134
                     /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(12142..12148,12149..12166,12167..12172))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12187..12208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12325..12342)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12496..13084)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12576..12595)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13255..14046,14047..14115))
                     /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     13878..13897
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14116..14220)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14295..14314)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        14486..14865
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        14867..15065
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     15020..15040
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             15083..15263
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    15310..15435
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    15928..16161
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             16346..16390
                     /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    16688..16805
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg
       61 ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg ctcaggacag
      121 cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag
      181 gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg
      241 aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat
      301 gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt
      361 cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct
      421 ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc
      481 caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg
      541 cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga
      601 ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgaggcctt
      661 cgctaatgcg ggaaagctct tattcgggtg agatgggctg gggcaccatc tggggaccct
      721 gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt
      781 tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc
      841 gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg
      901 cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat
      961 cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg
     1021 gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg
     1081 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca
     1141 atatgtaatt ttcagtgtta gactagtaaa ttgtccgcta aattctggcc gtttttggct
     1201 tttttgttag acgaagcttg ggctgcaggt cgactctaga ggatccagtt aacctcgagg
     1261 gcgcgcccat ggacaagaag tattctatcg gactggccat cgggactaat agcgtcgggt
     1321 gggccgtgat cactgacgag tacaaggtgc cctctaagaa gttcaaggtg ctcgggaaca
     1381 ccgaccggca ttccatcaag aaaaatctga tcggagctct cctctttgat tcaggggaga
     1441 ccgctgaagc aacccgcctc aagcggactg ctagacggcg gtacaccagg aggaagaacc
     1501 ggatttgtta ccttcaagag atattctcca acgaaatggc aaaggtcgac gacagcttct
     1561 tccataggct ggaagaatca ttcctcgtgg aagaggataa gaagcatgaa cggcatccca
     1621 tcttcggtaa tatcgtcgac gaggtggcct atcacgagaa atacccaacc atctaccatc
     1681 ttcgcaaaaa gctggtggac tcaaccgaca aggcagacct ccggcttatc tacctggccc
     1741 tggcccacat gatcaagttc agaggccact tcctgatcga gggcgacctc aatcctgaca
     1801 atagcgatgt ggataaactg ttcatccagc tggtgcagac ttacaaccag ctctttgaag
     1861 agaaccccat caatgcaagc ggagtcgatg ccaaggccat tctgtcagcc cggctgtcaa
     1921 agagccgcag acttgagaat cttatcgctc agctgccggg tgaaaagaaa aatggactgt
     1981 tcgggaacct gattgctctt tcacttgggc tgactcccaa tttcaagtct aatttcgacc
     2041 tggcagagga tgccaagctg caactgtcca aggacaccta tgatgacgat ctcgacaacc
     2101 tcctggccca gatcggtgac caatacgccg accttttcct tgctgctaag aatctttctg
     2161 acgccatcct gctgtctgac attctccgcg tgaacactga aatcaccaag gcccctcttt
     2221 cagcttcaat gattaagcgg tatgatgagc accaccagga cctgaccctg cttaaggcac
     2281 tcgtccggca gcagcttccg gagaagtaca aggaaatctt ctttgaccag tcaaagaatg
     2341 gatacgccgg ctacatcgac ggaggtgcct cccaagagga attttataag tttatcaaac
     2401 ctatccttga gaagatggac ggcaccgaag agctcctcgt gaaactgaat cgggaggatc
     2461 tgctgcggaa gcagcgcact ttcgacaatg ggagcattcc ccaccagatc catcttgggg
     2521 agcttcacgc catccttcgg cgccaagagg acttctaccc ctttcttaag gacaacaggg
     2581 agaagattga gaaaattctc actttccgca tcccctacta cgtgggaccc ctcgccagag
     2641 gaaatagccg gtttgcttgg atgaccagaa agtcagaaga aactatcact ccctggaact
     2701 tcgaagaggt ggtggacaag ggagccagcg ctcagtcatt catcgaacgg atgactaact
     2761 tcgataagaa cctccccaat gagaaggtcc tgccgaaaca ttccctgctc tacgagtact
     2821 ttaccgtgta caacgagctg accaaggtga aatatgtcac cgaagggatg aggaagcccg
     2881 cattcctgtc aggcgaacaa aagaaggcaa ttgtggacct tctgttcaag accaatagaa
     2941 aggtgaccgt gaagcagctg aaggaggact atttcaagaa aattgaatgc ttcgactctg
     3001 tggagattag cggggtcgaa gatcggttca acgcaagcct gggtacctac catgatctgc
     3061 ttaagatcat caaggacaag gattttctgg acaatgagga gaacgaggac atccttgagg
     3121 acattgtcct gactctcact ctgttcgagg accgggaaat gatcgaggag aggcttaaga
     3181 cctacgccca tctgttcgac gataaagtga tgaagcaact taaacggaga agatataccg
     3241 gatggggacg ccttagccgc aaactcatca acggaatccg ggacaaacag agcggaaaga
     3301 ccattcttga tttccttaaa agcgacggat tcgctaatcg caacttcatg caacttatcc
     3361 atgatgattc cctgaccttt aaggaggaca tccagaaggc ccaagtgtct ggacaaggtg
     3421 actcactgca cgagcatatc gcaaatctgg ctggttcacc cgctattaag aagggtattc
     3481 tccagaccgt gaaagtcgtg gacgagctgg tcaaggtgat gggtcgccat aaaccagaga
     3541 acattgtcat cgagatggcc agggaaaacc agactaccca gaagggacag aagaacagca
     3601 gggagcggat gaaaagaatt gaggaaggga ttaaggagct cgggtcacag atccttaaag
     3661 agcacccggt ggaaaacacc cagcttcaga atgagaagct ctatctgtac taccttcaaa
     3721 atggacgcga tatgtatgtg gaccaagagc ttgatatcaa caggctctca gactacgacg
     3781 tggacgccat cgtccctcag agcttcctca aagacgactc aattgacaat aaggtgctga
     3841 ctcgctcaga caagaaccgg ggaaagtcag ataacgtgcc ctcagaggaa gtcgtgaaaa
     3901 agatgaagaa ctattggcgc cagcttctga acgcaaagct gatcactcag cggaagttcg
     3961 acaatctcac taaggctgag aggggcggac tgagcgaact ggacaaagca ggattcatta
     4021 aacggcaact tgtggagact cggcagatta ctaaacatgt cgcccaaatc cttgactcac
     4081 gcatgaatac caagtacgac gaaaacgaca aacttatccg cgaggtgaag gtgattaccc
     4141 tgaagtccaa gctggtcagc gatttcagaa aggactttca attctacaaa gtgcgggaga
     4201 tcaataacta tcatcatgct catgacgcat atctgaatgc cgtggtggga accgccctga
     4261 tcaagaagta cccaaagctg gaaagcgagt tcgtgtacgg agactacaag gtctacgacg
     4321 tgcgcaagat gattgccaaa tctgagcagg agatcggaaa ggccaccgca aagtacttct
     4381 tctacagcaa catcatgaat ttcttcaaga ccgaaatcac ccttgcaaac ggtgagatcc
     4441 ggaagaggcc gctcatcgag actaatgggg agactggcga aatcgtgtgg gacaagggca
     4501 gagatttcgc taccgtgcgc aaagtgcttt ctatgcctca agtgaacatc gtgaagaaaa
     4561 ccgaggtgca aaccggaggc ttttctaagg aatcaatcct ccccaagcgc aactccgaca
     4621 agctcattgc aaggaagaag gattgggacc ctaagaagta cggcggattc gattcaccaa
     4681 ctgtggctta ttctgtcctg gtcgtggcta aggtggaaaa aggaaagtct aagaagctca
     4741 agagcgtgaa ggaactgctg ggtatcacca ttatggagcg cagctccttc gagaagaacc
     4801 caattgactt tctcgaagcc aaaggttaca aggaagtcaa gaaggacctt atcatcaagc
     4861 tcccaaagta tagcctgttc gaactggaga atgggcggaa gcggatgctc gcctccgctg
     4921 gcgaacttca gaagggtaat gagctggctc tcccctccaa gtacgtgaat ttcctctacc
     4981 ttgcaagcca ttacgagaag ctgaagggga gccccgagga caacgagcaa aagcaactgt
     5041 ttgtggagca gcataagcat tatctggacg agatcattga gcagatttcc gagttttcta
     5101 aacgcgtcat tctcgctgat gccaacctcg ataaagtcct tagcgcatac aataagcaca
     5161 gagacaaacc aattcgggag caggctgaga atatcatcca cctgttcacc ctcaccaatc
     5221 ttggtgcccc tgccgcattc aagtacttcg acaccaccat cgaccggaaa cgctatacct
     5281 ccaccaaaga agtgctggac gccaccctca tccaccagag catcaccgga ctttacgaaa
     5341 ctcggattga cctctcacag ctcggagggg atggtggcgg aggctcgcca aaaaagaaga
     5401 gaaaggtaga cccaaagaaa aaacgaaaag tagatccgaa aaagaagagg aaggtgggat
     5461 ccatgcccgc catgccctcc agcggccccg gggacaccag cagctctgct gcggagcggg
     5521 aggaggaccg aaaggacgga gaggagcagg aggagccgcg tggcaaggag gagcgccaag
     5581 agcccagcac cacggcacgg aaggtggggc ggcctgggag gaagcgcaag caccccccgg
     5641 tggaaagcgg tgacacgcca aaggaccctg cggtgatctc caagtcccca tccatggccc
     5701 aggactcagg cgcctcagag ctattaccca atggggactt ggagaagcgg agtgagcccc
     5761 agccagagga ggggagccct gctggggggc agaagggcgg ggccccagca gagggagagg
     5821 gtgcagctga gaccctgcct gaagcctcaa gagcagtgga aaatggctgc tgcaccccca
     5881 aggagggccg aggagcccct gcagaagcgg gcaaagaaca gaaggagacc aacatcgaat
     5941 ccatgaaaat ggagggctcc cggggccggc tgcggggtgg cttgggctgg gagtccagcc
     6001 tccgtcagcg gcccatgccg aggctcacct tccaggcggg ggacccctac tacatcagca
     6061 agcgcaagcg ggacgagtgg ctggcacgct ggaaaaggga ggctgagaag aaagccaagg
     6121 tcattgcagg aatgaatgct gtggaagaaa accaggggcc cggggagtct cagaaggtgg
     6181 aggaggccag ccctcctgct gtgcagcagc ccactgaccc cgcatccccc actgtggcta
     6241 ccacgcctga gcccgtgggg tccgatgctg gggacaagaa tgccaccaaa gcaggcgatg
     6301 acgagccaga gtacgaggac ggccggggct ttggcattgg ggagctggtg tgggggaaac
     6361 tgcggggctt ctcctggtgg ccaggccgca ttgtgtcttg gtggatgacg ggccggagcc
     6421 gagcagctga aggcacccgc tgggtcatgt ggttcggaga cggcaaattc tcagtggtgt
     6481 gtgttgagaa gctgatgccg ctgagctcgt tttgcagtgc gttccaccag gccacgtaca
     6541 acaagcagcc catgtaccgc aaagccatct acgaggtcct gcaggtggcc agcagccgcg
     6601 cggggaagct gttcccggtg tgccacgaca gcgatgagag tgacactgcc aaggccgtgg
     6661 aggtgcagaa caagcccatg attgaatggg ccctgggggg cttccagcct tctggcccta
     6721 agggcctgga gccaccagaa gaagagaaga atccctacaa agaagtgtac acggacatgt
     6781 gggtggaacc tgaggcagct gcctacgcac cacctccacc agccaaaaag ccccggaaga
     6841 gcacagcgga gaagcccaag gtcaaggaga ttattgatga gcgcacaaga gagcggctgg
     6901 tgtacgaggt gcggcagaag tgccggaaca ttgaggacat ctgcatctcc tgtgggagcc
     6961 tcaatgttac cctggaacac cccctcttcg ttggaggaat gtgccaaaac tgcaagaact
     7021 gctttctgga gtgtgcgtac cagtacgacg acgacggcta ccagtcctac tgcaccatct
     7081 gctgtggggg ccgtgaggtg ctcatgtgcg gaaacaacaa ctgctgcagg tgcttttgcg
     7141 tggagtgtgt ggacctcttg gtggggccgg gggctgccca ggcagccatt aaggaagacc
     7201 cctggaactg ctacatgtgc gggcacaagg gtacctacgg gctgctgcgg cggcgagagg
     7261 actggccctc ccggctccag atgttcttcg ctaataacca cgaccaggaa tttgaccctc
     7321 caaaggttta cccacctgtc ccagctgaga agaggaagcc catccgggtg ctgtctctct
     7381 ttgatggaat cgctacaggg ctcctggtgc tgaaggactt gggcattcag gtggaccgct
     7441 acattgcctc ggaggtgtgt gaggactcca tcacggtggg catggtgcgg caccagggga
     7501 agatcatgta cgtcggggac gtccgcagcg tcacacagaa gcatatccag gagtggggcc
     7561 cattcgatct ggtgattggg ggcagtccct gcaatgacct ctccatcgtc aaccctgctc
     7621 gcaagggcct ctacgagggc actggccggc tcttctttga gttctaccgc ctcctgcatg
     7681 atgcgcggcc caaggaggga gatgatcgcc ccttcttctg gctctttgag aatgtggtgg
     7741 ccatgggcgt tagtgacaag agggacatct cgcgatttct cgagtccaac cctgtgatga
     7801 ttgatgccaa agaagtgtca gctgcacaca gggcccgcta cttctggggt aaccttcccg
     7861 gtatgaacag gccgttggca tccactgtga atgataagct ggagctgcag gagtgtctgg
     7921 agcatggcag gatagccaag ttcagcaaag tgaggaccat tactacgagg tcaaactcca
     7981 taaagcaggg caaagaccag cattttcctg tcttcatgaa tgagaaagag gacatcttat
     8041 ggtgcactga aatggaaagg gtatttggtt tcccagtcca ctatactgac gtctccaaca
     8101 tgagccgctt ggcgaggcag agactgctgg gccggtcatg gagcgtgcca gtcatccgcc
     8161 acctcttcgc tccgctgaag gagtattttg cgtgtgtgga attcgcatac ccctacgacg
     8221 tccccgacta cgccgccacg aacttctctc tgttaaagca agcaggagat gttgaagaaa
     8281 accccgggcc tatgagcgag ctgattaagg agaacatgca catgaagctg tacatggagg
     8341 gcaccgtgga caaccatcac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg
     8401 gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca
     8461 tcctggctac tagcttcctc tacggcagca agaccttcat caaccacacc cagggcatcc
     8521 ccgacttctt caagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg
     8581 aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct
     8641 acaacgtcaa gatcagaggg gtgaacttca catccaacgg ccctgtgatg cagaagaaaa
     8701 cactcggctg ggaggccttc accgagacgc tgtaccccgc tgacggcggc ctggaaggca
     8761 gaaacgacat ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac atcaagacca
     8821 catatagatc caagaaaccc gctaagaacc tcaagatgcc tggcgtctac tatgtggact
     8881 acagactgga aagaatcaag gaggccaaca acgagaccta cgtcgagcag cacgaggtgg
     8941 cagtggccag atactgcgac ctccctagca aactggggca caagcttaat tgagatatca
     9001 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     9061 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     9121 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     9181 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     9241 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     9301 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     9361 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     9421 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     9481 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     9541 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     9601 atcgataccg tcgacctcga gacctagaaa aacatggagc aatcacaagt agcaatacag
     9661 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc
     9721 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     9781 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata
     9841 tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag aactacacac
     9901 cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg
     9961 agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga
    10021 gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc
    10081 tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag
    10141 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
    10201 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
    10261 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc
    10321 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
    10381 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
    10441 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
    10501 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
    10561 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
    10621 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
    10681 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
    10741 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
    10801 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
    10861 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
    10921 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
    10981 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
    11041 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
    11101 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
    11161 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
    11221 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
    11281 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
    11341 ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
    11401 caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg
    11461 cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt
    11521 ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca
    11581 ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct
    11641 gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat
    11701 gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa
    11761 ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac
    11821 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
    11881 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
    11941 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
    12001 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
    12061 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
    12121 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
    12181 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
    12241 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
    12301 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
    12361 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
    12421 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
    12481 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
    12541 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
    12601 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
    12661 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
    12721 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
    12781 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
    12841 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
    12901 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
    12961 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
    13021 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
    13081 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
    13141 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
    13201 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
    13261 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
    13321 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
    13381 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
    13441 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
    13501 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
    13561 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
    13621 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
    13681 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
    13741 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
    13801 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
    13861 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
    13921 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
    13981 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
    14041 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
    14101 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
    14161 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
    14221 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
    14281 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
    14341 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
    14401 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
    14461 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
    14521 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
    14581 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
    14641 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
    14701 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
    14761 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
    14821 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
    14881 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
    14941 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
    15001 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
    15061 agcagcgcgt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
    15121 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
    15181 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
    15241 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
    15301 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
    15361 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
    15421 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
    15481 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
    15541 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
    15601 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
    15661 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
    15721 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct
    15781 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
    15841 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
    15901 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
    15961 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
    16021 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
    16081 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
    16141 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
    16201 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
    16261 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
    16321 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
    16381 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
    16441 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
    16501 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
    16561 acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat
    16621 tcgattagtg aacggatcgg cactgcgtgc gccaattctg cagacaaatg gcagtattca
    16681 tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
    16741 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
    16801 attttcgggt ttattacagg gacagcagag atccagtttg gttaattaac ccgtgtcggc
    16861 tccagatct
//
