LOCUS       sequence-207911-       16287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector to express dCas9-Tet1CD-P2A-tagBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16287)
  AUTHORS   Liu XS, Wu H, Krzisch M, Wu X, Graef J, Muffat J, Hnisz D, Li CH, 
            Yuan B, Xu C, Li Y, Vershkov D, Cacace A, Young RA, Jaenisch R
  TITLE     Rescue of Fragile X Syndrome Neurons by DNA Methylation Editing of 
            the FMR1 Gene.
  JOURNAL   Cell. 2018 Feb 8. pii: S0092-8674(18)30049-7. doi: 
            10.1016/j.cell.2018.01.012.
  PUBMED    29456084
REFERENCE   2  (bases 1 to 16287)
  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: "Cell. 2018 
            Feb 8. pii: S0092-8674(18)30049-7. doi: 10.1016/j.cell.2018.01.012."
COMMENT     Sequence Label: Fuw-dCas9-Tet1CD-P2A-BFP
FEATURES             Location/Qualifiers
     source          1..16287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(52..71)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        243..622
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        624..822
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     777..797
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             840..1020
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1067..1192
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1685..1918
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2103..2147
                     /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    2445..2562
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2627..3838
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3675..3694
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(3895..3921,3922..3924,3925..6411,6412..6414,
                     6415..7998)
                     /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             8014..8034
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8038..8058
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8062..8082
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10252..10278
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             10279..10335
                     /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             10336..11037
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    11056..11644
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11109..11129)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11527..11538)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(11647..11663)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11648..11664)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             12169..12349
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(12375..12392)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12381..12605
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      12651..13079
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12738..12757)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12948..12969
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(13088..13108)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        13132..13342
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        13154..13350
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      13201..13336
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     13263..13282
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        13398..13445
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             13464..13838
                     /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    13968..14089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(14005..14024)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     14059..14078
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(14138..14154)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14138..14154)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14151..14173)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14162..14178
                     /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(14186..14192,14193..14210,14211..14216))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14231..14252
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14369..14386)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14540..15128)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14620..14639)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(15299..16090,16091..16159))
                     /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     15922..15941
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(16160..16264)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
       61 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
      121 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
      181 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
      241 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
      301 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      361 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      421 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      481 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      541 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      601 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      661 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      721 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      781 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt gcctgtactg
      841 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
      901 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
      961 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     1021 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
     1081 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
     1141 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
     1201 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
     1261 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     1321 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     1381 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     1441 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     1501 aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag
     1561 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     1621 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     1681 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     1741 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     1801 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     1861 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     1921 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     1981 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     2041 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     2101 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     2161 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     2221 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     2281 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     2341 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
     2401 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     2461 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     2521 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     2581 agcagagatc cagtttggtt aattaacccg tgtcggctcc agatctggcc tccgcgccgg
     2641 gttttggcgc ctcccgcggg cgcccccctc ctcacggcga gcgctgccac gtcagacgaa
     2701 gggcgcagcg agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca
     2761 taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt
     2821 gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt
     2881 ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac
     2941 gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt
     3001 gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg
     3061 tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc
     3121 tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg
     3181 ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa
     3241 ggtggggggc atggtgggcg gcaagaaccc aaggtcttga ggccttcgct aatgcgggaa
     3301 agctcttatt cgggtgagat gggctggggc accatctggg gaccctgacg tgaagtttgt
     3361 cactgactgg agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt
     3421 tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt
     3481 tctgttggct tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct
     3541 ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga
     3601 cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta
     3661 tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt
     3721 ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca
     3781 gtgttagact agtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacga
     3841 agcttgggct gcaggtcgac tctagaggat ccagttaacc tcgagggcgc gcccatggac
     3901 aagaagtatt ctatcggact ggccatcggg actaatagcg tcgggtgggc cgtgatcact
     3961 gacgagtaca aggtgccctc taagaagttc aaggtgctcg ggaacaccga ccggcattcc
     4021 atcaagaaaa atctgatcgg agctctcctc tttgattcag gggagaccgc tgaagcaacc
     4081 cgcctcaagc ggactgctag acggcggtac accaggagga agaaccggat ttgttacctt
     4141 caagagatat tctccaacga aatggcaaag gtcgacgaca gcttcttcca taggctggaa
     4201 gaatcattcc tcgtggaaga ggataagaag catgaacggc atcccatctt cggtaatatc
     4261 gtcgacgagg tggcctatca cgagaaatac ccaaccatct accatcttcg caaaaagctg
     4321 gtggactcaa ccgacaaggc agacctccgg cttatctacc tggccctggc ccacatgatc
     4381 aagttcagag gccacttcct gatcgagggc gacctcaatc ctgacaatag cgatgtggat
     4441 aaactgttca tccagctggt gcagacttac aaccagctct ttgaagagaa ccccatcaat
     4501 gcaagcggag tcgatgccaa ggccattctg tcagcccggc tgtcaaagag ccgcagactt
     4561 gagaatctta tcgctcagct gccgggtgaa aagaaaaatg gactgttcgg gaacctgatt
     4621 gctctttcac ttgggctgac tcccaatttc aagtctaatt tcgacctggc agaggatgcc
     4681 aagctgcaac tgtccaagga cacctatgat gacgatctcg acaacctcct ggcccagatc
     4741 ggtgaccaat acgccgacct tttccttgct gctaagaatc tttctgacgc catcctgctg
     4801 tctgacattc tccgcgtgaa cactgaaatc accaaggccc ctctttcagc ttcaatgatt
     4861 aagcggtatg atgagcacca ccaggacctg accctgctta aggcactcgt ccggcagcag
     4921 cttccggaga agtacaagga aatcttcttt gaccagtcaa agaatggata cgccggctac
     4981 atcgacggag gtgcctccca agaggaattt tataagttta tcaaacctat ccttgagaag
     5041 atggacggca ccgaagagct cctcgtgaaa ctgaatcggg aggatctgct gcggaagcag
     5101 cgcactttcg acaatgggag cattccccac cagatccatc ttggggagct tcacgccatc
     5161 cttcggcgcc aagaggactt ctaccccttt cttaaggaca acagggagaa gattgagaaa
     5221 attctcactt tccgcatccc ctactacgtg ggacccctcg ccagaggaaa tagccggttt
     5281 gcttggatga ccagaaagtc agaagaaact atcactccct ggaacttcga agaggtggtg
     5341 gacaagggag ccagcgctca gtcattcatc gaacggatga ctaacttcga taagaacctc
     5401 cccaatgaga aggtcctgcc gaaacattcc ctgctctacg agtactttac cgtgtacaac
     5461 gagctgacca aggtgaaata tgtcaccgaa gggatgagga agcccgcatt cctgtcaggc
     5521 gaacaaaaga aggcaattgt ggaccttctg ttcaagacca atagaaaggt gaccgtgaag
     5581 cagctgaagg aggactattt caagaaaatt gaatgcttcg actctgtgga gattagcggg
     5641 gtcgaagatc ggttcaacgc aagcctgggt acctaccatg atctgcttaa gatcatcaag
     5701 gacaaggatt ttctggacaa tgaggagaac gaggacatcc ttgaggacat tgtcctgact
     5761 ctcactctgt tcgaggaccg ggaaatgatc gaggagaggc ttaagaccta cgcccatctg
     5821 ttcgacgata aagtgatgaa gcaacttaaa cggagaagat ataccggatg gggacgcctt
     5881 agccgcaaac tcatcaacgg aatccgggac aaacagagcg gaaagaccat tcttgatttc
     5941 cttaaaagcg acggattcgc taatcgcaac ttcatgcaac ttatccatga tgattccctg
     6001 acctttaagg aggacatcca gaaggcccaa gtgtctggac aaggtgactc actgcacgag
     6061 catatcgcaa atctggctgg ttcacccgct attaagaagg gtattctcca gaccgtgaaa
     6121 gtcgtggacg agctggtcaa ggtgatgggt cgccataaac cagagaacat tgtcatcgag
     6181 atggccaggg aaaaccagac tacccagaag ggacagaaga acagcaggga gcggatgaaa
     6241 agaattgagg aagggattaa ggagctcggg tcacagatcc ttaaagagca cccggtggaa
     6301 aacacccagc ttcagaatga gaagctctat ctgtactacc ttcaaaatgg acgcgatatg
     6361 tatgtggacc aagagcttga tatcaacagg ctctcagact acgacgtgga cgccatcgtc
     6421 cctcagagct tcctcaaaga cgactcaatt gacaataagg tgctgactcg ctcagacaag
     6481 aaccggggaa agtcagataa cgtgccctca gaggaagtcg tgaaaaagat gaagaactat
     6541 tggcgccagc ttctgaacgc aaagctgatc actcagcgga agttcgacaa tctcactaag
     6601 gctgagaggg gcggactgag cgaactggac aaagcaggat tcattaaacg gcaacttgtg
     6661 gagactcggc agattactaa acatgtcgcc caaatccttg actcacgcat gaataccaag
     6721 tacgacgaaa acgacaaact tatccgcgag gtgaaggtga ttaccctgaa gtccaagctg
     6781 gtcagcgatt tcagaaagga ctttcaattc tacaaagtgc gggagatcaa taactatcat
     6841 catgctcatg acgcatatct gaatgccgtg gtgggaaccg ccctgatcaa gaagtaccca
     6901 aagctggaaa gcgagttcgt gtacggagac tacaaggtct acgacgtgcg caagatgatt
     6961 gccaaatctg agcaggagat cggaaaggcc accgcaaagt acttcttcta cagcaacatc
     7021 atgaatttct tcaagaccga aatcaccctt gcaaacggtg agatccggaa gaggccgctc
     7081 atcgagacta atggggagac tggcgaaatc gtgtgggaca agggcagaga tttcgctacc
     7141 gtgcgcaaag tgctttctat gcctcaagtg aacatcgtga agaaaaccga ggtgcaaacc
     7201 ggaggctttt ctaaggaatc aatcctcccc aagcgcaact ccgacaagct cattgcaagg
     7261 aagaaggatt gggaccctaa gaagtacggc ggattcgatt caccaactgt ggcttattct
     7321 gtcctggtcg tggctaaggt ggaaaaagga aagtctaaga agctcaagag cgtgaaggaa
     7381 ctgctgggta tcaccattat ggagcgcagc tccttcgaga agaacccaat tgactttctc
     7441 gaagccaaag gttacaagga agtcaagaag gaccttatca tcaagctccc aaagtatagc
     7501 ctgttcgaac tggagaatgg gcggaagcgg atgctcgcct ccgctggcga acttcagaag
     7561 ggtaatgagc tggctctccc ctccaagtac gtgaatttcc tctaccttgc aagccattac
     7621 gagaagctga aggggagccc cgaggacaac gagcaaaagc aactgtttgt ggagcagcat
     7681 aagcattatc tggacgagat cattgagcag atttccgagt tttctaaacg cgtcattctc
     7741 gctgatgcca acctcgataa agtccttagc gcatacaata agcacagaga caaaccaatt
     7801 cgggagcagg ctgagaatat catccacctg ttcaccctca ccaatcttgg tgcccctgcc
     7861 gcattcaagt acttcgacac caccatcgac cggaaacgct atacctccac caaagaagtg
     7921 ctggacgcca ccctcatcca ccagagcatc accggacttt acgaaactcg gattgacctc
     7981 tcacagctcg gaggggatgg tggcggaggc tcgccaaaaa agaagagaaa ggtagaccca
     8041 aagaaaaaac gaaaagtaga tccgaaaaag aagaggaagg tgggatccct gcccacctgc
     8101 agctgtcttg atcgagttat acaaaaagac aaaggcccat attatacaca ccttggggca
     8161 ggaccaagtg ttgctgctgt cagggaaatc atggagaata ggtatggtca aaaaggaaac
     8221 gcaataagga tagaaatagt agtgtacacc ggtaaagaag ggaaaagctc tcatgggtgt
     8281 ccaattgcta agtgggtttt aagaagaagc agtgatgaag aaaaagttct ttgtttggtc
     8341 cggcagcgta caggccacca ctgtccaact gctgtgatgg tggtgctcat catggtgtgg
     8401 gatggcatcc ctcttccaat ggccgaccgg ctatacacag agctcacaga gaatctaaag
     8461 tcatacaatg ggcaccctac cgacagaaga tgcaccctca atgaaaatcg tacctgtaca
     8521 tgtcaaggaa ttgatccaga gacttgtgga gcttcattct cttttggctg ttcatggagt
     8581 atgtacttta atggctgtaa gtttggtaga agcccaagcc ccagaagatt tagaattgat
     8641 ccaagctctc ccttacatga aaaaaacctt gaagataact tacagagttt ggctacacga
     8701 ttagctccaa tttataagca gtatgctcca gtagcttacc aaaatcaggt ggaatatgaa
     8761 aatgttgccc gagaatgtcg gcttggcagc aaggaaggtc gacccttctc tggggtcact
     8821 gcttgcctgg acttctgtgc tcatccccac agggacattc acaacatgaa taatggaagc
     8881 actgtggttt gtaccttaac tcgagaagat aaccgctctt tgggtgttat tcctcaagat
     8941 gagcagctcc atgtgctacc tctttataag ctttcagaca cagatgagtt tggctccaag
     9001 gaaggaatgg aagccaagat caaatctggg gccatcgagg tcctggcacc ccgccgcaaa
     9061 aaaagaacgt gtttcactca gcctgttccc cgttctggaa agaagagggc tgcgatgatg
     9121 acagaggttc ttgcacataa gataagggca gtggaaaaga aacctattcc ccgaatcaag
     9181 cggaagaata actcaacaac aacaaacaac agtaagcctt cgtcactgcc aaccttaggg
     9241 agtaacactg agaccgtgca acctgaagta aaaagtgaaa ccgaacccca ttttatctta
     9301 aaaagttcag acaacactaa aacttattcg ctgatgccat ccgctcctca cccagtgaaa
     9361 gaggcatctc caggcttctc ctggtccccg aagactgctt cagccacacc agctccactg
     9421 aagaatgacg caacagcctc atgcgggttt tcagaaagaa gcagcactcc ccactgtacg
     9481 atgccttcgg gaagactcag tggtgccaat gctgcagctg ctgatggccc tggcatttca
     9541 cagcttggcg aagtggctcc tctccccacc ctgtctgctc ctgtgatgga gcccctcatt
     9601 aattctgagc cttccactgg tgtgactgag ccgctaacgc ctcatcagcc aaaccaccag
     9661 ccctccttcc tcacctctcc tcaagacctt gcctcttctc caatggaaga agatgagcag
     9721 cattctgaag cagatgagcc tccatcagac gaacccctat ctgatgaccc cctgtcacct
     9781 gctgaggaga aattgcccca cattgatgag tattggtcag acagtgagca catctttttg
     9841 gatgcaaata ttggtggggt ggccatcgca cctgctcacg gctcggtttt gattgagtgt
     9901 gcccggcgag agctgcacgc taccactcct gttgagcacc ccaaccgtaa tcatccaacc
     9961 cgcctctccc ttgtctttta ccagcacaaa aacctaaata agccccaaca tggttttgaa
    10021 ctaaacaaga ttaagtttga ggctaaagaa gctaagaata agaaaatgaa ggcctcagag
    10081 caaaaagacc aggcagctaa tgaaggtcca gaacagtcct ctgaagtaaa tgaattgaac
    10141 caaattcctt ctcataaagc attaacatta acccatgaca atgttgtcac cgtgtcccct
    10201 tatgctctca cacacgttgc ggggccctat aaccattggg tcgaattcgc atacccctac
    10261 gacgtccccg actacgccgc cacgaacttc tctctgttaa agcaagcagg agatgttgaa
    10321 gaaaaccccg ggcctatgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg
    10381 gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac
    10441 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
    10501 gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc
    10561 atccccgact tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca
    10621 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
    10681 atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag
    10741 aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa
    10801 ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacatcaag
    10861 accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg
    10921 gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag
    10981 gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct taattgagat
    11041 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
    11101 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
    11161 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
    11221 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
    11281 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
    11341 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
    11401 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
    11461 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
    11521 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
    11581 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
    11641 ccgcatcgat accgtcgacc tcgagaccta gaaaaacatg gagcaatcac aagtagcaat
    11701 acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt
    11761 tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
    11821 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
    11881 gatatccttg atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac
    11941 acaccagggc cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca
    12001 gttgagcaag agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct
    12061 gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc
    12121 cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg gtctctctgg
    12181 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
    12241 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
    12301 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta
    12361 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
    12421 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
    12481 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
    12541 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
    12601 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
    12661 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
    12721 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
    12781 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
    12841 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
    12901 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
    12961 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
    13021 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
    13081 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
    13141 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
    13201 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
    13261 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
    13321 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
    13381 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
    13441 acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg
    13501 cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact
    13561 tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg
    13621 tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg
    13681 agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg
    13741 ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg
    13801 gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt
    13861 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
    13921 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
    13981 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
    14041 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
    14101 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
    14161 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
    14221 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
    14281 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
    14341 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
    14401 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
    14461 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
    14521 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
    14581 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
    14641 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
    14701 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
    14761 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
    14821 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
    14881 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
    14941 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
    15001 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
    15061 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
    15121 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
    15181 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
    15241 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
    15301 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
    15361 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
    15421 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
    15481 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
    15541 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
    15601 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
    15661 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
    15721 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
    15781 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
    15841 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
    15901 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
    15961 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
    16021 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
    16081 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
    16141 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
    16201 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
    16261 cgcgcacatt tccccgaaaa gtgccac
//
