LOCUS       sequence-213362-       16287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector to express dCas9-dead 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-dead Tet1CD-P2A-BFP
FEATURES             Location/Qualifiers
     source          1..16287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        45..424
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        426..624
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     579..599
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             642..822
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    869..994
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1487..1720
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1905..1949
                     /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    2247..2364
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2429..3640
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3477..3496
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(3697..3723,3724..3726,3727..6213,6214..6216,
                     6217..7800)
                     /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             7816..7836
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7840..7860
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7864..7884
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10054..10080
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             10081..10137
                     /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             10138..10839
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    10858..11446
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10911..10931)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11329..11340)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(11449..11465)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11450..11466)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             11971..12151
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(12177..12194)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12183..12407
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      12453..12881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12540..12559)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12750..12771
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12890..12910)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12934..13144
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        12956..13152
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      13003..13138
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     13065..13084
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        13200..13247
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             13266..13640
                     /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    13770..13891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13807..13826)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13861..13880
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13940..13956)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13940..13956)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13953..13975)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13964..13980
                     /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(13988..13994,13995..14012,14013..14018))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14033..14054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14171..14188)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14342..14930)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14422..14441)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(15101..15892,15893..15961))
                     /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     15724..15743
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15962..16066)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(16141..16160)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
       61 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      121 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      181 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      241 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      301 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      361 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      421 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      481 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      541 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      601 acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc
      661 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
      721 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
      781 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
      841 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
      901 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
      961 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     1021 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     1081 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     1141 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     1201 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     1261 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     1321 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     1381 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1441 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     1501 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     1561 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     1621 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     1681 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     1741 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     1801 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     1861 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     1921 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     1981 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     2041 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     2101 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     2161 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     2221 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     2281 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     2341 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     2401 ttaattaacc cgtgtcggct ccagatctgg cctccgcgcc gggttttggc gcctcccgcg
     2461 ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg aagggcgcag cgagcgtcct
     2521 gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa
     2581 ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt
     2641 ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag
     2701 ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag
     2761 ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt
     2821 cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg
     2881 gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt
     2941 tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa
     3001 tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg
     3061 cggcaagaac ccaaggtctt gaggccttcg ctaatgcggg aaagctctta ttcgggtgag
     3121 atgggctggg gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg
     3181 gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac
     3241 ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc
     3301 agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg
     3361 ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg
     3421 ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa
     3481 gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc
     3541 accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga ctagtaaatt
     3601 gtccgctaaa ttctggccgt ttttggcttt tttgttagac gaagcttggg ctgcaggtcg
     3661 actctagagg atccagttaa cctcgagggc gcgcccatgg acaagaagta ttctatcgga
     3721 ctggccatcg ggactaatag cgtcgggtgg gccgtgatca ctgacgagta caaggtgccc
     3781 tctaagaagt tcaaggtgct cgggaacacc gaccggcatt ccatcaagaa aaatctgatc
     3841 ggagctctcc tctttgattc aggggagacc gctgaagcaa cccgcctcaa gcggactgct
     3901 agacggcggt acaccaggag gaagaaccgg atttgttacc ttcaagagat attctccaac
     3961 gaaatggcaa aggtcgacga cagcttcttc cataggctgg aagaatcatt cctcgtggaa
     4021 gaggataaga agcatgaacg gcatcccatc ttcggtaata tcgtcgacga ggtggcctat
     4081 cacgagaaat acccaaccat ctaccatctt cgcaaaaagc tggtggactc aaccgacaag
     4141 gcagacctcc ggcttatcta cctggccctg gcccacatga tcaagttcag aggccacttc
     4201 ctgatcgagg gcgacctcaa tcctgacaat agcgatgtgg ataaactgtt catccagctg
     4261 gtgcagactt acaaccagct ctttgaagag aaccccatca atgcaagcgg agtcgatgcc
     4321 aaggccattc tgtcagcccg gctgtcaaag agccgcagac ttgagaatct tatcgctcag
     4381 ctgccgggtg aaaagaaaaa tggactgttc gggaacctga ttgctctttc acttgggctg
     4441 actcccaatt tcaagtctaa tttcgacctg gcagaggatg ccaagctgca actgtccaag
     4501 gacacctatg atgacgatct cgacaacctc ctggcccaga tcggtgacca atacgccgac
     4561 cttttccttg ctgctaagaa tctttctgac gccatcctgc tgtctgacat tctccgcgtg
     4621 aacactgaaa tcaccaaggc ccctctttca gcttcaatga ttaagcggta tgatgagcac
     4681 caccaggacc tgaccctgct taaggcactc gtccggcagc agcttccgga gaagtacaag
     4741 gaaatcttct ttgaccagtc aaagaatgga tacgccggct acatcgacgg aggtgcctcc
     4801 caagaggaat tttataagtt tatcaaacct atccttgaga agatggacgg caccgaagag
     4861 ctcctcgtga aactgaatcg ggaggatctg ctgcggaagc agcgcacttt cgacaatggg
     4921 agcattcccc accagatcca tcttggggag cttcacgcca tccttcggcg ccaagaggac
     4981 ttctacccct ttcttaagga caacagggag aagattgaga aaattctcac tttccgcatc
     5041 ccctactacg tgggacccct cgccagagga aatagccggt ttgcttggat gaccagaaag
     5101 tcagaagaaa ctatcactcc ctggaacttc gaagaggtgg tggacaaggg agccagcgct
     5161 cagtcattca tcgaacggat gactaacttc gataagaacc tccccaatga gaaggtcctg
     5221 ccgaaacatt ccctgctcta cgagtacttt accgtgtaca acgagctgac caaggtgaaa
     5281 tatgtcaccg aagggatgag gaagcccgca ttcctgtcag gcgaacaaaa gaaggcaatt
     5341 gtggaccttc tgttcaagac caatagaaag gtgaccgtga agcagctgaa ggaggactat
     5401 ttcaagaaaa ttgaatgctt cgactctgtg gagattagcg gggtcgaaga tcggttcaac
     5461 gcaagcctgg gtacctacca tgatctgctt aagatcatca aggacaagga ttttctggac
     5521 aatgaggaga acgaggacat ccttgaggac attgtcctga ctctcactct gttcgaggac
     5581 cgggaaatga tcgaggagag gcttaagacc tacgcccatc tgttcgacga taaagtgatg
     5641 aagcaactta aacggagaag atataccgga tggggacgcc ttagccgcaa actcatcaac
     5701 ggaatccggg acaaacagag cggaaagacc attcttgatt tccttaaaag cgacggattc
     5761 gctaatcgca acttcatgca acttatccat gatgattccc tgacctttaa ggaggacatc
     5821 cagaaggccc aagtgtctgg acaaggtgac tcactgcacg agcatatcgc aaatctggct
     5881 ggttcacccg ctattaagaa gggtattctc cagaccgtga aagtcgtgga cgagctggtc
     5941 aaggtgatgg gtcgccataa accagagaac attgtcatcg agatggccag ggaaaaccag
     6001 actacccaga agggacagaa gaacagcagg gagcggatga aaagaattga ggaagggatt
     6061 aaggagctcg ggtcacagat ccttaaagag cacccggtgg aaaacaccca gcttcagaat
     6121 gagaagctct atctgtacta ccttcaaaat ggacgcgata tgtatgtgga ccaagagctt
     6181 gatatcaaca ggctctcaga ctacgacgtg gacgccatcg tccctcagag cttcctcaaa
     6241 gacgactcaa ttgacaataa ggtgctgact cgctcagaca agaaccgggg aaagtcagat
     6301 aacgtgccct cagaggaagt cgtgaaaaag atgaagaact attggcgcca gcttctgaac
     6361 gcaaagctga tcactcagcg gaagttcgac aatctcacta aggctgagag gggcggactg
     6421 agcgaactgg acaaagcagg attcattaaa cggcaacttg tggagactcg gcagattact
     6481 aaacatgtcg cccaaatcct tgactcacgc atgaatacca agtacgacga aaacgacaaa
     6541 cttatccgcg aggtgaaggt gattaccctg aagtccaagc tggtcagcga tttcagaaag
     6601 gactttcaat tctacaaagt gcgggagatc aataactatc atcatgctca tgacgcatat
     6661 ctgaatgccg tggtgggaac cgccctgatc aagaagtacc caaagctgga aagcgagttc
     6721 gtgtacggag actacaaggt ctacgacgtg cgcaagatga ttgccaaatc tgagcaggag
     6781 atcggaaagg ccaccgcaaa gtacttcttc tacagcaaca tcatgaattt cttcaagacc
     6841 gaaatcaccc ttgcaaacgg tgagatccgg aagaggccgc tcatcgagac taatggggag
     6901 actggcgaaa tcgtgtggga caagggcaga gatttcgcta ccgtgcgcaa agtgctttct
     6961 atgcctcaag tgaacatcgt gaagaaaacc gaggtgcaaa ccggaggctt ttctaaggaa
     7021 tcaatcctcc ccaagcgcaa ctccgacaag ctcattgcaa ggaagaagga ttgggaccct
     7081 aagaagtacg gcggattcga ttcaccaact gtggcttatt ctgtcctggt cgtggctaag
     7141 gtggaaaaag gaaagtctaa gaagctcaag agcgtgaagg aactgctggg tatcaccatt
     7201 atggagcgca gctccttcga gaagaaccca attgactttc tcgaagccaa aggttacaag
     7261 gaagtcaaga aggaccttat catcaagctc ccaaagtata gcctgttcga actggagaat
     7321 gggcggaagc ggatgctcgc ctccgctggc gaacttcaga agggtaatga gctggctctc
     7381 ccctccaagt acgtgaattt cctctacctt gcaagccatt acgagaagct gaaggggagc
     7441 cccgaggaca acgagcaaaa gcaactgttt gtggagcagc ataagcatta tctggacgag
     7501 atcattgagc agatttccga gttttctaaa cgcgtcattc tcgctgatgc caacctcgat
     7561 aaagtcctta gcgcatacaa taagcacaga gacaaaccaa ttcgggagca ggctgagaat
     7621 atcatccacc tgttcaccct caccaatctt ggtgcccctg ccgcattcaa gtacttcgac
     7681 accaccatcg accggaaacg ctatacctcc accaaagaag tgctggacgc caccctcatc
     7741 caccagagca tcaccggact ttacgaaact cggattgacc tctcacagct cggaggggat
     7801 ggtggcggag gctcgccaaa aaagaagaga aaggtagacc caaagaaaaa acgaaaagta
     7861 gatccgaaaa agaagaggaa ggtgggatcc ctgcccacct gcagctgtct tgatcgagtt
     7921 atacaaaaag acaaaggccc atattataca caccttgggg caggaccaag tgttgctgct
     7981 gtcagggaaa tcatggagaa taggtatggt caaaaaggaa acgcaataag gatagaaata
     8041 gtagtgtaca ccggtaaaga agggaaaagc tctcatgggt gtccaattgc taagtgggtt
     8101 ttaagaagaa gcagtgatga agaaaaagtt ctttgtttgg tccggcagcg tacaggccac
     8161 cactgtccaa ctgctgtgat ggtggtgctc atcatggtgt gggatggcat ccctcttcca
     8221 atggccgacc ggctatacac agagctcaca gagaatctaa agtcatacaa tgggcaccct
     8281 accgacagaa gatgcaccct caatgaaaat cgtacctgta catgtcaagg aattgatcca
     8341 gagacttgtg gagcttcatt ctcttttggc tgttcatgga gtatgtactt taatggctgt
     8401 aagtttggta gaagcccaag ccccagaaga tttagaattg atccaagctc tcccttacat
     8461 gaaaaaaacc ttgaagataa cttacagagt ttggctacac gattagctcc aatttataag
     8521 cagtatgctc cagtagctta ccaaaatcag gtggaatatg aaaatgttgc ccgagaatgt
     8581 cggcttggca gcaaggaagg tcgacccttc tctggggtca ctgcttgcct ggacttctgt
     8641 gctcatccct acagggccat tcacaacatg aataatggaa gcactgtggt ttgtacctta
     8701 actcgagaag ataaccgctc tttgggtgtt attcctcaag atgagcagct ccatgtgcta
     8761 cctctttata agctttcaga cacagatgag tttggctcca aggaaggaat ggaagccaag
     8821 atcaaatctg gggccatcga ggtcctggca ccccgccgca aaaaaagaac gtgtttcact
     8881 cagcctgttc cccgttctgg aaagaagagg gctgcgatga tgacagaggt tcttgcacat
     8941 aagataaggg cagtggaaaa gaaacctatt ccccgaatca agcggaagaa taactcaaca
     9001 acaacaaaca acagtaagcc ttcgtcactg ccaaccttag ggagtaacac tgagaccgtg
     9061 caacctgaag taaaaagtga aaccgaaccc cattttatct taaaaagttc agacaacact
     9121 aaaacttatt cgctgatgcc atccgctcct cacccagtga aagaggcatc tccaggcttc
     9181 tcctggtccc cgaagactgc ttcagccaca ccagctccac tgaagaatga cgcaacagcc
     9241 tcatgcgggt tttcagaaag aagcagcact ccccactgta cgatgccttc gggaagactc
     9301 agtggtgcca atgctgcagc tgctgatggc cctggcattt cacagcttgg cgaagtggct
     9361 cctctcccca ccctgtctgc tcctgtgatg gagcccctca ttaattctga gccttccact
     9421 ggtgtgactg agccgctaac gcctcatcag ccaaaccacc agccctcctt cctcacctct
     9481 cctcaagacc ttgcctcttc tccaatggaa gaagatgagc agcattctga agcagatgag
     9541 cctccatcag acgaacccct atctgatgac cccctgtcac ctgctgagga gaaattgccc
     9601 cacattgatg agtattggtc agacagtgag cacatctttt tggatgcaaa tattggtggg
     9661 gtggccatcg cacctgctca cggctcggtt ttgattgagt gtgcccggcg agagctgcac
     9721 gctaccactc ctgttgagca ccccaaccgt aatcatccaa cccgcctctc ccttgtcttt
     9781 taccagcaca aaaacctaaa taagccccaa catggttttg aactaaacaa gattaagttt
     9841 gaggctaaag aagctaagaa taagaaaatg aaggcctcag agcaaaaaga ccaggcagct
     9901 aatgaaggtc cagaacagtc ctctgaagta aatgaattga accaaattcc ttctcataaa
     9961 gcattaacat taacccatga caatgttgtc accgtgtccc cttatgctct cacacacgtt
    10021 gcggggccct ataaccattg ggtcgaattc gcatacccct acgacgtccc cgactacgcc
    10081 gccacgaact tctctctgtt aaagcaagca ggagatgttg aagaaaaccc cgggcctatg
    10141 agcgagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac cgtggacaac
    10201 catcacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg
    10261 agaatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctactagc
    10321 ttcctctacg gcagcaagac cttcatcaac cacacccagg gcatccccga cttcttcaag
    10381 cagtccttcc ctgagggctt cacatgggag agagtcacca catacgaaga cgggggcgtg
    10441 ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa cgtcaagatc
    10501 agaggggtga acttcacatc caacggccct gtgatgcaga agaaaacact cggctgggag
    10561 gccttcaccg agacgctgta ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc
    10621 ctgaagctcg tgggcgggag ccatctgatc gcaaacatca agaccacata tagatccaag
    10681 aaacccgcta agaacctcaa gatgcctggc gtctactatg tggactacag actggaaaga
    10741 atcaaggagg ccaacaacga gacctacgtc gagcagcacg aggtggcagt ggccagatac
    10801 tgcgacctcc ctagcaaact ggggcacaag cttaattgag atatcaagct tatcgataat
    10861 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
    10921 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
    10981 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
    11041 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
    11101 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
    11161 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
    11221 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
    11281 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
    11341 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
    11401 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga
    11461 cctcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct
    11521 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag
    11581 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
    11641 aaggggggac tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg
    11701 atctaccaca cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc
    11761 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta
    11821 gaagaagcca atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg
    11881 gatgacccgg agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac
    11941 atggcccgag agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc
    12001 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
    12061 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
    12121 cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc
    12181 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
    12241 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
    12301 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
    12361 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
    12421 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
    12481 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
    12541 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
    12601 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
    12661 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
    12721 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
    12781 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
    12841 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
    12901 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
    12961 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
    13021 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
    13081 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
    13141 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
    13201 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
    13261 aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag
    13321 cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg
    13381 ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc
    13441 cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt
    13501 cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg
    13561 agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg
    13621 tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg
    13681 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
    13741 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
    13801 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
    13861 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
    13921 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
    13981 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
    14041 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
    14101 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
    14161 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
    14221 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
    14281 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
    14341 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
    14401 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
    14461 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
    14521 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
    14581 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
    14641 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
    14701 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
    14761 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
    14821 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
    14881 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
    14941 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
    15001 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
    15061 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
    15121 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
    15181 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
    15241 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
    15301 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
    15361 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
    15421 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
    15481 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
    15541 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
    15601 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
    15661 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
    15721 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
    15781 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
    15841 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
    15901 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
    15961 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
    16021 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
    16081 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
    16141 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
    16201 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
    16261 ttgcatgaag aatctgctta gggttag
//
