LOCUS       sequence-212851-       14241 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct to express dCas9 fused with an inactive Dnmt3a 
            catalytic domain (E664A, E756A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14241)
  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 14241)
  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_IM
FEATURES             Location/Qualifiers
     source          1..14241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        91..470
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        472..670
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     625..645
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             688..868
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    915..1040
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1533..1766
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1951..1995
                     /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    2293..2410
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2475..3686
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3523..3542
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(3737..3763,3764..3766,3767..6253,6254..6256,
                     6257..7840)
                     /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             7853..7873
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7877..7897
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7901..7921
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    8858..9446
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8911..8931)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9329..9340)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9449..9465)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9450..9466)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9971..10151
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10177..10194)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10183..10407
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10453..10881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10540..10559)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10750..10771
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10890..10910)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10934..11144
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        10956..11152
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      11003..11138
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11065..11084
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11200..11247
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11266..11640
                     /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    11770..11891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11807..11826)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11861..11880
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11940..11956)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11940..11956)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11953..11975)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11964..11980
                     /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(11988..11994,11995..12012,12013..12018))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12033..12054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12171..12188)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12342..12930)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12422..12441)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13101..13892,13893..13961))
                     /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     13724..13743
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13962..14066)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14141..14160)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
       61 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
      121 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      181 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      241 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      301 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      361 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      421 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      481 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      541 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      601 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      661 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
      721 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
      781 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
      841 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
      901 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
      961 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     1021 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     1081 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     1141 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     1201 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     1261 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     1321 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     1381 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     1441 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     1501 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     1561 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     1621 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     1681 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     1741 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     1801 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     1861 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     1921 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     1981 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     2041 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     2101 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     2161 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
     2221 tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac aaatggcagt
     2281 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     2341 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     2401 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa ttaacccgtg
     2461 tcggctccag atctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcct
     2521 cacggcgagc gctgccacgt cagacgaagg gcgcagcgag cgtcctgatc cttccgcccg
     2581 gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca
     2641 gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag
     2701 cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg
     2761 cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca
     2821 gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag
     2881 cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg
     2941 tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg
     3001 ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt
     3061 gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa
     3121 ggtcttgagg ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctggggcac
     3181 catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt
     3241 gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc
     3301 gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc
     3361 tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt
     3421 aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc
     3481 agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa
     3541 ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt
     3601 aatcatttgg gtcaatatgt aattttcagt gttagactag taaattgtcc gctaaattct
     3661 ggccgttttt ggcttttttg ttagacgaag cttgggctgc aggtcgactc tagaagttaa
     3721 cctcgagggc gcgcccatgg acaagaagta ttctatcgga ctggccatcg ggactaatag
     3781 cgtcgggtgg gccgtgatca ctgacgagta caaggtgccc tctaagaagt tcaaggtgct
     3841 cgggaacacc gaccggcatt ccatcaagaa aaatctgatc ggagctctcc tctttgattc
     3901 aggggagacc gctgaagcaa cccgcctcaa gcggactgct agacggcggt acaccaggag
     3961 gaagaaccgg atttgttacc ttcaagagat attctccaac gaaatggcaa aggtcgacga
     4021 cagcttcttc cataggctgg aagaatcatt cctcgtggaa gaggataaga agcatgaacg
     4081 gcatcccatc ttcggtaata tcgtcgacga ggtggcctat cacgagaaat acccaaccat
     4141 ctaccatctt cgcaaaaagc tggtggactc aaccgacaag gcagacctcc ggcttatcta
     4201 cctggccctg gcccacatga tcaagttcag aggccacttc ctgatcgagg gcgacctcaa
     4261 tcctgacaat agcgatgtgg ataaactgtt catccagctg gtgcagactt acaaccagct
     4321 ctttgaagag aaccccatca atgcaagcgg agtcgatgcc aaggccattc tgtcagcccg
     4381 gctgtcaaag agccgcagac ttgagaatct tatcgctcag ctgccgggtg aaaagaaaaa
     4441 tggactgttc gggaacctga ttgctctttc acttgggctg actcccaatt tcaagtctaa
     4501 tttcgacctg gcagaggatg ccaagctgca actgtccaag gacacctatg atgacgatct
     4561 cgacaacctc ctggcccaga tcggtgacca atacgccgac cttttccttg ctgctaagaa
     4621 tctttctgac gccatcctgc tgtctgacat tctccgcgtg aacactgaaa tcaccaaggc
     4681 ccctctttca gcttcaatga ttaagcggta tgatgagcac caccaggacc tgaccctgct
     4741 taaggcactc gtccggcagc agcttccgga gaagtacaag gaaatcttct ttgaccagtc
     4801 aaagaatgga tacgccggct acatcgacgg aggtgcctcc caagaggaat tttataagtt
     4861 tatcaaacct atccttgaga agatggacgg caccgaagag ctcctcgtga aactgaatcg
     4921 ggaggatctg ctgcggaagc agcgcacttt cgacaatggg agcattcccc accagatcca
     4981 tcttggggag cttcacgcca tccttcggcg ccaagaggac ttctacccct ttcttaagga
     5041 caacagggag aagattgaga aaattctcac tttccgcatc ccctactacg tgggacccct
     5101 cgccagagga aatagccggt ttgcttggat gaccagaaag tcagaagaaa ctatcactcc
     5161 ctggaacttc gaagaggtgg tggacaaggg agccagcgct cagtcattca tcgaacggat
     5221 gactaacttc gataagaacc tccccaatga gaaggtcctg ccgaaacatt ccctgctcta
     5281 cgagtacttt accgtgtaca acgagctgac caaggtgaaa tatgtcaccg aagggatgag
     5341 gaagcccgca ttcctgtcag gcgaacaaaa gaaggcaatt gtggaccttc tgttcaagac
     5401 caatagaaag gtgaccgtga agcagctgaa ggaggactat ttcaagaaaa ttgaatgctt
     5461 cgactctgtg gagattagcg gggtcgaaga tcggttcaac gcaagcctgg gtacctacca
     5521 tgatctgctt aagatcatca aggacaagga ttttctggac aatgaggaga acgaggacat
     5581 ccttgaggac attgtcctga ctctcactct gttcgaggac cgggaaatga tcgaggagag
     5641 gcttaagacc tacgcccatc tgttcgacga taaagtgatg aagcaactta aacggagaag
     5701 atataccgga tggggacgcc ttagccgcaa actcatcaac ggaatccggg acaaacagag
     5761 cggaaagacc attcttgatt tccttaagag cgacggattc gctaatcgca acttcatgca
     5821 acttatccat gatgattccc tgacctttaa ggaggacatc cagaaggccc aagtgtctgg
     5881 acaaggtgac tcactgcacg agcatatcgc aaatctggct ggttcacccg ctattaagaa
     5941 gggtattctc cagaccgtga aagtcgtgga cgagctggtc aaggtgatgg gtcgccataa
     6001 accagagaac attgtcatcg agatggccag ggaaaaccag actacccaga agggacagaa
     6061 gaacagcagg gagcggatga aaagaattga ggaagggatt aaggagctcg ggtcacagat
     6121 ccttaaagag cacccggtgg aaaacaccca gcttcagaat gagaagctct atctgtacta
     6181 ccttcaaaat ggacgcgata tgtatgtgga ccaagagctt gatatcaaca ggctctcaga
     6241 ctacgacgtg gacgccatcg tccctcagag cttcctcaaa gacgactcaa ttgacaataa
     6301 ggtgctgact cgctcagaca agaaccgggg aaagtcagat aacgtgccct cagaggaagt
     6361 cgtgaaaaag atgaagaact attggcgcca gcttctgaac gcaaagctga tcactcagcg
     6421 gaagttcgac aatctcacta aggctgagag gggcggactg agcgaactgg acaaagcagg
     6481 attcattaaa cggcaacttg tggagactcg gcagattact aaacatgtcg cccaaatcct
     6541 tgactcacgc atgaatacca agtacgacga aaacgacaaa cttatccgcg aggtgaaggt
     6601 gattaccctg aagtccaagc tggtcagcga tttcagaaag gactttcaat tctacaaagt
     6661 gcgggagatc aataactatc atcatgctca tgacgcatat ctgaatgccg tggtgggaac
     6721 cgccctgatc aagaagtacc caaagctgga aagcgagttc gtgtacggag actacaaggt
     6781 ctacgacgtg cgcaagatga ttgccaaatc tgagcaggag atcggaaagg ccaccgcaaa
     6841 gtacttcttc tacagcaaca tcatgaattt cttcaagacc gaaatcaccc ttgcaaacgg
     6901 tgagatccgg aagaggccgc tcatcgagac taatggggag actggcgaaa tcgtgtggga
     6961 caagggcaga gatttcgcta ccgtgcgcaa agtgctttct atgcctcaag tgaacatcgt
     7021 gaagaaaacc gaggtgcaaa ccggaggctt ttctaaggaa tcaatcctcc ccaagcgcaa
     7081 ctccgacaag ctcattgcaa ggaagaagga ttgggaccct aagaagtacg gcggattcga
     7141 ttcaccaact gtggcttatt ctgtcctggt cgtggctaag gtggaaaaag gaaagtctaa
     7201 gaagctcaag agcgtgaagg aactgctggg tatcaccatt atggagcgca gctccttcga
     7261 gaagaaccca attgactttc tcgaagccaa aggttacaag gaagtcaaga aggaccttat
     7321 catcaagctc ccaaagtata gcctgttcga actggagaat gggcggaagc ggatgctcgc
     7381 ctccgctggc gaacttcaga agggtaatga gctggctctc ccctccaagt acgtgaattt
     7441 cctctacctt gcaagccatt acgagaagct gaaggggagc cccgaggaca acgagcaaaa
     7501 gcaactgttt gtggagcagc ataagcatta tctggacgag atcattgagc agatttccga
     7561 gttttctaaa cgcgtcattc tcgctgatgc caacctcgat aaagtcctta gcgcatacaa
     7621 taagcacaga gacaaaccaa ttcgggagca ggctgagaat atcatccacc tgttcaccct
     7681 caccaatctt ggtgcccctg ccgcattcaa gtacttcgac accaccatcg accggaaacg
     7741 ctatacctcc accaaagaag tgctggacgc caccctcatc caccagagca tcaccggact
     7801 ttacgaaact cggattgacc tctcacagct cggaggggat gagggagctg atccaaaaaa
     7861 gaagagaaag gtagacccaa agaaaaaacg aaaagtagat ccgaaaaaga agaggaaggt
     7921 gggatccaac cacgaccagg aatttgaccc tccaaaggtt tacccacctg tcccagctga
     7981 gaagaggaag cccatccggg tgctgtctct ctttgatgga atcgctacag ggctcctggt
     8041 gctgaaggac ttgggcattc aggtggaccg ctacattgcc tcggccgtgt gtgaggactc
     8101 catcacggtg ggcatggtgc ggcaccaggg gaagatcatg tacgtcgggg acgtccgcag
     8161 cgtcacacag aagcatatcc aggagtgggg cccattcgat ctggtgattg ggggcagtcc
     8221 ctgcaatgac ctctccatcg tcaaccctgc tcgcaagggc ctctacgagg gcactggccg
     8281 gctcttcttt gagttctacc gcctcctgca tgatgcgcgg cccaaggagg gagatgatcg
     8341 ccccttcttc tggctctttg ccaatgtggt ggccatgggc gttagtgaca agagggacat
     8401 ctcgcgattt ctcgagtcca accctgtgat gattgatgcc aaagaagtgt cagctgcaca
     8461 cagggcccgc tacttctggg gtaaccttcc cggtatgaac aggccgttgg catccactgt
     8521 gaatgataag ctggagctgc aggagtgtct ggagcatggc aggatagcca agttcagcaa
     8581 agtgaggacc attactacga ggtcaaactc cataaagcag ggcaaagacc agcattttcc
     8641 tgtcttcatg aatgagaaag aggacatctt atggtgcact gaaatggaaa gggtatttgg
     8701 tttcccagtc cactatactg acgtctccaa catgagccgc ttggcgaggc agagactgct
     8761 gggccggtca tggagcgtgc cagtcatccg ccacctcttc gctccgctga aggagtattt
     8821 tgcgtgtgtg taagaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     8881 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     8941 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     9001 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     9061 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     9121 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     9181 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     9241 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     9301 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     9361 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     9421 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca
     9481 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
     9541 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
     9601 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     9661 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
     9721 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg
     9781 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga
     9841 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
     9901 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc
     9961 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
    10021 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
    10081 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
    10141 aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
    10201 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
    10261 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
    10321 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
    10381 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
    10441 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
    10501 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
    10561 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
    10621 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
    10681 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
    10741 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
    10801 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
    10861 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
    10921 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
    10981 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
    11041 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
    11101 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
    11161 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
    11221 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
    11281 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
    11341 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
    11401 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
    11461 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
    11521 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
    11581 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
    11641 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
    11701 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
    11761 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
    11821 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
    11881 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
    11941 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
    12001 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
    12061 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
    12121 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
    12181 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
    12241 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
    12301 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
    12361 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
    12421 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
    12481 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
    12541 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
    12601 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
    12661 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
    12721 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
    12781 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
    12841 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
    12901 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
    12961 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
    13021 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
    13081 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
    13141 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
    13201 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
    13261 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
    13321 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
    13381 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
    13441 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
    13501 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
    13561 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
    13621 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
    13681 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
    13741 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
    13801 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
    13861 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
    13921 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
    13981 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
    14041 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
    14101 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
    14161 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
    14221 gcaaaattta agctacaaca a
//
