LOCUS       sequence-345683-       12028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the dCas9-DNMT3a-DNMT3l-3xFLAG-tagBFP fusion protein for 
            targeted DNA methylation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12028)
  AUTHORS   Hofacker D, Broche J, Laistner L, Adam S, Bashtrykov P, Jeltsch A
  TITLE     Engineering of Effector Domains for Targeted DNA Methylation with 
            Reduced Off-Target Effects.
  JOURNAL   Int J Mol Sci. 2020 Jan 13;21(2). pii: ijms21020502. doi: 
            10.3390/ijms21020502.
  PUBMED    31941101
REFERENCE   2  (bases 1 to 12028)
  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: "Int J Mol 
            Sci."; date: "2020-01-13"; volume: "21(2). pii: ijms21020502. doi"; 
            pages: " 10.3390/ijms21020502"
COMMENT     Sequence Label: dCas9-DNMT3a-DNMT3l-3xFLAG-tagBFP
FEATURES             Location/Qualifiers
     source          1..12028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(180..199)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        371..750
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        751..954
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     905..925
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     951..975
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1063..1072
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1069..1095,1096..1098,1099..3582,3583..3588,3589..3654,
                     3655..3657,3658..5172)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5185..5205
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6886..6906
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6907..6972
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6982..7683
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(7735..7754)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7779..7827
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8029..8457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8116..8135)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8326..8347
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8466..8486)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8471..8800
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8651..8786
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8713..8732
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8867..9661
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8921..8940)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9531..9550
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9837..9958
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9874..9893)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9928..9947
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10007..10023)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10007..10023)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10020..10042)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10031..10047
                     /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(10055..10061,10062..10079,10080..10085))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10100..10121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10238..10255)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10409..10997)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10489..10508)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11168..11959,11960..12028))
                     /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     11791..11810
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
      181 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
      241 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
      301 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
      361 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      421 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      481 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      541 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      601 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      661 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      721 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      781 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      841 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      901 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      961 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     1021 cgggaccgat ccagcctccg gactctagag gatcgaaccc ttgccaccat ggacaagaag
     1081 tactccattg ggctcgctat cggcacaaac agcgtcggct gggccgtcat tacggacgag
     1141 tacaaggtgc cgagcaaaaa attcaaagtt ctgggcaata ccgatcgcca cagcataaag
     1201 aagaacctca ttggcgccct cctgttcgac tccggggaga cggccgaagc cacgcggctc
     1261 aaaagaacag cacggcgcag atatacccgc agaaagaatc ggatctgcta cctgcaggag
     1321 atctttagta atgagatggc taaggtggat gactctttct tccataggct ggaggagtcc
     1381 tttttggtgg aggaggataa aaagcacgag cgccacccaa tctttggcaa tatcgtggac
     1441 gaggtggcgt accatgaaaa gtacccaacc atatatcatc tgaggaagaa gcttgtagac
     1501 agtactgata aggctgactt gcggttgatc tatctcgcgc tggcgcatat gatcaaattt
     1561 cggggacact tcctcatcga gggggacctg aacccagaca acagcgatgt cgacaaactc
     1621 tttatccaac tggttcagac ttacaatcag cttttcgaag agaacccgat caacgcatcc
     1681 ggagttgacg ccaaagcaat cctgagcgct aggctgtcca aatcccggcg gctcgaaaac
     1741 ctcatcgcac agctccctgg ggagaagaag aacggcctgt ttggtaatct tatcgccctg
     1801 tcactcgggc tgacccccaa ctttaaatct aacttcgacc tggccgaaga tgccaagctt
     1861 caactgagca aagacaccta cgatgatgat ctcgacaatc tgctggccca gatcggcgac
     1921 cagtacgcag accttttttt ggcggcaaag aacctgtcag acgccattct gctgagtgat
     1981 attctgcgag tgaacacgga gatcaccaaa gctccgctga gcgctagtat gatcaagcgc
     2041 tatgatgagc accaccaaga cttgactttg ctgaaggccc ttgtcagaca gcaactgcct
     2101 gagaagtaca aggaaatttt cttcgatcag tctaaaaatg gctacgccgg atacattgac
     2161 ggcggagcaa gccaggagga attttacaaa tttattaagc ccatcttgga aaaaatggac
     2221 ggcaccgagg agctgctggt aaagcttaac agagaagatc tgttgcgcaa acagcgcact
     2281 ttcgacaatg gaagcatccc ccaccagatt cacctgggcg aactgcacgc tatcctcagg
     2341 cggcaagagg atttctaccc ctttttgaaa gataacaggg aaaagattga gaaaatcctc
     2401 acatttcgga taccctacta tgtaggcccc ctcgcccggg gaaattccag attcgcgtgg
     2461 atgactcgca aatcagaaga gaccatcact ccctggaact tcgaggaagt cgtggataag
     2521 ggggcctctg cccagtcctt catcgaaagg atgactaact ttgataaaaa tctgcctaac
     2581 gaaaaggtgc ttcctaaaca ctctctgctg tacgagtact tcacagttta taacgagctc
     2641 accaaggtca aatacgtcac agaagggatg agaaagccag cattcctgtc tggagagcag
     2701 aagaaagcta tcgtggacct cctcttcaag acgaaccgga aagttaccgt gaaacagctc
     2761 aaagaagact atttcaaaaa gattgaatgt ttcgactctg ttgaaatcag cggagtggag
     2821 gatcgcttca acgcatccct gggaacgtat cacgatctcc tgaaaatcat taaagacaag
     2881 gacttcctgg acaatgagga gaacgaggac attcttgagg acattgtcct cacccttacg
     2941 ttgtttgaag atagggagat gattgaagaa cgcttgaaaa cttacgctca tctcttcgac
     3001 gacaaagtca tgaaacagct caagaggcgc cgatatacag gatgggggcg gctgtcaaga
     3061 aaactgatca atgggatccg agacaagcag agtggaaaga caatcctgga ttttcttaag
     3121 tccgatggat ttgccaaccg gaacttcatg cagttgatcc atgatgactc tctcaccttt
     3181 aaggaggaca tccagaaagc acaagtttct ggccaggggg acagtcttca cgagcacatc
     3241 gctaatcttg caggtagccc agctatcaaa aagggaatac tgcagaccgt taaggtcgtg
     3301 gatgaactcg tcaaagtaat gggaaggcat aagcccgaga atatcgttat cgagatggcc
     3361 cgagagaacc aaactaccca gaagggacag aagaacagta gggaaaggat gaagaggatt
     3421 gaagagggta taaaagaact ggggtcccaa atccttaagg aacacccagt tgaaaacacc
     3481 cagcttcaga atgagaagct ctacctgtac tacctgcaga acggcaggga catgtacgtg
     3541 gatcaggaac tggacatcaa tcggctctcc gactacgacg tggctgctat cgtgccccag
     3601 tcttttctca aagatgattc tattgataat aaagtgttga caagatccga taaagctaga
     3661 gggaagagtg ataacgtccc ctcagaagaa gttgtcaaga aaatgaaaaa ttattggcgg
     3721 cagctgctga acgccaaact gatcacacaa cggaagttcg ataatctgac taaggctgaa
     3781 cgaggtggcc tgtctgagtt ggataaagcc ggcttcatca aaaggcagct tgttgagaca
     3841 cgccagatca ccaagcacgt ggcccaaatt ctcgattcac gcatgaacac caagtacgat
     3901 gaaaatgaca aactgattcg agaggtgaaa gttattactc tgaagtctaa gctggtctca
     3961 gatttcagaa aggactttca gttttataag gtgagagaga tcaacaatta ccaccatgcg
     4021 catgatgcct acctgaatgc agtggtaggc actgcactta tcaaaaaata tcccaagctt
     4081 gaatctgaat ttgtttacgg agactataaa gtgtacgatg ttaggaaaat gatcgcaaag
     4141 tctgagcagg aaataggcaa ggccaccgct aagtacttct tttacagcaa tattatgaat
     4201 tttttcaaga ccgagattac actggccaat ggagagattc ggaagcgacc acttatcgaa
     4261 acaaacggag aaacaggaga aatcgtgtgg gacaagggta gggatttcgc gacagtccgg
     4321 aaggtcctgt ccatgccgca ggtgaacatc gttaaaaaga ccgaagtaca gaccggaggc
     4381 ttctccaagg aaagtatcct cccgaaaagg aacagcgaca agctgatcgc acgcaaaaaa
     4441 gattgggacc ccaagaaata cggcggattc gattctccta cagtcgctta cagtgtactg
     4501 gttgtggcca aagtggagaa agggaagtct aaaaaactca aaagcgtcaa ggaactgctg
     4561 ggcatcacaa tcatggagcg atcaagcttc gaaaaaaacc ccatcgactt tctcgaggcg
     4621 aaaggatata aagaggtcaa aaaagacctc atcattaagc ttcccaagta ctctctcttt
     4681 gagcttgaaa acggccggaa acgaatgctc gctagtgcgg gcgagctgca gaaaggtaac
     4741 gagctggcac tgccctctaa atacgttaat ttcttgtatc tggccagcca ctatgaaaag
     4801 ctcaaagggt ctcccgaaga taatgagcag aagcagctgt tcgtggaaca acacaaacac
     4861 taccttgatg agatcatcga gcaaataagc gaattctcca aaagagtgat cctcgccgac
     4921 gctaacctcg ataaggtgct ttctgcttac aataagcaca gggataagcc catcagggag
     4981 caggcagaaa acattatcca cttgtttact ctgaccaact tgggcgcgcc tgcagccttc
     5041 aagtacttcg acaccaccat agacagaaag cggtacacct ctacaaagga ggtcctggac
     5101 gccacactga ttcatcagtc aattacgggg ctctatgaaa caagaatcga cctctctcag
     5161 ctcggtggag acagcagggc tgaccccaag aagaagagga aggtggaggc cagcggttcc
     5221 tccggactca gatctcgagc tcaaaaccat gaccaggaat ttgacccccc aaaggtttac
     5281 ccacctgtgc cagctgagaa gaggaagccc atccgcgtgc tgtctctctt tgatgggatt
     5341 gctacagggc tcctggtgct gaaggacctg ggcatccaag tggaccgcta cattgcctcc
     5401 gaggtgtgtg aggactccat cacggtgggc atggtgcggc accagggaaa gatcatgtac
     5461 gtcggggacg tccgcagcgt cacacagaag catatccagg agtggggccc attcgacctg
     5521 gtgattggag gcagtccctg caatgacctc tccattgtca accctgcccg caagggactt
     5581 tatgagggta ctggccgcct cttctttgag ttctaccgcc tcctgcatga tgcgcggccc
     5641 aaggagggag atgatcgccc cttcttctgg ctctttgaga atgtggtggc catgggcgtt
     5701 agtgacaaga gggacatctc gcgatttctt gagtctaacc ccgtgatgat tgacgccaaa
     5761 gaagtgtctg ctgcacacag ggcccgttac ttctggggta accttcctgg catgaacagg
     5821 cctttggcat ccactgtgaa tgataagctg gagctgcaag agtgtctgga gcacggcaga
     5881 atagccaagt tcagcaaagt gaggaccatt accaccaggt caaactctat aaagcagggc
     5941 aaagaccagc atttccccgt cttcatgaac gagaaggagg acatcctgtg gtgcactgaa
     6001 atggaaaggg tgtttggctt ccccgtccac tacacagacg tctccaacat gagccgcttg
     6061 gcgaggcaga gactgctggg ccgatcgtgg agcgtgccgg tcatccgcca cctcttcgct
     6121 ccgctgaagg aatattttgc ttgtgtgtct agcggcaata gtaacgctaa cagccgcggg
     6181 ccgagcttca gcagcggcct ggtgccgtta agcttgcgcg gcagccatat gggccctatg
     6241 gagatataca agacagtgtc tgcatggaag agacagccag tgcgggtact gagccttttt
     6301 agaaatattg ataaagtact aaagagtttg ggctttttag aaagcggttc tggttctggg
     6361 ggaggaacgc tgaagtacgt ggaagatgtc acaaatgtcg tgaggagaga cgtggagaaa
     6421 tggggcccct ttgacctggt gtacggctcg acgcagcccc taggcagctc ctgtgatcgc
     6481 tgtcccggct ggtacatgtt ccagttccac cggatcctgc agtatgcgct gcctcgccag
     6541 gagagtcagc ggcccttctt ctggatattc atggacaatc tgctgctgac tgaggatgac
     6601 caagagacaa ctacccgctt ccttcagaca gaggctgtga ccctccagga tgtccgtggc
     6661 agggactacc agaatgctat gcgggtgtgg agcaacattc cagggctgaa gagcaagcat
     6721 gcgcccctga ccccaaagga agaagagtat ctgcaagccc aagtcagaag caggagcaag
     6781 ctggacgccc cgaaagttga cctcctggtg aagaactgcc ttctcccgct gagagagtac
     6841 ttcaagtatt tttctcaaaa ctcacttcct cttagttccg gatctccgaa aaagaaacgc
     6901 aaagttgact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
     6961 gacgatgaca aggtcgccac catgagcgag ctgattaagg agaacatgca catgaagctg
     7021 tacatggagg gcaccgtgga caaccatcac ttcaagtgca catccgaggg cgaaggcaag
     7081 ccctacgagg gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc
     7141 gccttcgaca tcctggctac tagcttcctc tacggcagca agaccttcat caaccacacc
     7201 cagggcatcc ccgacttctt caagcagtcc ttccctgagg gcttcacatg ggagagagtc
     7261 accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc
     7321 tgcctcatct acaacgtcaa gatcagaggg gtgaacttca catccaacgg ccctgtgatg
     7381 cagaagaaaa cactcggctg ggaggccttc accgagacgc tgtaccccgc tgacggcggc
     7441 ctggaaggca gaaacgacat ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac
     7501 atcaagacca catatagatc caagaaaccc gctaagaacc tcaagatgcc tggcgtctac
     7561 tatgtggact acagactgga aagaatcaag gaggccaaca acgagaccta cgtcgagcag
     7621 cacgaggtgg cagtggccag atactgcgac ctccctagca aactggggca caagcttaat
     7681 tgaaagggtt cgatccctac cggttagtaa tgagtttaaa cgggggaggc taactgaaac
     7741 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     7801 aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     7861 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     7921 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     7981 gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta
     8041 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     8101 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     8161 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     8221 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     8281 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     8341 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     8401 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     8461 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     8521 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     8581 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     8641 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     8701 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     8761 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     8821 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     8881 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     8941 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     9001 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     9061 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     9121 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     9181 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     9241 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     9301 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     9361 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     9421 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     9481 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     9541 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     9601 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     9661 agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     9721 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     9781 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     9841 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     9901 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     9961 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
    10021 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
    10081 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
    10141 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
    10201 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
    10261 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
    10321 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
    10381 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
    10441 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
    10501 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
    10561 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
    10621 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
    10681 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
    10741 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
    10801 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
    10861 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
    10921 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
    10981 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
    11041 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
    11101 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
    11161 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
    11221 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
    11281 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
    11341 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
    11401 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
    11461 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
    11521 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
    11581 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
    11641 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
    11701 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
    11761 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
    11821 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
    11881 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
    11941 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
    12001 gggcgacacg gaaatgttga atactcat
//
