LOCUS       sequence-213289-       12969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9 fused to BFP and the human DNMT3A catalytic domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12969)
  AUTHORS   Lin L, Liu Y, Xu F, Huang J, Daugaard TF, Petersen TS, Hansen B, Ye 
            L, Zhou Q, Fang F, Yang L, Li S, Floe L, Jensen KT, Shrock E, Chen 
            F, Yang H, Wang J, Liu X, Xu X, Bolund L, Nielsen AL, Luo Y
  TITLE     Genome-wide determination of on-target and off-target 
            characteristics for RNA-guided DNA methylation by dCas9 
            methyltransferases.
  JOURNAL   Gigascience. 2018 Mar 1;7(3):1-19. doi: 10.1093/gigascience/giy011.
  PUBMED    29635374
REFERENCE   2  (bases 1 to 12969)
  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"; journalName: 
            "Gigascience"; date: "2018-03-1- 1"; volume: "7"; issue: "3"; pages:
            "1-19"
COMMENT     Sequence Label: pCCL-PGK-SPdCas9-BFP-DNMT3A
FEATURES             Location/Qualifiers
     source          1..12969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        82..461
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        462..665
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     616..636
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             680..860
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    907..1032
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1525..1758
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1943..1987
                     /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    2254..2371
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2410..2909
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2445..2468)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2842..2861
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      2968..2977
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2974..3000,3001..3003,3004..5490,5491..5493,
                     5494..7077)
                     /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             2974..7077
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /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             7081..7107
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7087..7113
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             7126..7146
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7153..7173
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7213..7908
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     misc_feature    8914..9502
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8967..8987)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9385..9396)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9591..9824
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(9853..9871)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(9854..9871)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(10117..10136)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10118..10136)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10146..10163)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10146..10162)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10155..10177)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10304..10759
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10391..10410)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10601..10622
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10785..10889
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10890..10958,10959..11750)
                     /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     complement(11108..11127)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11921..12509
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12410..12429
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12663..12680
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12797..12818
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12833..12838,12839..12856,12857..12863)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12871..12887
                     /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)."
     primer_bind     12876..12898
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12895..12911
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12895..12911
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12930..12950
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12932..12950
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aagcttggcc attgcatacg ttgtatccat atcataatat gtacatttat attggctcat
       61 gtccaacatt accgccatgt tgacattgat tattgactag ttattaatag taatcaatta
      121 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      181 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      241 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      301 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      361 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      421 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      481 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      541 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      601 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      661 gagctcgttt agtgaaccgg ggtctctctg gttagaccag atctgagcct gggagctctc
      721 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
      781 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
      841 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
      901 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
      961 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     1021 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     1081 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     1141 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     1201 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     1261 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     1321 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
     1381 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     1441 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     1501 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     1561 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     1621 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     1681 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     1741 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     1801 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     1861 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     1921 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     1981 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     2041 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     2101 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     2161 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     2221 attagtgaac ggatctcgac ggtatcggtt aacttttaaa agaaaagggg ggattggggg
     2281 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     2341 acaaaaacaa attacaaaaa ttcaaaattt tatcgatcac gagactagcc tcgacgatgg
     2401 tcgagtaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag
     2461 ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca
     2521 ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc
     2581 ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg
     2641 aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg
     2701 taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc
     2761 tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc
     2821 gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt
     2881 tctcctcttc ctcatctccg ggcctttcga cctctagcgg gatccggcgc gtctgcagga
     2941 tatcaagctt gcggtaccgc gggcccggcc accatggaca agaagtacag catcggcctg
     3001 gccatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc
     3061 aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcggc
     3121 gccctgctgt tcgacagcgg agaaacagcc gaggccaccc ggctgaagag aaccgccaga
     3181 agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag
     3241 atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag
     3301 gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac
     3361 gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc
     3421 gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg
     3481 atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg
     3541 cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag
     3601 gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg
     3661 cccggcgaga agaagaatgg cctgttcggc aacctgattg ccctgagcct gggcctgacc
     3721 cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac
     3781 acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg
     3841 tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac
     3901 accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac
     3961 caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag
     4021 attttcttcg accagagcaa gaacggctac gccggctaca tcgatggcgg agccagccag
     4081 gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg
     4141 ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc
     4201 atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt
     4261 tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc
     4321 tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc
     4381 gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc cagcgcccag
     4441 agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc
     4501 aagcacagcc tgctgtacga gtacttcacc gtgtacaacg agctgaccaa agtgaaatac
     4561 gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa agccatcgtg
     4621 gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc
     4681 aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc
     4741 tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat
     4801 gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga
     4861 gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag
     4921 cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc
     4981 atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc
     5041 aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag
     5101 aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc
     5161 agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa
     5221 gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc
     5281 acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa
     5341 gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag
     5401 aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac
     5461 atcaaccggc tgtccgacta cgatgtggac gctatcgtgc ctcagagctt tctgaaggac
     5521 gactccatcg ataacaaagt gctgactcgg agcgacaaga accggggcaa gagcgacaac
     5581 gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcgccagct gctgaatgcc
     5641 aagctgatta cccagaggaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc
     5701 gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag
     5761 cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa cgacaaactg
     5821 atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat
     5881 ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg
     5941 aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg
     6001 tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc
     6061 ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag
     6121 attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaaca
     6181 ggcgagatcg tgtgggataa gggccgggac tttgccaccg tgcggaaagt gctgtctatg
     6241 ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct
     6301 atcctgccca agaggaacag cgacaagctg atcgccagaa agaaggactg ggaccctaag
     6361 aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg
     6421 gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg
     6481 gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa
     6541 gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc
     6601 cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc
     6661 tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc
     6721 gaggataatg agcagaaaca gctgtttgtg gaacagcaca aacactacct ggacgagatc
     6781 atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaag
     6841 gtgctgagcg cctacaacaa gcacagagac aagcctatca gagagcaggc cgagaatatc
     6901 atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc
     6961 accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac
     7021 cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacgcc
     7081 tatccctatg acgtgcccga ttatgccagc ctgggcagcg gctcccccaa gaaaaaacgc
     7141 aaggtggaag atcctaagaa aaagcggaaa gtggacggca ttggtagtgg gagcaacggc
     7201 agcagcggat ccagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc
     7261 accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc
     7321 acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc
     7381 ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc
     7441 gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa
     7501 gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac
     7561 aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca
     7621 ctcggctggg aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga
     7681 aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacat caagaccaca
     7741 tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac
     7801 agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca
     7861 gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaatgg tggaggaggt
     7921 tctggaggcg gtggaagtgg tggcggaggt agcgtcgaca accacgacca ggaatttgac
     7981 cctccaaagg tttacccacc tgtcccagct gagaagagga agcccatccg ggtgctgtct
     8041 ctctttgatg gaatcgctac agggctcctg gtgctgaagg acttgggcat tcaggtggac
     8101 cgctacattg cctcggaggt gtgtgaggac tccatcacgg tgggcatggt gcggcaccag
     8161 gggaagatca tgtacgtcgg ggacgtccgc agcgtcacac agaagcatat ccaggagtgg
     8221 ggcccattcg atctggtgat tgggggcagt ccctgcaatg acctctccat cgtcaaccct
     8281 gctcgcaagg gcctctacga gggcactggc cggctcttct ttgagttcta ccgcctcctg
     8341 catgatgcgc ggcccaagga gggagatgat cgccccttct tctggctctt tgagaatgtg
     8401 gtggccatgg gcgttagtga caagagggac atctcgcgat ttctcgagtc caaccctgtg
     8461 atgattgatg ccaaagaagt gtcagctgca cacagggccc gctacttctg gggtaacctt
     8521 cccggtatga acaggccgtt ggcatccact gtgaatgata agctggagct gcaggagtgt
     8581 ctggagcatg gcaggatagc caagttcagc aaagtgagga ccattactac gaggtcaaac
     8641 tccataaagc agggcaaaga ccagcatttt cctgtcttca tgaatgagaa agaggacatc
     8701 ttatggtgca ctgaaatgga aagggtattt ggtttcccag tccactatac tgacgtctcc
     8761 aacatgagcc gcttggcgag gcagagactg ctgggccggt catggagcgt gccagtcatc
     8821 cgccacctct tcgctccgct gaaggagtat tttgcgtgtg tgtaatagat gcatggccgg
     8881 ccttaattaa acgcgtctcg agggaattcc gataatcaac ctctggatta caaaatttgt
     8941 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     9001 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     9061 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     9121 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     9181 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     9241 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     9301 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     9361 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     9421 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     9481 tccctttggg ccgcctcccc gcatcgggaa ttcgagctcg gtacctttaa gaccaatgac
     9541 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     9601 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
     9661 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     9721 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     9781 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagcatct agctagaatt
     9841 aattccgtgt attctatagt gtcacctaaa tcgtatgtgt atgatacata aggttatgta
     9901 ttaattgtag ccgcgttcta acgacaatat gtacaagcct aattgtgtag catctggctt
     9961 actgaagcag accctatcat ctctctcgta aactgccgtc agagtcggtt tggttggacg
    10021 aaccttctga gtttctggta acgccgtccc gcacccggaa atggtcagcg aaccaatcag
    10081 cagggtcatc gctagcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg
    10141 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
    10201 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
    10261 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
    10321 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
    10381 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
    10441 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
    10501 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
    10561 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
    10621 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
    10681 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
    10741 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
    10801 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
    10861 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
    10921 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
    10981 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
    11041 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
    11101 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
    11161 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
    11221 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
    11281 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
    11341 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
    11401 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
    11461 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
    11521 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
    11581 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
    11641 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
    11701 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
    11761 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
    11821 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
    11881 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
    11941 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
    12001 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
    12061 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
    12121 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
    12181 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
    12241 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
    12301 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
    12361 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
    12421 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
    12481 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
    12541 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
    12601 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
    12661 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
    12721 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
    12781 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
    12841 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
    12901 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
    12961 tggagctgc
//
