LOCUS       sequence-213306-       13023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9 fused to BFP and the human DNMT3B catalytic domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13023)
  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 13023)
  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-DNMT3B
FEATURES             Location/Qualifiers
     source          1..13023
                     /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    8968..9556
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9021..9041)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9439..9450)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9645..9878
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(9907..9925)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(9908..9925)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(10171..10190)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10172..10190)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10200..10217)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10200..10216)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10209..10231)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10358..10813
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10445..10464)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10655..10676
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10839..10943
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10944..11012,11013..11804)
                     /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(11162..11181)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11975..12563
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12464..12483
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12717..12734
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12851..12872
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12887..12892,12893..12910,12911..12917)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12925..12941
                     /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     12930..12952
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12949..12965
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12949..12965
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12984..13004
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12986..13004
                     /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 agcgtcgacg tcctgcggcg ccggaaggac
     7981 tggaacgtgc gcctgcaggc cttcttcacc agtgacacgg ggcttgaata tgaagccccc
     8041 aagctgtacc ctgccattcc cgcagcccga aggcggccca ttcgagtcct gtcattgttt
     8101 gatggcatcg cgacaggcta cctagtcctc aaagagttgg gcataaaggt aggaaagtac
     8161 gtcgcttctg aagtgtgtga ggagtccatt gctgttggaa ccgtgaagca cgaggggaat
     8221 atcaaatacg tgaacgacgt gaggaacatc acaaagaaaa atattgaaga atggggccca
     8281 tttgacttgg tgattggcgg aagcccatgc aacgatctct caaatgtgaa tccagccagg
     8341 aaaggcctgt atgagggtac aggccggctc ttcttcgaat tttaccacct gctgaattac
     8401 tcacgcccca aggagggtga tgaccggccg ttcttctgga tgtttgagaa tgttgtagcc
     8461 atgaaggttg gcgacaagag ggacatctca cggttcctgg agtgtaatcc agtgatgatt
     8521 gatgccatca aagtttctgc tgctcacagg gcccgatact tctggggcaa cctacccggg
     8581 atgaacaggc ccgtgatagc atcaaagaat gataaactcg agctgcagga ctgcttggaa
     8641 tacaatagga tagccaagtt aaagaaagta cagacaataa ccaccaagtc gaactcgatc
     8701 aaacagggga aaaaccaact tttccctgtt gtcatgaatg gcaaagaaga tgttttgtgg
     8761 tgcactgagc tcgaaaggat ctttggcttt cctgtgcact acacagacgt gtccaacatg
     8821 ggccgtggtg cccgccagaa gctgctggga aggtcctgga gcgtgcctgt catccgacac
     8881 ctcttcgccc ctctgaagga ctactttgca tgtgaatagt aaatgcatgg ccggccttaa
     8941 ttaaacgcgt ctcgagggaa ttccgataat caacctctgg attacaaaat ttgtgaaaga
     9001 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     9061 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     9121 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     9181 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     9241 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     9301 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     9361 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     9421 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     9481 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     9541 tgggccgcct ccccgcatcg ggaattcgag ctcggtacct ttaagaccaa tgacttacaa
     9601 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     9661 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     9721 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     9781 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     9841 cctcagaccc ttttagtcag tgtggaaaat ctctagcagc atctagctag aattaattcc
     9901 gtgtattcta tagtgtcacc taaatcgtat gtgtatgata cataaggtta tgtattaatt
     9961 gtagccgcgt tctaacgaca atatgtacaa gcctaattgt gtagcatctg gcttactgaa
    10021 gcagacccta tcatctctct cgtaaactgc cgtcagagtc ggtttggttg gacgaacctt
    10081 ctgagtttct ggtaacgccg tcccgcaccc ggaaatggtc agcgaaccaa tcagcagggt
    10141 catcgctagc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
    10201 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
    10261 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
    10321 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
    10381 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
    10441 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
    10501 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
    10561 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
    10621 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
    10681 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
    10741 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
    10801 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
    10861 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
    10921 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
    10981 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
    11041 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
    11101 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
    11161 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
    11221 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
    11281 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
    11341 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
    11401 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
    11461 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
    11521 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
    11581 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
    11641 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
    11701 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
    11761 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
    11821 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
    11881 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
    11941 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
    12001 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
    12061 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
    12121 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
    12181 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
    12241 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
    12301 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
    12361 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
    12421 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
    12481 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
    12541 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
    12601 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
    12661 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
    12721 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
    12781 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
    12841 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
    12901 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
    12961 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
    13021 tgc
//
