LOCUS       sequence-206110-       10930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BPF version of LLP339, Puro is replaced by BFP driven by EF1a, dCas9
            with SungTagx10 driven by pGK, with 3xHA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10930)
  AUTHORS   Pflueger C, Tan D, Swain T, Nguyen TV, Pflueger J, Nefzger C, Polo 
            JM, Ford E, Lister R
  TITLE     A modular dCas9-SunTag DNMT3A epigenome editing system overcomes 
            pervasive off-target activity of direct fusion dCas9-DNMT3A 
            constructs.
  JOURNAL   Genome Res. 2018 Jun 15. pii: gr.233049.117. doi: 
            10.1101/gr.233049.117.
  PUBMED    29907613
REFERENCE   2  (bases 1 to 10930)
  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: "Genome Res.
            2018 Jun 15. pii: gr.233049.117. doi: 10.1101/gr.233049.117."
COMMENT     Sequence Label: LLP457 pGK-dCas9-Suntag-BFP
FEATURES             Location/Qualifiers
     source          1..10930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        213..712
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(248..271)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     645..664
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             744..764
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             765..791
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             795..821
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             825..851
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(885..911,912..914,915..3398,3399..3404,3405..3470,
                     3471..3473,3474..4988)
                     /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
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(885..911,912..914,915..3401,3402..3404,3405..4988)
                     /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             885..4988
                     /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             5025..5045
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5085..5105
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5127..5183
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5199..5255
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5271..5327
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5343..5399
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5415..5471
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5487..5543
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5559..5615
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5631..5687
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5703..5759
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5775..5831
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5856..5876
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5896..5915)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5940..5987
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        6233..7411
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6464..7402
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7359..7379
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      7418..7427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7424..8134
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     intron          8176..8405
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     8388..8407
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     polyA_signal    8550..8671
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8587..8606)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(8773..8789)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8773..8789)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8786..8808)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8797..8813
                     /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(8821..8827,8828..8845,8846..8851))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8866..8887
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9004..9021)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9175..9763)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9255..9274)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9934..10725,10726..10794))
                     /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     10557..10576
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10795..10899)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ttaattaata ccgggtaggg gaggcgcttt tcccaaggca
      241 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
      301 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
      361 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
      421 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
      481 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
      541 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg
      601 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
      661 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cggcggccgc
      721 acctgcagcc caagcttacc atgccaaaga agaagcggaa ggtctacccc tatgacgtgc
      781 ctgattacgc cgcttatcct tacgatgtgc ccgactatgc agcctaccca tacgatgtgc
      841 ccgactacgc cgcaatggcc ggtatccacg gagtcccagc agccatggac aagaagtact
      901 ccattgggct cgctatcggc acaaacagcg tcggctgggc cgtcattacg gacgagtaca
      961 aggtgccgag caaaaaattc aaagttctgg gcaataccga tcgccacagc ataaagaaga
     1021 acctcattgg cgccctcctg ttcgactccg gggagacggc cgaagccacg cggctcaaaa
     1081 gaacagcacg gcgcagatat acccgcagaa agaatcggat ctgctacctg caggagatct
     1141 ttagtaatga gatggctaag gtggatgact ctttcttcca taggctggag gagtcctttt
     1201 tggtggagga ggataaaaag cacgagcgcc acccaatctt tggcaatatc gtggacgagg
     1261 tggcgtacca tgaaaagtac ccaaccatat atcatctgag gaagaagctt gtagacagta
     1321 ctgataaggc tgacttgcgg ttgatctatc tcgcgctggc gcatatgatc aaatttcggg
     1381 gacacttcct catcgagggg gacctgaacc cagacaacag cgatgtcgac aaactcttta
     1441 tccaactggt tcagacttac aatcagcttt tcgaagagaa cccgatcaac gcatccggag
     1501 ttgacgccaa agcaatcctg agcgctaggc tgtccaaatc ccggcggctc gaaaacctca
     1561 tcgcacagct ccctggggag aagaagaacg gcctgtttgg taatcttatc gccctgtcac
     1621 tcgggctgac ccccaacttt aaatctaact tcgacctggc cgaagatgcc aagcttcaac
     1681 tgagcaaaga cacctacgat gatgatctcg acaatctgct ggcccagatc ggcgaccagt
     1741 acgcagacct ttttttggcg gcaaagaacc tgtcagacgc cattctgctg agtgatattc
     1801 tgcgagtgaa cacggagatc accaaagctc cgctgagcgc tagtatgatc aagcgctatg
     1861 atgagcacca ccaagacttg actttgctga aggcccttgt cagacagcaa ctgcctgaga
     1921 agtacaagga aattttcttc gatcagtcta aaaatggcta cgccggatac attgacggcg
     1981 gagcaagcca ggaggaattt tacaaattta ttaagcccat cttggaaaaa atggacggca
     2041 ccgaggagct gctggtaaag cttaacagag aagatctgtt gcgcaaacag cgcactttcg
     2101 acaatggaag catcccccac cagattcacc tgggcgaact gcacgctatc ctcaggcggc
     2161 aagaggattt ctaccccttt ttgaaagata acagggaaaa gattgagaaa atcctcacat
     2221 ttcggatacc ctactatgtg ggccccctcg cccggggaaa ttccagattc gcgtggatga
     2281 ctcgcaaatc agaagagacc atcactccct ggaacttcga ggaagtcgtg gataaggggg
     2341 cctctgccca gtccttcatc gaaaggatga ctaactttga taaaaatctg cctaacgaaa
     2401 aggtgcttcc taaacactct ctgctgtacg agtacttcac agtttataac gagctcacca
     2461 aggtcaaata cgtcacagaa gggatgagaa agccagcatt cctgtctgga gagcagaaga
     2521 aagctatcgt ggacctcctc ttcaagacga accggaaagt taccgtgaaa cagctcaaag
     2581 aagactattt caaaaagatt gaatgtttcg actctgttga aatcagcgga gtggaggatc
     2641 gcttcaacgc atccctggga acgtatcacg atctcctgaa aatcattaaa gacaaggact
     2701 tcctggacaa tgaggagaac gaggacattc ttgaggacat tgtcctcacc cttacgttgt
     2761 ttgaagatag ggagatgatt gaagaacgct tgaaaactta cgctcatctc ttcgacgaca
     2821 aagtcatgaa acagctcaag aggcgccgat atacaggatg ggggcggctg tcaagaaaac
     2881 tgatcaatgg gatccgagac aagcagagtg gaaagacaat cctggatttt cttaagtccg
     2941 atggatttgc caacaggaac ttcatgcagt tgatccatga tgactctctc acctttaagg
     3001 aggacatcca gaaagcacaa gtttctggcc agggggacag tcttcacgag cacatcgcta
     3061 atcttgcagg tagcccagct atcaaaaagg gaatactgca gaccgttaag gtcgtggatg
     3121 aactcgtcaa agtaatggga aggcataagc ccgagaatat cgttatcgag atggcccgag
     3181 agaaccaaac tacccagaag ggacagaaga acagtaggga aaggatgaag aggattgaag
     3241 agggtataaa agaactgggg tcccaaatcc ttaaggaaca cccagttgaa aacacccagc
     3301 ttcagaatga gaagctctac ctgtactacc tgcagaacgg cagggacatg tacgtggatc
     3361 aggaactgga catcaatcgg ctctccgact acgacgtgga tgcaatcgtg ccccagtctt
     3421 ttctcaaaga tgattctatt gataataaag tgttgacaag atccgataaa aatagaggga
     3481 agagtgataa cgtcccctca gaagaagttg tcaagaaaat gaaaaattat tggcggcagc
     3541 tgctgaacgc caaactgatc acacaacgga agttcgataa tctgactaag gctgaacgag
     3601 gtggcctgtc tgagttggat aaagccggct tcatcaaaag gcagcttgtt gagacacgcc
     3661 agatcaccaa gcacgtggcc caaattctcg attcacgcat gaacaccaag tacgatgaaa
     3721 atgacaaact gattcgagag gtgaaagtta ttactctgaa gtctaagctg gtctcagatt
     3781 tcagaaagga ctttcagttt tataaggtga gagagatcaa caattaccac catgcgcatg
     3841 atgcctacct gaatgcagtg gtaggcactg cacttatcaa aaaatatccc aagcttgaat
     3901 ctgaatttgt ttacggagac tataaagtgt acgatgttag gaaaatgatc gcaaagtctg
     3961 agcaggaaat aggcaaggcc accgctaagt acttctttta cagcaatatt atgaattttt
     4021 tcaagaccga gattacactg gccaatggag agattcggaa gcgaccactt atcgaaacaa
     4081 acggagaaac aggagaaatc gtgtgggaca agggtaggga tttcgcgaca gtccggaagg
     4141 tcctgtccat gccgcaggtg aacatcgtta aaaagaccga agtacagacc ggaggcttct
     4201 ccaaggaaag tatcctcccg aaaaggaaca gcgacaagct gatcgcacgc aaaaaagatt
     4261 gggaccccaa gaaatacggc ggattcgatt ctcctacagt cgcttacagt gtactggttg
     4321 tggccaaagt ggagaaaggg aagtctaaaa aactcaaaag cgtcaaggaa ctgctgggca
     4381 tcacaatcat ggagcgatca agcttcgaaa aaaaccccat cgactttctc gaggcgaaag
     4441 gatataaaga ggtcaaaaaa gacctcatca ttaagcttcc caagtactct ctctttgagc
     4501 ttgaaaacgg ccggaaacga atgctcgcta gtgcgggcga gctgcagaaa ggtaacgagc
     4561 tggcactgcc ctctaaatac gttaatttct tgtatctggc cagccactat gaaaagctca
     4621 aagggtctcc cgaagataat gagcagaagc agctgttcgt ggaacaacac aaacactacc
     4681 ttgatgagat catcgagcaa ataagcgaat tctccaaaag agtgatcctc gccgacgcta
     4741 acctcgataa ggtgctttct gcttacaata agcacaggga taagcccatc agggagcagg
     4801 cagaaaacat tatccacttg tttactctga ccaacttggg cgcgcctgca gccttcaagt
     4861 acttcgacac caccatagac agaaagcggt acacctctac aaaggaggtc ctggacgcca
     4921 cactgattca tcagtcaatt acggggctct atgaaacaag aatcgacctc tctcagctcg
     4981 gtggagacag cagggctgac gggggaggat cagggggagg gtcccccaaa aagaaacgca
     5041 aagtgaccgg caagaaaacc agcggccagg ccggacaggc ctctcctaag aaaaaaagaa
     5101 aagtgggacg cgctggcggc ggaagcgaag aacttttgag caagaattat catcttgaga
     5161 acgaagtggc tcgtcttaag aaaggttctg gcagtggaga agaactgctt tcaaagaatt
     5221 accacctgga aaatgaggta gctagactga aaaaggggag cggaagtggg gaggagttgc
     5281 tgagcaaaaa ttatcatttg gagaacgaag tagcacgact aaagaaaggg tccggatcgg
     5341 gtgaggagtt actctcgaaa aattatcatc tcgaaaacga agtggctcgg ctaaaaaagg
     5401 gcagtggttc tggagaagag ctattatcta aaaactacca cctcgaaaat gaggtggcac
     5461 gcttaaaaaa gggaagtggc agtggtgaag agctactatc caagaattat catcttgaga
     5521 acgaggtagc gcgtttgaag aagggttccg gctcaggaga ggaactgctc tcgaagaact
     5581 atcatcttga aaatgaggtc gctcgattaa aaaagggatc gggcagtggt gaggaactac
     5641 tttcaaagaa ttaccacctc gaaaacgaag tagctcgatt aaagaaaggt tcagggtcgg
     5701 gtgaagaatt actgagtaaa aattatcatc tggaaaatga ggtagcgaga ctaaaaaagg
     5761 ggagtggttc tggcgaggaa ttgctatcga aaaattatca tcttgagaac gaagttgcta
     5821 ggctcaaaaa gggcggtggt tccggaggag gtagtccgaa gaagaaacgt aaggtcggat
     5881 gagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     5941 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     6001 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     6061 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     6121 agccaacgtc ggggcggcag gccctgccat agtgcaaaga tggataaagt tttaaacaga
     6181 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg
     6241 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     6301 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     6361 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     6421 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
     6481 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
     6541 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
     6601 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg
     6661 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
     6721 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
     6781 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
     6841 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
     6901 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
     6961 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
     7021 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
     7081 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
     7141 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
     7201 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
     7261 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
     7321 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
     7381 aagttttttt cttccatttc aggtgtcgtg acgtacggcc accatggtgt ctaagggcga
     7441 agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tggacaacca
     7501 tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag
     7561 aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctactagctt
     7621 cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact tcttcaagca
     7681 gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct
     7741 gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag
     7801 aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc
     7861 cttcaccgag acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg acatggccct
     7921 gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag accacatata gatccaagaa
     7981 acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac tggaaagaat
     8041 caaggaggcc aacaacgaga cctacgtcga gcagcacgag gtggcagtgg ccagatactg
     8101 cgacctccct agcaaactgg ggcacaagct taattgatgg aattaattcg ctgtctgcga
     8161 gggccagctg ttggggtgag tactccctct caaaagcggg catgacttct gcgctaagat
     8221 tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgcggtg atgcctttga
     8281 gggtggccgc gtccatctgg tcagaaaaga caatcttttt gttgtcaagc ttgaggtgtg
     8341 gcaggcttga gatctggcca tacacttgag tgacaatgac atccactttg cctttctctc
     8401 cacaggtgtc cactcccagg tccaactgca ggtcgagata actgatcata atcagccata
     8461 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     8521 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     8581 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     8641 gtggtttgtc caaactcatc aatgtatctt aatgcgtcga gtgcattcta gttgtggttt
     8701 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     8761 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     8821 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     8881 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     8941 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     9001 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     9061 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     9121 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     9181 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     9241 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     9301 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     9361 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     9421 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     9481 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     9541 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     9601 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     9661 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     9721 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     9781 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     9841 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     9901 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     9961 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
    10021 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
    10081 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
    10141 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
    10201 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
    10261 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
    10321 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
    10381 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
    10441 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
    10501 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
    10561 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
    10621 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
    10681 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
    10741 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
    10801 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
    10861 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
    10921 acctgacgtc
//
