LOCUS       sequence-345615-       10031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the dCas9-10xSunTag-tagBFP fusion protein for the 
            recruitment of scFv(GCN4)-fused effector domains. A shorter version 
            of the Addgene 60903 plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10031)
  AUTHORS   Hofacker D, Broche J, Laistner L, Adam S, Bashtrykov P, Jeltsch A
  TITLE     Engineering of Effector Domains for Targeted DNA Methylation with 
            Reduced Off-Target Effects.
  JOURNAL   Int J Mol Sci. 2020 Jan 13;21(2). pii: ijms21020502. doi: 
            10.3390/ijms21020502.
  PUBMED    31941101
REFERENCE   2  (bases 1 to 10031)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Int J Mol 
            Sci."; date: "2020-01-13"; volume: "21(2). pii: ijms21020502. doi"; 
            pages: " 10.3390/ijms21020502"
COMMENT     Sequence Label: dCas9-10xSunTag-P2A-tagBFP
FEATURES             Location/Qualifiers
     source          1..10031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             117..137
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      186..195
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(192..218,219..221,222..2708,2709..2711,2712..4295)
                     /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             192..4295
                     /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             4299..4325
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4305..4331
                     /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             4344..4364
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4371..4391
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4455..4511
                     /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             4527..4583
                     /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             4599..4655
                     /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             4671..4727
                     /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             4743..4799
                     /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             4815..4871
                     /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             4887..4943
                     /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             4959..5015
                     /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             5031..5087
                     /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             5103..5159
                     /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             5241..5261
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5271..5327
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5340..6038
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             6048..6068
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    6090..6678
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6143..6163)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6561..6572)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6777..7010
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(7035..7053)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7036..7053)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7343..7362)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(7576..7595)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7695..7717
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7755..7773)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7841..7945
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7946..8014,8015..8806)
                     /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(8164..8183)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8977..9565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9466..9485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9719..9736
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(9806..9826)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(9824..10031,1..90)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        join(9922..10031,1..101)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      join(9991..10031,1..95)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
       61 attccagaag tagtgaggag gcttttttgg aggcctaggc tacgcgcgcc accatgccca
      121 agaagaagcg caaggtggga cgcgtctgca ggatatcaag cttgcggtac cgcgggcccg
      181 ggatcgccac catggacaag aagtacagca tcggcctggc catcggcacc aactctgtgg
      241 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
      301 acaccgaccg gcacagcatc aagaagaacc tgatcggcgc cctgctgttc gacagcggag
      361 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
      421 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
      481 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
      541 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
      601 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
      661 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
      721 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
      781 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
      841 gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
      901 tgttcggcaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
      961 acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
     1021 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
     1081 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
     1141 tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag
     1201 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
     1261 acggctacgc cggctacatc gatggcggag ccagccagga agagttctac aagttcatca
     1321 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg
     1381 acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg
     1441 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
     1501 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
     1561 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
     1621 acttcgagga agtggtggac aagggcgcca gcgcccagag cttcatcgag cggatgacca
     1681 acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
     1741 acttcaccgt gtacaacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
     1801 ccgccttcct gagcggcgag cagaaaaaag ccatcgtgga cctgctgttc aagaccaacc
     1861 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
     1921 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
     1981 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
     2041 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
     2101 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
     2161 ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
     2221 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga
     2281 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
     2341 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
     2401 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
     2461 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
     2521 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
     2581 aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
     2641 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
     2701 atgtggacgc tatcgtgcct cagagctttc tgaaggacga ctccatcgat aacaaagtgc
     2761 tgactcggag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
     2821 agaagatgaa gaactactgg cgccagctgc tgaatgccaa gctgattacc cagaggaagt
     2881 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
     2941 tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact
     3001 cccggatgaa cactaagtac gacgagaacg acaaactgat ccgggaagtg aaagtgatca
     3061 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
     3121 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc
     3181 tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg
     3241 acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact
     3301 tcttctacag caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga
     3361 tccggaagcg gcctctgatc gagacaaacg gcgaaacagg cgagatcgtg tgggataagg
     3421 gccgggactt tgccaccgtg cggaaagtgc tgtctatgcc ccaagtgaat atcgtgaaaa
     3481 agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg
     3541 acaagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc
     3601 ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac
     3661 tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga
     3721 atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca
     3781 agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg
     3841 ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt
     3901 acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc
     3961 tgtttgtgga acagcacaaa cactacctgg acgagatcat cgagcagatc agcgagttct
     4021 ccaagagagt gatcctggcc gacgctaatc tggacaaggt gctgagcgcc tacaacaagc
     4081 acagagacaa gcctatcaga gagcaggccg agaatatcat ccacctgttt accctgacca
     4141 atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca
     4201 ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg
     4261 agacacggat cgacctgtct cagctgggag gcgacgccta tccctatgac gtgcccgatt
     4321 atgccagcct gggcagcggc tcccccaaga aaaaacgcaa ggtggaagat cctaagaaaa
     4381 agcggaaagt ggacggcatt ggtagtggga gcaacggcag cagcggatcc aacggtccga
     4441 ctgacgccgc ggaagaagaa cttttgagca agaattatca tcttgagaac gaagtggctc
     4501 gtcttaagaa aggttctggc agtggagaag aactgctttc aaagaattac cacctggaaa
     4561 atgaggtagc tagactgaaa aaggggagcg gaagtgggga ggagttgctg agcaaaaatt
     4621 atcatttgga gaacgaagta gcacgactaa agaaagggtc cggatcgggt gaggagttac
     4681 tctcgaaaaa ttatcatctc gaaaacgaag tggctcggct aaaaaagggc agtggttctg
     4741 gagaagagct attatctaaa aactaccacc tcgaaaatga ggtggcacgc ttaaaaaagg
     4801 gaagtggcag tggtgaagag ctactatcca agaattatca tcttgagaac gaggtagcgc
     4861 gtttgaagaa gggttccggc tcaggagagg aactgctctc gaagaactat catcttgaaa
     4921 atgaggtcgc tcgattaaaa aagggatcgg gcagtggtga ggaactactt tcaaagaatt
     4981 accacctcga aaacgaagta gctcgattaa agaaaggttc agggtcgggt gaagaattac
     5041 tgagtaaaaa ttatcatctg gaaaatgagg tagcgagact aaaaaagggg agtggttctg
     5101 gcgaggaatt gctatcgaaa aattatcatc ttgagaacga agttgctagg ctcaaaaagg
     5161 gctcaggctc aggcaccgcg gtaaacatag gtggtggaac cggtccgatg gatctacagc
     5221 ggccgcaagg tggaggtgga cccaagaaga agcgcaaggt gggaagcgga gctactaact
     5281 tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctgca ccaggaagta
     5341 tgagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtggaca
     5401 accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     5461 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacta
     5521 gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc gacttcttca
     5581 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
     5641 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
     5701 tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     5761 aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga aacgacatgg
     5821 ccctgaagct cgtgggcggg agccatctga tcgcaaacat caagaccaca tatagatcca
     5881 agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac agactggaaa
     5941 gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca gtggccagat
     6001 actgcgacct ccctagcaaa ctggggcaca agcttaatgg tggaggtcca aaaaagaaaa
     6061 ggaaggtcta acctgcaggg cttatcgata atcaacctct ggattacaaa atttgtgaaa
     6121 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     6181 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     6241 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     6301 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     6361 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     6421 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     6481 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     6541 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     6601 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     6661 tttgggccgc ctccccgcat ggtacctgca gggatcgatc actgaggtac ctttaagacc
     6721 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     6781 agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
     6841 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     6901 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     6961 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gcatctagaa
     7021 ttaattccgt gtattctata gtgtcaccta aatcgtatgt gtatgataca taaggttatg
     7081 tattaattgt agccgcgttc taacgacaat atgtacaagc ctaattgtgt agcatctggc
     7141 ttactgaagc agaccctatc atctctctcg taaactgccg tcagagtcgg tttggttgga
     7201 cgaaccttct gagtttctgg taacgccgtc ccgcacccgg aaatggtcag cgaaccaatc
     7261 agcagggtca tcgctagcca gatcctctac gccggacgca tcgtggccgg catcaccggc
     7321 gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct
     7381 cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc
     7441 gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac
     7501 ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga
     7561 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     7621 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     7681 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     7741 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     7801 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     7861 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     7921 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     7981 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     8041 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     8101 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     8161 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     8221 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     8281 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     8341 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     8401 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     8461 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     8521 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     8581 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     8641 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     8701 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     8761 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     8821 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     8881 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     8941 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     9001 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     9061 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     9121 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     9181 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     9241 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     9301 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     9361 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     9421 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     9481 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     9541 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     9601 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     9661 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     9721 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     9781 ttggccgatt cattaatgca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     9841 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     9901 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     9961 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
    10021 cattctccgc c
//
