LOCUS       sequence-267134-        9420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of fusion protein M3-M14-dCas9 in Mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9420)
  AUTHORS   Liu XM, Zhou J, Mao Y, Ji Q, Qian SB
  TITLE     Programmable RNA N(6)-methyladenosine editing by CRISPR-Cas9 
            conjugates.
  JOURNAL   Nat Chem Biol. 2019 Sep;15(9):865-871. doi: 
            10.1038/s41589-019-0327-1. Epub 2019 Aug 5.
  PUBMED    31383972
REFERENCE   2  (bases 1 to 9420)
  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: 
            "10.1038/s41589-019-0327-1"; journalName: "Nat Chem Biol"; date: 
            "2019-09"; volume: "15"; issue: "9"; pages: "865-871"
COMMENT     Sequence Label: M3-M14-dCas9
FEATURES             Location/Qualifiers
     source          1..9420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(569..3967,3968..3970,3971..4567,4568,4569..4571,
                     4572..4573,4574..4576,4577..4669)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(569..592,593..595,596..3082,3083..3085,3086..4669)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4682..4930
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             4943..4963
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4972..4989
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5012..5029)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5018..5242
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5313..5329)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5313..5329)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5326..5348)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5337..5353
                     /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(5361..5367,5368..5385,5386..5391))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5406..5427
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5544..5561)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5715..6303)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5795..5814)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6474..7265,7266..7334))
                     /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     7097..7116
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7335..7439)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7518..7537)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7709..8088
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8089..8292
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8243..8263
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8334..8353
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8334..8352
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tggttctggg gaggggttgg accttggaag agtgtgttta cgaaaatggg gttacagaag
       61 atgtgaagat atttgttgga ttaaaaccaa taaaaacaat cctgggaaga ctaagacttt
      121 agatccaaag gctgtctttc agagaacaaa ggaacactgc ctcatgggga tcaaaggaac
      181 tgtgaagcgt agcacagacg gggacttcat tcatgctaat gttgacattg acttaattat
      241 cacagaagaa cctgaaattg gcaatataga aaaacctgta gaaatttttc atataattga
      301 gcatttttgt cttggtagaa gacgccttca tctatttgga agagatagta caattcgacc
      361 aggctggctc acagttggac caacgcttac aaatagcaac tacaatgcag aaacatatgc
      421 atcctatttc agtgctccta attcctactt gactggttgt acagaagaaa ttgagagact
      481 tcgaccaaaa tcgcctcctc ccaaatctaa atctgaccga agcggcagcg agactcccgg
      541 gacctcagag tccgccacac ccgaaagtga taaaaagtat tctattggtt tagccatcgg
      601 cactaattcc gttggatggg ctgtcataac cgatgaatac aaagtacctt caaagaaatt
      661 taaggtgttg gggaacacag accgtcattc gattaaaaag aatcttatcg gtgccctcct
      721 attcgatagt ggcgaaacgg cagaggcgac tcgcctgaaa cgaaccgctc ggagaaggta
      781 tacacgtcgc aagaaccgaa tatgttactt acaagaaatt tttagcaatg agatggccaa
      841 agttgacgat tctttctttc accgtttgga agagtccttc cttgtcgaag aggacaagaa
      901 acatgaacgg caccccatct ttggaaacat agtagatgag gtggcatatc atgaaaagta
      961 cccaacgatt tatcacctca gaaaaaagct agttgactca actgataaag cggacctgag
     1021 gttaatctac ttggctcttg cccatatgat aaagttccgt gggcactttc tcattgaggg
     1081 tgatctaaat ccggacaact cggatgtcga caaactgttc atccagttag tacaaaccta
     1141 taatcagttg tttgaagaga accctataaa tgcaagtggc gtggatgcga aggctattct
     1201 tagcgcccgc ctctctaaat cccgacggct agaaaacctg atcgcacaat tacccggaga
     1261 gaagaaaaat gggttgttcg gtaaccttat agcgctctca ctaggcctga caccaaattt
     1321 taagtcgaac ttcgacttag ctgaagatgc caaattgcag cttagtaagg acacgtacga
     1381 tgacgatctc gacaatctac tggcacaaat tggagatcag tatgcggact tatttttggc
     1441 tgccaaaaac cttagcgatg caatcctcct atctgacata ctgagagtta atactgagat
     1501 taccaaggcg ccgttatccg cttcaatgat caaaaggtac gatgaacatc accaagactt
     1561 gacacttctc aaggccctag tccgtcagca actgcctgag aaatataagg aaatattctt
     1621 tgatcagtcg aaaaacgggt acgcaggtta tattgacggc ggagcgagtc aagaggaatt
     1681 ctacaagttt atcaaaccca tattagagaa gatggatggg acggaagagt tgcttgtaaa
     1741 actcaatcgc gaagatctac tgcgaaagca gcggactttc gacaacggta gcattccaca
     1801 tcaaatccac ttaggcgaat tgcatgctat acttagaagg caggaggatt tttatccgtt
     1861 cctcaaagac aatcgtgaaa agattgagaa aatcctaacc tttcgcatac cttactatgt
     1921 gggacccctg gcccgaggga actctcggtt cgcatggatg acaagaaagt ccgaagaaac
     1981 gattactcca tggaattttg aggaagttgt cgataaaggt gcgtcagctc aatcgttcat
     2041 cgagaggatg accaactttg acaagaattt accgaacgaa aaagtattgc ctaagcacag
     2101 tttactttac gagtatttca cagtgtacaa tgaactcacg aaagttaagt atgtcactga
     2161 gggcatgcgt aaacccgcct ttctaagcgg agaacagaag aaagcaatag tagatctgtt
     2221 attcaagacc aaccgcaaag tgacagttaa gcaattgaaa gaggactact ttaagaaaat
     2281 tgaatgcttc gattctgtcg agatctccgg ggtagaagat cgatttaatg cgtcacttgg
     2341 tacgtatcat gacctcctaa agataattaa agataaggac ttcctggata acgaagagaa
     2401 tgaagatatc ttagaagata tagtgttgac tcttaccctc tttgaagatc gggaaatgat
     2461 tgaggaaaga ctaaaaacat acgctcacct gttcgacgat aaggttatga aacagttaaa
     2521 gaggcgtcgc tatacgggct ggggacgatt gtcgcggaaa cttatcaacg ggataagaga
     2581 caagcaaagt ggtaaaacta ttctcgattt tctaaagagc gacggcttcg ccaataggaa
     2641 ctttatgcag ctgatccatg atgactcttt aaccttcaaa gaggatatac aaaaggcaca
     2701 ggtttccgga caaggggact cattgcacga acatattgcg aatcttgctg gttcgccagc
     2761 catcaaaaag ggcatactcc agacagtcaa agtagtggat gagctagtta aggtcatggg
     2821 acgtcacaaa ccggaaaaca ttgtaatcga gatggcacgc gaaaatcaaa cgactcagaa
     2881 ggggcaaaaa aacagtcgag agcggatgaa gagaatagaa gagggtatta aagaactggg
     2941 cagccagatc ttaaaggagc atcctgtgga aaatacccaa ttgcagaacg agaaacttta
     3001 cctctattac ctacaaaatg gaagggacat gtatgttgat caggaactgg acataaaccg
     3061 tttatctgat tacgacgtcg atgccattgt accccaatcc tttttgaagg acgattcaat
     3121 cgacaataaa gtgcttacac gctcggataa gaaccgaggg aaaagtgaca atgttccaag
     3181 cgaggaagtc gtaaagaaaa tgaagaacta ttggcggcag ctcctaaatg cgaaactgat
     3241 aacgcaaaga aagttcgata acttaactaa agctgagagg ggtggcttgt ctgaacttga
     3301 caaggccgga tttattaaac gtcagctcgt ggaaacccgc caaatcacaa agcatgttgc
     3361 acagatacta gattcccgaa tgaatacgaa atacgacgag aacgataagc tgattcggga
     3421 agtcaaagta atcactttaa agtcaaaatt ggtgtcggac ttcagaaagg attttcaatt
     3481 ctataaagtt agggagataa ataactacca ccatgcgcac gacgcttatc ttaatgccgt
     3541 cgtagggacc gcactcatta agaaataccc gaagctagaa agtgagtttg tgtatggtga
     3601 ttacaaagtt tatgacgtcc gtaagatgat cgcgaaaagc gaacaggaga taggcaaggc
     3661 tacagccaaa tacttctttt attctaacat tatgaatttc tttaagacgg aaatcactct
     3721 ggcaaacgga gagatacgca aacgaccttt aattgaaacc aatggggaga caggtgaaat
     3781 cgtatgggat aagggccggg acttcgcgac ggtgagaaaa gttttgtcca tgccccaagt
     3841 caacatagta aagaaaactg aggtgcagac cggagggttt tcaaaggaat cgattcttcc
     3901 aaaaaggaat agtgataagc tcatcgctcg taaaaaggac tgggacccga aaaagtacgg
     3961 tggcttcgat agccctacag ttgcctattc tgtcctagta gtggcaaaag ttgagaaggg
     4021 aaaatccaag aaactgaagt cagtcaaaga attattgggg ataacgatta tggagcgctc
     4081 gtcttttgaa aagaacccca tcgacttcct tgaggcgaaa ggttacaagg aagtaaaaaa
     4141 ggatctcata attaaactac caaagtatag tctgtttgag ttagaaaatg gccgaaaacg
     4201 gatgttggct agcgccggag agcttcaaaa ggggaacgaa ctcgcactac cgtctaaata
     4261 cgtgaatttc ctgtatttag cgtcccatta cgagaagttg aaaggttcac ctgaagataa
     4321 cgaacagaag caactttttg ttgagcagca caaacattat ctcgacgaaa tcatagagca
     4381 aatttcggaa ttcagtaaga gagtcatcct agctgatgcc aatctggaca aagtattaag
     4441 cgcatacaac aagcacaggg ataaacccat acgtgagcag gcggaaaata ttatccattt
     4501 gtttactctt accaacctcg gcgctccagc cgcattcaag tattttgaca caacgataga
     4561 tcgcaaacga tacacttcta ccaaggaggt gctagacgcg acactgattc accaatccat
     4621 cacgggatta tatgaaactc ggatagattt gtcacagctt gggggtgact ctggtggttc
     4681 tactaatctg tcagatatta ttgaaaagga gaccggtaag caactggtta tccaggaatc
     4741 catcctcatg ctcccagagg aggtggaaga agtcattggg aacaagccgg aaagcgatat
     4801 actcgtgcac accgcctacg acgagagcac cgacgagaat gtcatgcttc tgactagcga
     4861 cgcccctgaa tacaagcctt gggctctggt catacaggat agcaacggtg agaacaagat
     4921 taagatgctc tctggtggtt ctcccaagaa gaagaggaaa gtctaaccgg tcatcatcac
     4981 catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     5041 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     5101 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     5161 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     5221 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctcgataccg
     5281 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5341 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctagggt
     5401 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5461 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5521 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5581 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5641 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5701 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5761 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5821 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5881 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5941 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6001 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6061 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6121 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6181 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6241 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6301 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6361 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6421 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6481 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6541 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6601 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6661 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6721 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6781 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6841 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6901 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6961 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7021 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7081 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7141 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7201 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7261 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7321 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7381 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7441 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcgatc tcccgatccc
     7501 ctagggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     7561 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     7621 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     7681 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     7741 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     7801 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     7861 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     7921 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     7981 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     8041 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     8101 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     8161 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     8221 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     8281 ataagcagag ctggtttagt gaaccgtcag atccgctaga gatccgcggc cgctaatacg
     8341 actcactata gggagagccg ccaccatgtt cccacctcag tggatctgtt gtgatatccg
     8401 ctacctggac gtcagtatct tgggcaagtt tgcagttgtg atggctgacc caccctggga
     8461 tattcacatg gaactgccct atgggaccct gacagatgat gagatgcgca ggctcaacat
     8521 acccgtacta caggatgatg gctttctctt cctctgggtc acaggcaggg ccatggagtt
     8581 ggggagagaa tgtctaaacc tctgggggta tgaacgggta gatgaaatta tttgggtgaa
     8641 gacaaatcaa ctgcaacgca tcattcggac aggccgtaca ggtcactggt tgaaccatgg
     8701 gaaggaacac tgcttggttg gtgtcaaagg aaatccccaa ggcttcaacc agggtctgga
     8761 ttgtgatgtg atcgtagctg aggttcgttc caccagtcat aaaccagatg aaatctatgg
     8821 catgattgaa agactatctc ctggcactcg caagattgag ttatttggac gaccacacaa
     8881 tgtgcaaccc aactggatca cccttggaaa ccaactggat gggatccacc tactagaccc
     8941 agatgtggtt gcacggttca agcaaaggta cccagatggt atcatctcta aacctaagaa
     9001 tttaggcgga tcaggtggct ccggagggtc tggtggatcg ggaacacaga gcttaaatcc
     9061 ccataatgat tactgccaac attttgtaga cactggacat agacctcaga atttcatcag
     9121 ggatgtaggt ttagctgaca gatttgaaga atatcctaaa ctgagggagc tcatcaggct
     9181 aaaggatgag ttaatagcta aatctaacac tcctcccatg tacttacaag ccgatataga
     9241 agcctttgac atcagagaac taacacccaa atttgatgtg attcttctgg aacccccttt
     9301 agaagaatat tacagagaaa ctggcatcac tgctaatgaa aaatgctgga cttgggatga
     9361 tattatgaag ttagaaattg atgagattgc agcacctcga tcatttattt ttctctggtg
//
