LOCUS       sequence-365997-       10262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding SETD2-Cas9 fusion under CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10262)
  AUTHORS   Chen E, Lin-Shiao E, Trinidad M, Saffari Doost M, Colognori D, 
            Doudna JA
  TITLE     Decorating chromatin for enhanced genome editing using CRISPR-Cas9.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 Dec 6;119(49):e2204259119. doi: 
            10.1073/pnas.2204259119. Epub 2022 Dec 2.
  PUBMED    36459645
REFERENCE   2  (bases 1 to 10262)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2022-12-6"; volume: "119(49):e2204259119. 
            doi"; pages: " 10.1073/pnas.2204259119. Epub 2022 Dec 2"
COMMENT     Sequence Label: SETD2(SET)-Cas9
FEATURES             Location/Qualifiers
     source          1..10262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2379..6482
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2379..6482
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6486..6551
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6570..6590
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6597..6617
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6618..6674
                     /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             6675..7271
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7168..7188
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(7284..7303)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7349..7404
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7403..7422)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7765..7781)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7765..7781)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7778..7800)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7789..7805
                     /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(7813..7819,7820..7837,7838..7843))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7858..7879
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7938..8133
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7984..8119
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    8139..8273
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8176..8195)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8230..8249
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8341..8358)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8512..9100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8592..8611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9271..10062,10063..10131))
                     /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     9894..9913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10132..10236)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggnsgcg cgccrggggg gggggggggg ggsggggggg
      541 ggggsggggs grggsggrga ggkgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg grgggctcgg gggrggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcgccacca
     1741 tgattaagcg aatgcagtgt gagtgtacac ctctttctaa agatgaaaga gctcaaggtg
     1801 aaatagcatg tggggaagat tgtcttaatc gtcttctcat gattgaatgt tcttctcggt
     1861 gtccaaatgg ggattattgt tccaatagac ggtttcagag aaaacagcat gcagatgtgg
     1921 aagtcatact cacagaaaag aaaggctggg gcttgagagc tgccaaagac cttccttcga
     1981 acacctttgt cctagaatat tgtggagagg tactcgatca taaagagttt aaagctcgag
     2041 tgaaggagta tgcacgaaac aaaaacatcc attactattt catggccctg aagaatgatg
     2101 agataataga tgccactcaa aaaggaaatt gctctcgttt catgaatcac agctgtgaac
     2161 caaattgtga aacccaaaaa tggactgtga acggacaact gagggttggg ttttttacca
     2221 ccaaactggt tccttcaggc tcagagttaa cgtttgacta tcagttccag agatatggaa
     2281 aagaagccca gaaatgtttc tgcggatcag ccaattgccg gggttacctg ggaggtggta
     2341 gtggaggttc aggaggatcc ggggggagcg gagggagcat ggacaagaag tacagcatcg
     2401 gcctggacat cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc
     2461 ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga
     2521 tcggcgccct gctgttcgac agcggagaaa cagccgaggc cacccggctg aagagaaccg
     2581 ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca
     2641 acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg
     2701 aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct
     2761 accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca
     2821 aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact
     2881 tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc
     2941 tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg
     3001 ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc
     3061 agctgcccgg cgagaagaag aatggcctgt tcggcaacct gattgccctg agcctgggcc
     3121 tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca
     3181 aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg
     3241 acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag
     3301 tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc
     3361 accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca
     3421 aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacatcgat ggcggagcca
     3481 gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg
     3541 aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg
     3601 gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag
     3661 atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca
     3721 tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa
     3781 agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgccagcg
     3841 cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc
     3901 tgcccaagca cagcctgctg tacgagtact tcaccgtgta caacgagctg accaaagtga
     3961 aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaagcca
     4021 tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact
     4081 acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca
     4141 acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg
     4201 acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg
     4261 acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga
     4321 tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca
     4381 acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct
     4441 tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca
     4501 tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg
     4561 ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg
     4621 tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc
     4681 agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca
     4741 tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga
     4801 acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac
     4861 tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag agctttctga
     4921 aggacgactc catcgataac aaagtgctga ctcggagcga caagaaccgg ggcaagagcg
     4981 acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgc cagctgctga
     5041 atgccaagct gattacccag aggaagttcg acaatctgac caaggccgag agaggcggcc
     5101 tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca
     5161 caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac gagaacgaca
     5221 aactgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga
     5281 aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct
     5341 acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt
     5401 tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg
     5461 aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga
     5521 ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg
     5581 aaacaggcga gatcgtgtgg gataagggcc gggactttgc caccgtgcgg aaagtgctgt
     5641 ctatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag
     5701 agtctatcct gcccaagagg aacagcgaca agctgatcgc cagaaagaag gactgggacc
     5761 ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca
     5821 aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca
     5881 tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca
     5941 aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa
     6001 acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc
     6061 tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct
     6121 cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaaacac tacctggacg
     6181 agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg
     6241 acaaggtgct gagcgcctac aacaagcaca gagacaagcc tatcagagag caggccgaga
     6301 atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg
     6361 acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga
     6421 tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg
     6481 acgccgacta taaggaccac gacggagact acaaggatca tgatattgat tacaaagacg
     6541 atgacgataa gagcctgggc agcggctccc ccaagaaaaa acgcaaggtg gaagatccta
     6601 agaaaaagcg gaaagtggca acaaacttct ctctgctgaa acaagccgga gatgtcgaag
     6661 agaatcctgg accgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     6721 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     6781 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     6841 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     6901 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     6961 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     7021 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggagt ctcgcccgac caccagggca
     7081 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     7141 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     7201 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     7261 ccggtgcctg ataagctagc agatcttttt ccctctgcca aaaattatgg ggacatcatg
     7321 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     7381 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     7441 catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg
     7501 aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
     7561 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
     7621 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
     7681 ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg
     7741 cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     7801 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     7861 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     7921 gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     7981 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     8041 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     8101 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata
     8161 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     8221 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca
     8281 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     8341 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     8401 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     8461 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     8521 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     8581 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     8641 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     8701 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     8761 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     8821 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     8881 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     8941 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     9001 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     9061 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     9121 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     9181 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     9241 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     9301 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     9361 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     9421 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     9481 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     9541 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     9601 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     9661 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     9721 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     9781 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     9841 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     9901 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     9961 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
    10021 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
    10081 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
    10141 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
    10201 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
    10261 tg
//
