LOCUS       sequence-432326-       10827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPRcharm expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10827)
  AUTHORS   Neumann EN, Bertozzi TM, Wu E, Serack F, Harvey JW, Brauer PP, 
            Pirtle CP, Coffey A, Howard M, Kamath N, Lenz K, Guzman K, Raymond 
            MH, Khalil AS, Deverman BE, Minikel EV, Vallabh SM, Weissman JS
  TITLE     Brainwide silencing of prion protein by AAV-mediated delivery of an 
            engineered compact epigenetic editor.
  JOURNAL   Science. 2024 Jun 28;384(6703):ado7082. doi: 
            10.1126/science.ado7082. Epub 2024 Jun 28.
  PUBMED    38935715
REFERENCE   2  (bases 1 to 10827)
  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: "Science."; 
            date: "2024-06-28"; volume: "384(6703):ado7082. doi"; pages: " 
            10.1126/science.ado7082. Epub 2024 Jun 28"
COMMENT     Sequence Label: pENN226_CRISPRcharm
FEATURES             Location/Qualifiers
     source          1..10827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..658
                     /label=chicken beta-actin promoter
     intron          659..1667
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1591..1613
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1675..1694
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1760..1769
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2858..2878
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2879..2905,2906..2908,2909..5395,5396..5398,
                     5399..6982)
                     /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             2879..6982
                     /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             6983..7003
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7004..7060
                     /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"
     primer_bind     complement(7848..7867)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7913..7968
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7967..7986)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8327..8343)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8327..8343)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8340..8362)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8351..8367
                     /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(8375..8381,8382..8399,8400..8405))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8420..8441
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8500..8695
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8546..8681
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    8701..8835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8738..8757)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8792..8811
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8903..8920)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9074..9662)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9154..9173)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9833..10624,10625..10693))
                     /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     10456..10475
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10694..10798)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ggtcgaggtg agccccacgt tctgcttcac tctccccatc
      421 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
      481 atgggggcgg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      661 agtcgctgcg ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg
      721 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      781 ctgtaattag cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct
      841 taaagggctc cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt
      901 gtgtgtgcgt ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg
      961 gcgcggcgcg gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt
     1021 gccccgcggt gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
     1081 gggggtgagc agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct
     1141 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg
     1201 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     1261 cctcgggccg gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc
     1321 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     1381 ttcctttgtc ccaaatctgg cggagccgaa atctgggagg cgccgccgca ccccctctag
     1441 cgggcgcggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     1501 gcgtcgccgc gccgccgtcc ccttctccat ctccagcctc ggggctgccg cagggggacg
     1561 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1621 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1681 tgctggttat tgtgctgtct catcattttg gcaaagaatt ctgcagtcga cggtaccgcg
     1741 ggcccgggat ccaccggtcg ccaccatggc caggaccaaa caaactgcca gaaagagcac
     1801 cggggggaaa gcgccccgaa aacagctggc gaccaaagca gcccgcaaaa gcgcaccggg
     1861 aggtgcaagc agtggggcag gaagttcctc aggaggaagt gcggcgggat ccggaagcgg
     1921 tggcgcatca agtggcgccg caagtagcag tgccgggtcc gcagctggct ccgggtcctc
     1981 ccctgttgag atctacaaga ccgtgtccgc ctggaagcgg cagccaatca gagtgctgag
     2041 cctgttcggc aatattgaca aggaactgaa gtctctgggc ttcctggaaa gcggcagcca
     2101 cagcgagggc ggcacactga agtacgtgga agatgtgacc aacgtggtgc ggagagatgt
     2161 ggaaaagtgg ggacctttcg acctgctgta cggcagcacc cagcctctgg gcggaagctg
     2221 cgacaggtgt cctggctggt acatgttcca atttcataga atcctgcagt acgccagacc
     2281 tagacaggag tctcagccct tcttctggat cttcatggat aacctgctgc tcacagagga
     2341 cgaccagatg accaccgtgc ggttcctgca aacagaggcc gtcaccctgc gcgacgtgag
     2401 atctagagtg ctgcagaacg ccgtccgggt gtggagcaac atccccggtc ttaaaagcaa
     2461 gcacgaggct ctgtccccaa aagaggaaga gagcctgcag ggccacgtgc ggaccagatc
     2521 taagctggcc gctcctaaag tggaccccct ggtgaagaac tgcctgctgc ctctgagaga
     2581 gtactttaag tatttcagcc agagcagcct gcccctgggc ggcccttcta gcggtgctcc
     2641 acctccaagc ggcggctcgc cagccggcag ccctaccagc accgaggagg gcacaagcga
     2701 gagcgcaaca cctgagagcg gccccggcac aagcaccgag ccttctgaag gaagcgcccc
     2761 tgggagccct gccggcagcc ccaccagcac agaagagggc acctctaccg agcccagcga
     2821 gggtagcgcc cctggcacaa gcacagaacc ctctgagcct aagaaaaagc ggaaggtgat
     2881 ggacaagaag tacagcatcg gcctggccat cggcaccaac tctgtgggct gggccgtgat
     2941 caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca
     3001 cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc
     3061 cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta
     3121 tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct tccacagact
     3181 ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa
     3241 catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa
     3301 actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat
     3361 gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt
     3421 ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat
     3481 caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg
     3541 gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct
     3601 gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga
     3661 tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca
     3721 gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct
     3781 gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat
     3841 gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca
     3901 gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg
     3961 ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga
     4021 aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa
     4081 gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc
     4141 cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga
     4201 gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag
     4261 attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt
     4321 ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa
     4381 cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta
     4441 taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag
     4501 cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt
     4561 gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc
     4621 cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat
     4681 caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct
     4741 gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca
     4801 cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag
     4861 gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga
     4921 tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag
     4981 cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca
     5041 cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt
     5101 gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat
     5161 cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat
     5221 gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt
     5281 ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga
     5341 tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggacgctat
     5401 cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga
     5461 caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa
     5521 ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac
     5581 caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct
     5641 ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac
     5701 taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa
     5761 gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta
     5821 ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta
     5881 ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat
     5941 gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa
     6001 catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc
     6061 tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc
     6121 caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca
     6181 gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc
     6241 cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta
     6301 ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa
     6361 agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt
     6421 tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta
     6481 ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca
     6541 gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca
     6601 ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca
     6661 gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat
     6721 cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc
     6781 catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc
     6841 tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga
     6901 ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga
     6961 cctgtctcag ctgggaggtg accccaagaa gaagcggaag gtggccacaa atttcagcct
     7021 gctgaagcag gccggcgacg tggaagagaa tcctggcccc agcgagctga tcaaagagaa
     7081 catgcacatg aagctgtaca tggaaggcac cgtggacaac catcacttca agtgcaccag
     7141 cgagggcgag ggcaagccct acgagggcac acagaccatg agaatcaagg tggtggaagg
     7201 aggcccactg cccttcgcct tcgacatcct ggccaccagc tttctgtacg gctcaaagac
     7261 cttcatcaac cacacccagg gcatccctga tttcttcaag cagagcttcc ctgagggatt
     7321 tacctgggag agagtgacaa cttatgaaga tggcggagtg ctgaccgcca cacaagatac
     7381 cagcctgcag gacggctgtc tgatctacaa cgtgaagatc agaggcgtga acttcaccag
     7441 caacggccct gtgatgcaga aaaaaaccct gggctgggag gcctttacag agacactgta
     7501 ccccgctgat ggaggcctgg aaggtagaaa cgacatggcc ctgaaactgg tcggcggcag
     7561 ccacctgatt gccaatatca agaccacata ccggagcaag aaaccagcta aaaacctgaa
     7621 gatgcctggc gtttactacg tggattacag actggagcgg atcaaggaag ccaacaacga
     7681 gacatatgtg gaacagcacg aggtggccgt agctagatac tgcgacctgc cttctaagct
     7741 tggccacaag ctgaactgat aagcggccgc actcctcagg tgcaggctgc ctatcagaag
     7801 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
     7861 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     7921 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     7981 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     8041 tatgccatat gctggctgcc atgaacaaag gtggctataa agaggtcatc agtatatgaa
     8101 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     8161 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     8221 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     8281 cttatgaaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
     8341 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     8401 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     8461 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac
     8521 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     8581 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     8641 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     8701 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     8761 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     8821 tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     8881 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     8941 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     9001 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     9061 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     9121 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     9181 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     9241 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     9301 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     9361 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     9421 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     9481 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     9541 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     9601 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     9661 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     9721 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     9781 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     9841 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     9901 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     9961 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
    10021 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
    10081 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
    10141 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
    10201 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
    10261 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    10321 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    10381 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    10441 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    10501 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    10561 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    10621 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    10681 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    10741 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    10801 catttccccg aaaagtgcca cctggtc
//
