LOCUS       sequence-365996-       10780 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Plasmid encoding PRDM9dC-Cas9 fusion under CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10780)
  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 10780)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: PRDM9dC(KRAB-SSXRD-PR/SET-dC)-Cas9
FEATURES             Location/Qualifiers
     source          1..10780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2897..7000
                     /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             2897..7000
                     /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             7004..7069
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7088..7108
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7115..7135
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7136..7192
                     /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             7193..7789
                     /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     7686..7706
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(7802..7821)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7867..7922
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7921..7940)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8283..8299)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8283..8299)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8296..8318)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8307..8323
                     /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(8331..8337,8338..8355,8356..8361))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8376..8397
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8456..8651
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8502..8637
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    8657..8791
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8694..8713)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8748..8767
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8859..8876)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9030..9618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9110..9129)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9789..10580,10581..10649))
                     /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     10412..10431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10650..10754)
                     /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 gggggcgcgc gccrggsggg gggggggggg gggggggggg
      541 gggsggggsg rggsggrgrg gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg rgggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgccaccat
     1741 gagccctgaa aagtcccaag aggagagccc agaagaagac acagagagaa cagagcggaa
     1801 gcccatggtc aaagatgcct tcaaagacat ttccatatac ttcaccaagg aagaatgggc
     1861 agagatggga gactgggaga aaactcgcta taggaatgtg aaaaggaact ataatgcact
     1921 gattactata ggtctcagag ccactcgacc agctttcatg tgtcaccgaa ggcaggccat
     1981 caaactccag gtggatgaca cagaagattc tgatgaagaa tggaccccta ggcagcaagt
     2041 caaacctcct tggatggcct taagagtgga acagcgtaaa caccagaagg gaatgcccaa
     2101 ggcgtcattc agtaatgaat ctagtttgaa agaattgtca agaacagcaa atttactgaa
     2161 tgcaagtggc tcagagcagg ctcagaaacc agtgtcccct tctggagaag caagtacctc
     2221 tggacagcac tcaagactaa aactggaact caggaagaag gagactgaaa gaaagatgta
     2281 tagcctgcga gaaagaaagg gtcatgcata caaagaggtc agcgagccgc aggatgatga
     2341 ttacctctat tgtgagatgt gtcagaactt cttcattgac agctgtgctg cccatgggcc
     2401 ccctacattt gtaaaggaca gtgcagtgga caaggggcac cccaaccgtt cagccctcag
     2461 tctgccccca gggctgagaa ttgggccatc aggcatccct caggctgggc ttggagtatg
     2521 gaatgaggca tctgatctgc cgctgggtct gcactttggc ccttatgagg gccgaattac
     2581 agaagacgaa gaggcagcca acaatggata ctcctggctg atcaccaagg ggagaaactg
     2641 ctatgagtat gtggatggaa aagataaatc ctgggccaac tggatgaggt atgtgaactg
     2701 tgcccgggat gatgaagagc agaacctggt ggccttccag taccacaggc agatcttcta
     2761 tagaacctgc cgagtcatta ggccaggctg tgaactgctg gtctggtatg gggatgaata
     2821 cggccaggaa ctgggcatca agtggggcag cggtggtagt ggaggttcag gaggatccgg
     2881 ggggagcgga gggagcatgg acaagaagta cagcatcggc ctggacatcg gcaccaactc
     2941 tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct
     3001 gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggcgccctgc tgttcgacag
     3061 cggagaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg
     3121 gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga
     3181 cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg
     3241 gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat
     3301 ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta
     3361 tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa
     3421 ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct
     3481 gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag
     3541 actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa
     3601 tggcctgttc ggcaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa
     3661 cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct
     3721 ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa
     3781 cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc
     3841 ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct
     3901 gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag
     3961 caagaacggc tacgccggct acatcgatgg cggagccagc caggaagagt tctacaagtt
     4021 catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag
     4081 agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca
     4141 cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga
     4201 caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct
     4261 ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc
     4321 ctggaacttc gaggaagtgg tggacaaggg cgccagcgcc cagagcttca tcgagcggat
     4381 gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta
     4441 cgagtacttc accgtgtaca acgagctgac caaagtgaaa tacgtgaccg agggaatgag
     4501 aaagcccgcc ttcctgagcg gcgagcagaa aaaagccatc gtggacctgc tgttcaagac
     4561 caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt
     4621 cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca
     4681 cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat
     4741 tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg
     4801 gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag
     4861 atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc
     4921 cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca
     4981 gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg
     5041 ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa
     5101 gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa
     5161 gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa
     5221 gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat
     5281 cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta
     5341 cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga
     5401 ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgataacaa
     5461 agtgctgact cggagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt
     5521 cgtgaagaag atgaagaact actggcgcca gctgctgaat gccaagctga ttacccagag
     5581 gaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg
     5641 cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct
     5701 ggactcccgg atgaacacta agtacgacga gaacgacaaa ctgatccggg aagtgaaagt
     5761 gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt
     5821 gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac
     5881 cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt
     5941 gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa
     6001 gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg
     6061 cgagatccgg aagcggcctc tgatcgagac aaacggcgaa acaggcgaga tcgtgtggga
     6121 taagggccgg gactttgcca ccgtgcggaa agtgctgtct atgccccaag tgaatatcgt
     6181 gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa
     6241 cagcgacaag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga
     6301 cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa
     6361 gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga
     6421 gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat
     6481 catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc
     6541 ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt
     6601 cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa
     6661 acagctgttt gtggaacagc acaaacacta cctggacgag atcatcgagc agatcagcga
     6721 gttctccaag agagtgatcc tggccgacgc taatctggac aaggtgctga gcgcctacaa
     6781 caagcacaga gacaagccta tcagagagca ggccgagaat atcatccacc tgtttaccct
     6841 gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag
     6901 gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct
     6961 gtacgagaca cggatcgacc tgtctcagct gggaggcgac gccgactata aggaccacga
     7021 cggagactac aaggatcatg atattgatta caaagacgat gacgataaga gcctgggcag
     7081 cggctccccc aagaaaaaac gcaaggtgga agatcctaag aaaaagcgga aagtggcaac
     7141 aaacttctct ctgctgaaac aagccggaga tgtcgaagag aatcctggac cgaccgagta
     7201 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     7261 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     7321 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     7381 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     7441 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     7501 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     7561 cctggccacc gtcggagtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     7621 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     7681 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     7741 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgat aagctagcag
     7801 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     7861 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     7921 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     7981 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
     8041 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     8101 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
     8161 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     8221 tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca
     8281 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     8341 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     8401 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc
     8461 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     8521 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     8581 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     8641 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     8701 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     8761 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg
     8821 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     8881 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     8941 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     9001 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     9061 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     9121 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     9181 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     9241 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     9301 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     9361 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     9421 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     9481 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     9541 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     9601 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     9661 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     9721 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     9781 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     9841 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     9901 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     9961 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
    10021 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
    10081 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
    10141 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
    10201 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
    10261 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
    10321 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
    10381 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
    10441 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
    10501 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
    10561 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
    10621 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
    10681 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
    10741 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
