LOCUS       sequence-366024-       10463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding SETMAR-Cas9 fusion under CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10463)
  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 10463)
  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: SETMAR-Cas9
FEATURES             Location/Qualifiers
     source          1..10463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        13..392
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          671..1687
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1611..1633
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1695..1714
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2588..6691
                     /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             2588..6691
                     /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             6695..6760
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6779..6799
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6806..6826
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6827..6883
                     /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             6884..7480
                     /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     7377..7397
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(7493..7512)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7558..7613
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7612..7631)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7974..7990)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7974..7990)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7987..8009)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7998..8014
                     /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(8022..8028,8029..8046,8047..8052))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8067..8088
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8147..8342
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8193..8328
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    8348..8482
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8385..8404)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8439..8458
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8550..8567)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8721..9309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8801..8820)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9480..10271,10272..10340))
                     /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     10103..10122
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10341..10445)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccacctggg tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
       61 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      121 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      181 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      241 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      301 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      361 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
      421 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
      481 tttgtgcagc gatgggggcg gggggggggg gggnsgcgcg ccrggsgggg sggggsgggg
      541 sgrggggsgg ggsggggsgr ggcggrgagg tgcggcggca gccaatcaga gcggcgcgct
      601 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
      661 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
      721 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
      781 cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg
      841 gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg
      901 gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg
      961 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag
     1021 cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt
     1081 gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc
     1141 ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg
     1201 tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc
     1261 gggcggggcg gggccgcctc gggccggggr gggctcgggg gaggggcgcg gcggcccccg
     1321 gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg
     1381 cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg
     1441 ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat
     1501 gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg
     1561 gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt
     1621 ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt
     1681 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt
     1741 cgccaccatg ttcgcggagt ttaaggagaa gcctgaggcc ccgactgagc agctggatgt
     1801 cgcgtgcggc caggaaaact tgccggtggg cgcgtggccc ccgggggccg cgccggcgcc
     1861 cttccagtac actcctgatc atgtagttgg acctggagca gacattgatc ccactcaaat
     1921 aacctttccc ggatgcattt gtgtcaaaac tccctgcctc cctggcactt gctcctgtct
     1981 ccgccatgga gagaactatg atgataactc atgccttaga gatataggat ctggaggaaa
     2041 gtatgcagag cctgtttttg aatgcaatgt cctgtgccga tgcagtgacc actgcagaaa
     2101 cagagtggtc cagaaaggtc tacagttcca cttccaagtg ttcaagacgc ataaaaaagg
     2161 ctggggactt cgtaccttgg aatttatacc gaaaggaagg tttgtctgtg aatatgctgg
     2221 tgaggtttta ggattctctg aagttcagag aagaattcac ttacaaacaa aatccgactc
     2281 caattacatt atagccatca gggaacatgt ttataatggg caggtaatgg aaacatttgt
     2341 tgaccctact tatataggaa atattggaag attccttaat cattcttgtg agccaaacct
     2401 tttgatgatt cctgtccgaa ttgactcaat ggtacctaag ttggcacttt ttgcagccaa
     2461 agatattgtg ccagaagaag aactctctta tgattattca ggaagatatc ttaatctaac
     2521 agtcagtgaa gacaaagaag gcggtggtag tggaggttca ggaggatccg gggggagcgg
     2581 agggagcatg gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg
     2641 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
     2701 cgaccggcac agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggagaaac
     2761 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
     2821 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
     2881 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
     2941 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
     3001 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
     3061 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
     3121 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
     3181 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
     3241 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     3301 cggcaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     3361 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     3421 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     3481 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     3541 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     3601 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     3661 ctacgccggc tacatcgatg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     3721 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     3781 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     3841 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     3901 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     3961 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     4021 cgaggaagtg gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt
     4081 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     4141 caccgtgtac aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     4201 cttcctgagc ggcgagcaga aaaaagccat cgtggacctg ctgttcaaga ccaaccggaa
     4261 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     4321 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     4381 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     4441 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     4501 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     4561 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     4621 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     4681 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     4741 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     4801 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     4861 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
     4921 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
     4981 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     5041 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
     5101 ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgataaca aagtgctgac
     5161 tcggagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
     5221 gatgaagaac tactggcgcc agctgctgaa tgccaagctg attacccaga ggaagttcga
     5281 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
     5341 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
     5401 gatgaacact aagtacgacg agaacgacaa actgatccgg gaagtgaaag tgatcaccct
     5461 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
     5521 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
     5581 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
     5641 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     5701 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     5761 gaagcggcct ctgatcgaga caaacggcga aacaggcgag atcgtgtggg ataagggccg
     5821 ggactttgcc accgtgcgga aagtgctgtc tatgccccaa gtgaatatcg tgaaaaagac
     5881 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgacaa
     5941 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
     6001 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     6061 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     6121 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     6181 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
     6241 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     6301 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
     6361 tgtggaacag cacaaacact acctggacga gatcatcgag cagatcagcg agttctccaa
     6421 gagagtgatc ctggccgacg ctaatctgga caaggtgctg agcgcctaca acaagcacag
     6481 agacaagcct atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
     6541 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
     6601 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
     6661 acggatcgac ctgtctcagc tgggaggcga cgccgactat aaggaccacg acggagacta
     6721 caaggatcat gatattgatt acaaagacga tgacgataag agcctgggca gcggctcccc
     6781 caagaaaaaa cgcaaggtgg aagatcctaa gaaaaagcgg aaagtggcaa caaacttctc
     6841 tctgctgaaa caagccggag atgtcgaaga gaatcctgga ccgaccgagt acaagcccac
     6901 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     6961 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     7021 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     7081 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     7141 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     7201 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     7261 cgtcggagtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     7321 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     7381 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     7441 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga taagctagca gatctttttc
     7501 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     7561 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     7621 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     7681 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     7741 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     7801 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     7861 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     7921 ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag
     7981 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     8041 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     8101 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt
     8161 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     8221 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     8281 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     8341 aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     8401 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     8461 tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc
     8521 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     8581 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     8641 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     8701 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     8761 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     8821 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     8881 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     8941 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     9001 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     9061 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     9121 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     9181 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     9241 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     9301 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     9361 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     9421 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     9481 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     9541 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     9601 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     9661 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     9721 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     9781 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     9841 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     9901 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     9961 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
    10021 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
    10081 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
    10141 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
    10201 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
    10261 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
    10321 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
    10381 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
    10441 ccgcgcacat ttccccgaaa agt
//
