LOCUS       sequence-252320-       15158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9-hUtx-IRES-Hyg in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15158)
  AUTHORS   Ochiai H, Hayashi T, Umeda M, Yoshimura M, Harada A, Shimizu Y, 
            Nakano K, Saitoh N, Liu Z, Yamamoto T, Okamura T, Ohkawa Y, Kimura 
            H, Nikaido I
  TITLE     Genome-wide kinetic properties of transcriptional bursting in mouse 
            embryonic stem cells.
  JOURNAL   Sci Adv. 2020 Jun 17;6(25):eaaz6699. doi: 10.1126/sciadv.aaz6699. 
            eCollection 2020 Jun.
  PUBMED    32596448
REFERENCE   2  (bases 1 to 15158)
  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: "Sci Adv."; 
            date: "2020-06-17"; volume: "6(25):eaaz6699. doi"; pages: " 
            10.1126/sciadv.aaz6699. eCollection 2020 Jun"
COMMENT     Sequence Label: pLR5-CBh-dCas9-hUtx-IRES-Hyg
FEATURES             Location/Qualifiers
     source          1..15158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          113..340
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             361..387
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             391..411
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(421..444,445..447,448..3003,3004..3006,3007..4521)
                     /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 N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(421..444,445..447,448..2934,2935..2937,2938..4521)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4525..4545
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          8965..9194
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     9177..9196
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    9267..9840
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(9434..9451)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     9661..9680
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             9876..10901
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             9876..10901
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    11029..11150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11066..11085)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   11408..11470
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(11521..11541)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11521..11539)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11560..11576)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11560..11576)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11573..11595)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11584..11600
                     /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(11608..11614,11615..11632,11633..11638))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11653..11674
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11791..11808)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11962..12550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12042..12061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12721..13512,13513..13581))
                     /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     13344..13363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13582..13686)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(13712..14167)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(13849..13870)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     14061..14080
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14294..14316
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     14308..14325
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     14309..14325
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14335..14354
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        14335..14353
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   14383..14417
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        14706..14991
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(14993..15158,1..112)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
       61 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
      121 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
      181 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
      241 taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg tattaatgtt
      301 taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg tgccaccatg
      361 tacccatacg atgttccaga ttacgcttcg ccgaagaaaa agcgcaaggt cgaagcgtcc
      421 gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg ggccgtgatc
      481 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
      541 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
      601 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
      661 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
      721 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
      781 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
      841 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
      901 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
      961 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
     1021 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
     1081 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggcaacctg
     1141 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     1201 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     1261 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     1321 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     1381 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     1441 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     1501 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     1561 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     1621 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     1681 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     1741 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     1801 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     1861 gtggacaagg gcgcttcagc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     1921 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     1981 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     2041 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     2101 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     2161 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     2221 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     2281 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     2341 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     2401 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     2461 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     2521 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     2581 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     2641 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     2701 gaaatggcca gagagaacca aaccacccag aagggacaga agaacagccg cgagagaatg
     2761 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     2821 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     2881 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggacgccatc
     2941 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
     3001 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     3061 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
     3121 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     3181 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     3241 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
     3301 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     3361 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     3421 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     3481 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     3541 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
     3601 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
     3661 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
     3721 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
     3781 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
     3841 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
     3901 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
     3961 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
     4021 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
     4081 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
     4141 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
     4201 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
     4261 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
     4321 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
     4381 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
     4441 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
     4501 ctgtctcagc tgggaggcga cagccccaag aagaagagaa aggtggaggc cagcctctcg
     4561 aggccggacc ccgcgctggc cgcactcgga ggcggcggaa gcggtggcgg agggtcaggc
     4621 ggaggtggat ccatgaaatc ctgcggagtg tcgctcgcta ccgccgccgc tgccgccgcc
     4681 gctttcggtg atgaggaaaa gaaaatggcg gcgggaaaag cgagcggcga gagcgaggag
     4741 gcgtccccca gcctgacagc cgaggagagg gaggcgctcg gcggactgga cagccgcctc
     4801 tttgggttcg tgagatttca tgaagatggc gccaggacga aggccctact gggcaaggct
     4861 gttcgctgct atgaatctct aatcttaaaa gctgaaggaa aagtggagtc tgatttcttt
     4921 tgtcaattag gtcacttcaa cctcttattg gaagattatc caaaagcatt atctgcatac
     4981 cagaggtact acagtttaca gtctgactac tggaagaatg ctgccttttt atatggtctt
     5041 ggtttggtct acttccatta taatgcattt cagtgggcaa ttaaagcatt tcaggaggtg
     5101 ctttatgttg atcccagctt ttgtcgagcc aaggaaattc atttacgact tgggcttatg
     5161 ttcaaagtga acacagacta tgagtctagt ttaaagcatt ttcagttagc tttggttgac
     5221 tgtaatccct gcactttgtc caatgctgaa attcaatttc acattgccca cttatatgaa
     5281 acccagagga aatatcattc tgcaaaagaa gcttatgaac aacttttgca gacagagaat
     5341 ctttctgcac aagtaaaagc aactgtctta caacagttag gttggatgca tcacactgta
     5401 gatctcctgg gagataaagc caccaaggaa agctatgcta ttcagtatct ccaaaagtcc
     5461 ttggaagcag atcctaattc tggccagtcc tggtatttcc tcggaaggtg ctattcaagt
     5521 attgggaaag ttcaggatgc ctttatatct tacaggcagt ctattgataa atcagaagca
     5581 agtgcagata catggtgttc aataggtgtg ctatatcagc agcaaaatca gcccatggat
     5641 gctttacagg cctatatttg tgctgtacaa ttggaccatg gccatgctgc agcctggatg
     5701 gacctaggca ctctctatga atcctgcaac cagcctcagg atgccattaa atgctactta
     5761 aatgcaacta gaagcaaaag ttgtagtaat acctctgcac ttgcagcacg aattaagtat
     5821 ttacaggctc agttgtgtaa ccttccacaa ggtagtctac agaataaaac taaattactt
     5881 cctagtattg aggaggcgtg gagcctacca attcccgcag agcttacctc caggcagggt
     5941 gccatgaaca cagcacagca gaatacttct gacaattgga gtggtggaca tgctgtgtcg
     6001 catcctccag tacagcaaca agctcattca tggtgtttga caccacagaa attacagcat
     6061 ttggaacagc tccgggcaaa tagaaataat ttaaatccag cacagaaact gatgctggaa
     6121 cagctggaaa gtcagtttgt cttaatgcaa caacaccaaa tgagaccaac aggagttgca
     6181 caggtacgat ctactggaat tcctaatggg ccaacagctg actcatcact gcctacaaac
     6241 tcagtctctg gccagcagcc acagcttgct ctgaccagag tgcctagcgt ctctcagcct
     6301 ggagtccgtc ctgcctgccc tgggcagcct ttggccaatg gacccttttc tgcaggccat
     6361 gttccctgta gcacatcaag aacgctggga agtacagaca ctattttgat aggcaataat
     6421 catataacag gaagtggaag taatggaaac gtgccttacc tgcagcgaaa cgcactcact
     6481 ctacctcata accgcacaaa cctgaccagc agcgcagagg agccgtggaa aaaccaacta
     6541 tctaactcca ctcaggggct tcacaaaggt cagagttcac attcggcagg tcctaatggt
     6601 gaacgacctc tctcttccac tgggccttcc cagcatctcc aggcagctgg ctctggtatt
     6661 cagaatcaga acggacatcc caccctgcct agcaattcag taacacaggg ggctgctctc
     6721 aatcacctct cctctcacac tgctacctca ggtggacaac aaggcattac cttaaccaaa
     6781 gagagcaagc cttcaggaaa catattgacg gtgcctgaaa caagcaggca cactggagag
     6841 acacctaaca gcactgccag tgtcgaggga cttcctaatc atgtccatca gatgacggca
     6901 gatgctgttt gcagtcctag ccatggagat tctaagtcac caggtttact aagttcagac
     6961 aatcctcagc tctctgcctt gttgatggga aaagccaata acaatgtggg tactggaacc
     7021 tgtgacaaag tcaataacat ccacccagct gttcatacaa agactgataa ctctgttgcc
     7081 tcttcaccat cttcagccat ttcaacagca acaccttctc caaaatccac tgagcagaca
     7141 accacaaaca gtgttaccag ccttaacagc cctcacagtg ggctacacac aattaatgga
     7201 gaagggatgg aagaatctca gagccccatg aaaacagatc tgcttctggt taaccacaaa
     7261 cctagtccac agatcatacc atcaatgtct gtgtccatat accccagctc agcagaagtt
     7321 ctgaaggcat gcaggaatct aggtaaaaat ggcttatcta acagtagcat tttgttggat
     7381 aaatgtccac ctccaagacc accatcttca ccataccctc ccttgccaaa ggccaagttg
     7441 aatccaccta cacctagtat ttacttggaa aataaacgtg atgctttctt tcctccatta
     7501 catcaatttt gtacaaatcc gaacaaccct gttacagtaa tacgtggcct tgctggagct
     7561 cttaagttag acctgggact tttctctact aaaactttgg tggaagctaa caatgaacat
     7621 atggtagaag tgaggacaca gttgttgcag ccagcagatg aaaactggga tcccactgga
     7681 acaaagaaaa tctggcattg tgaaagtaat agatctcata ctacaattgc taaatatgca
     7741 cagtaccagg cctcctcatt ccaggaatca ttgagagaag aaaatgaaaa aagaagtcat
     7801 cataaagacc actcagatag tgaatctaca tcgtcagata attctgggag gaggaggaaa
     7861 ggacccttta aaaccataaa gtttgggacc aatattgacc tatctgatga caaaaagtgg
     7921 aagttgcagc tacatgagct gactaaactt cctgcttttg tgcgtgtcgt atcagcagga
     7981 aatcttctaa gccatgttgg tcataccata ttgggcatga acacagttca actatacatg
     8041 aaagttccag ggagcagaac accaggtcat caggaaaata acaacttctg ttcagttaac
     8101 ataaatattg gcccaggtga ctgtgaatgg tttgttgttc ctgaaggtta ctggggtgtt
     8161 ctgaatgact tctgtgaaaa aaataatttg aatttcctaa tgggttcttg gtggcccaat
     8221 cttgaagatc tttatgaagc aaatgttcca gtgtataggt ttattcagcg acctggagat
     8281 ttggtctgga taaatgcagg cactgttcat tgggttcagg ctattggctg gtgcaacaac
     8341 attgcttgga atgttggtcc acttacagcc tgccagtata aattggcagt ggaacggtac
     8401 gaatggaaca aattgcaaag tgtgaagtca atagtaccca tggttcatct ttcctggaat
     8461 atggcacgaa atatcaaggt ctcagatcca aagctttttg aaatgattaa gtattgtctt
     8521 ctaagaactc tgaagcaatg tcagacattg agggaagctc tcattgctgc aggaaaagag
     8581 attatatggc atgggcggac aaaagaagaa ccagctcatt actgtagcat ttgtgaagtg
     8641 gaggtttttg atctgctttt tgtcactaat gagagtaatt cacgaaagac ctacatagta
     8701 cattgccaag attgtgcacg aaaaacaagc ggaaacttgg aaaactttgt ggtgctagaa
     8761 cagtacaaaa tggaggacct gatgcaagtc tatgaccaat ttacattagc tcctccatta
     8821 ccatccgcct catcttgagc tagctaagaa ttcctagagc tcgctgatca gcctcgaact
     8881 cgaggttaat taatgagcgg ccgcatagat aactgatcca gtgtgctgga attaattcgc
     8941 tgtctgcgag ggccagctgt tggggtgagt actccctctc aaaagcgggc atgacttctg
     9001 cgctaagatt gtcagtttcc aaaaacgagg aggatttgat attcacctgg cccgcggtga
     9061 tgcctttgag ggtggccgcg tccatctggt cagaaaagac aatctttttg ttgtcaagct
     9121 tgaggtgtgg caggcttgag atctggccat acacttgagt gacaatgaca tccactttgc
     9181 ctttctctcc acaggtgtcc actcccaggt ccaactgcag gtcgagcatg catctagggc
     9241 ggccaattcc gcccctctcc cccccccccn tctccctccc ccccccctaa cgttactggc
     9301 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     9361 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     9421 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     9481 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     9541 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     9601 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     9661 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     9721 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     9781 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     9841 gcttgccaca acccgggata attcctgcag ccaatatgaa aaagcctgaa ctcaccgcga
     9901 cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct
     9961 cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc
    10021 gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat
    10081 cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct
    10141 attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc
    10201 ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc
    10261 agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg
    10321 atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca
    10381 ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc
    10441 ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg
    10501 gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg
    10561 tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact
    10621 tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca
    10681 ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg
    10741 cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa
    10801 tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg
    10861 gaaaccgacg ccccagcact cgtccgaggg caaaggaata gggatcaatt ctctagataa
    10921 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
    10981 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
    11041 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
    11101 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtcgag
    11161 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
    11221 cgacatacta gttaaaagtt ttgttacttt atagaagaaa ttttgagttt ttgttttttt
    11281 ttaataaata aataaacata aataaattgt ttgttgaatt tattattagt atgtaagtgt
    11341 aaatataata aaacttaata tctattcaaa ttaataaata aacctcgata tacagaccga
    11401 taaaacacat gcgtcaattt tacgcatgat tatctttaac gtacgtcaca atatgattat
    11461 ctttctaggg ttaatctagt atacgcgtat gcggccgctt aattaatcca gcttttgttc
    11521 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
    11581 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
    11641 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
    11701 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
    11761 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
    11821 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
    11881 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
    11941 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
    12001 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
    12061 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
    12121 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
    12181 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
    12241 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
    12301 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
    12361 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
    12421 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
    12481 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
    12541 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
    12601 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
    12661 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
    12721 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
    12781 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
    12841 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
    12901 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
    12961 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
    13021 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
    13081 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
    13141 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
    13201 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
    13261 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
    13321 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
    13381 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
    13441 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
    13501 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
    13561 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
    13621 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
    13681 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
    13741 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
    13801 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
    13861 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
    13921 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
    13981 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
    14041 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
    14101 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
    14161 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
    14221 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
    14281 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac
    14341 gactcactat agggcgaatt ggggcgcgcc attctagatt aaccctagaa agatagtctg
    14401 cgtaaaattg acgcatgcat tcttgaaata ttgctctctc tttctaaata gcgcgaatcc
    14461 gtcgctgtgc atttaggaca tctcagtcgc cgcttggagc tcccgtgagg cgtgcttgtc
    14521 aatgcggtaa gtgtcactga ttttgaacta taacgaccgc gtgagtcaaa atgacgcatg
    14581 attatctttt acgtgacttt taagatttaa ctcatacgat aattatattg ttatttcatg
    14641 ttctacttac gtgataactt attatatata tattttcttg ttatagatat caactagaat
    14701 gctagcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
    14761 ccattgacgt caatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
    14821 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
    14881 attgacgtca atgacggtaa atggcccgcc tggcattgtg cccagtacat gaccttatgg
    14941 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
    15001 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
    15061 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg gngncgcgcg
    15121 ccaggcgggg cggggcgggg cgaggggcgg ggcggggc
//
