LOCUS       sequence-252308-        8443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sgRNA targeting mouse Dhx15 and Cas9 nickase in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8443)
  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 8443)
  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: px335-sgRNA-EF(Cas9n)-Dhx15-L
FEATURES             Location/Qualifiers
     source          1..8443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          134..361
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             382..408
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             412..432
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(442..465,466..468,469..4542)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             442..4542
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4546..4566
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4603..4620)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4609..4816
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4825..4965
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4825..4954
                     /label=AAV2 ITR
     rep_origin      5040..5495
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5127..5146)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5337..5358
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5512..5531)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5631..5653
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5691..5709)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5777..5881
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5882..5950,5951..6742)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESSRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6100..6119)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6913..7501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7402..7421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7563..7811
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7563..7583
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7734..7753
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        8015..8300
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(8302..8443,1..133)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
       61 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      121 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
      181 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
      241 cggcccttct cctccgggct gtaattagct gagcaagagg taagggttta agggatggtt
      301 ggttggtggg gtattaatgt ttaattacct ggagcacctg cctgaaatca ctttttttca
      361 ggttggaccg gtgccaccat gtacccatac gatgttccag attacgcttc gccgaagaaa
      421 aagcgcaagg tcgaagcgtc cgacaagaag tacagcatcg gcctggccat cggcaccaac
      481 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
      541 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
      601 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
      661 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
      721 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
      781 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
      841 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
      901 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
      961 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     1021 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     1081 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     1141 aatggcctgt tcggcaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     1201 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     1261 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     1321 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     1381 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
     1441 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     1501 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag
     1561 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     1621 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
     1681 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
     1741 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     1801 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     1861 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg
     1921 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     1981 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg
     2041 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag
     2101 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     2161 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     2221 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     2281 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     2341 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
     2401 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     2461 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     2521 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     2581 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     2641 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
     2701 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     2761 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     2821 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     2881 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     2941 gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac
     3001 aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
     3061 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag
     3121 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     3181 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     3241 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa
     3301 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     3361 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     3421 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     3481 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     3541 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     3601 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
     3661 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
     3721 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
     3781 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     3841 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     3901 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
     3961 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     4021 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     4081 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     4141 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
     4201 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc
     4261 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac
     4321 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc
     4381 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
     4441 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
     4501 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acagccccaa gaagaagaga
     4561 aaggtggagg ccagctaaga attcctagag ctcgctgatc agcctcgact gtgccttcta
     4621 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     4681 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     4741 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata
     4801 gcaggcatgc tggggagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     4861 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4921 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     4981 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     5041 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5101 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5161 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5221 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     5281 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5341 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     5401 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5461 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     5521 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     5581 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     5641 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     5701 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     5761 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5821 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5881 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5941 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6001 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttctcgccc
     6061 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6121 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6181 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6241 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6301 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6361 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6421 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6481 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6541 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag
     6601 ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6661 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6721 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6781 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6841 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6901 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6961 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     7021 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     7081 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     7141 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     7201 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7261 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7321 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7381 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7441 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7501 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7561 gtgagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
     7621 agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
     7681 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
     7741 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc tagtggaaag
     7801 gacgaaacac cgtcagtact gtgaatattc ctgtttaaga gctatgctgg aaacagcata
     7861 gcaagtttaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     7921 ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtttt tagcgcgtgc
     7981 gccaattctg cagacaaatg gctctagagg tacccgttac ataacttacg gtaaatggcc
     8041 cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg ccaataggga
     8101 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     8161 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     8221 ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     8281 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
     8341 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     8401 tgggggcggg gggggggggn cgcgcgccag gcggggcggg gcg
//
