LOCUS       sequence-252314-        8446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sgRNA targeting mouse Birc6 and Cas9 nickase in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8446)
  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 8446)
  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)-Birc6-L
FEATURES             Location/Qualifiers
     source          1..8446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          161..388
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             409..435
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             439..459
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(469..492,493..495,496..4569)
                     /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             469..4569
                     /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             4573..4593
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4630..4647)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4636..4843
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4852..4992
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4852..4981
                     /label=AAV2 ITR
     rep_origin      5067..5522
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5154..5173)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5364..5385
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5539..5558)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5658..5680
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5718..5736)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5804..5908
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5909..5977,5978..6769)
                     /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     complement(6127..6146)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6940..7528
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7429..7448
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7590..7838
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7590..7610
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7761..7780
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        8042..8327
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(8329..8446,1..160)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg
       61 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
      121 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
      181 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
      241 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagctgag
      301 caagaggtaa gggtttaagg gatggttggt tggtggggta ttaatgttta attacctgga
      361 gcacctgcct gaaatcactt tttttcaggt tggaccggtg ccaccatgta cccatacgat
      421 gttccagatt acgcttcgcc gaagaaaaag cgcaaggtcg aagcgtccga caagaagtac
      481 agcatcggcc tggccatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
      541 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
      601 aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag
      661 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
      721 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
      781 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
      841 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
      901 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
      961 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
     1021 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
     1081 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg
     1141 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gcaacctgat tgccctgagc
     1201 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
     1261 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     1321 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
     1381 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac
     1441 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
     1501 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc
     1561 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
     1621 accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc
     1681 gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
     1741 caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
     1801 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
     1861 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
     1921 gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
     1981 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc
     2041 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
     2101 aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
     2161 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
     2221 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
     2281 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
     2341 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
     2401 aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag
     2461 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
     2521 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
     2581 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
     2641 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
     2701 gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga
     2761 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
     2821 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
     2881 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
     2941 caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc
     3001 tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc
     3061 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag
     3121 ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga
     3181 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
     3241 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
     3301 aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
     3361 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
     3421 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa
     3481 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
     3541 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
     3601 ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca
     3661 aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa
     3721 gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
     3781 agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac
     3841 tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg
     3901 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg
     3961 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
     4021 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
     4081 ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa
     4141 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
     4201 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac
     4261 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
     4321 aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag
     4381 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
     4441 tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc
     4501 accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
     4561 ggaggcgaca gccccaagaa gaagagaaag gtggaggcca gctaagaatt cctagagctc
     4621 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     4681 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     4741 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     4801 gcaaggggga ggattgggaa gagaatagca ggcatgctgg ggagcggccg caggaacccc
     4861 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     4921 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     4981 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     5041 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     5101 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     5161 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5221 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5281 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5341 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     5401 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     5461 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     5521 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     5581 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     5641 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     5701 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     5761 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5821 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5881 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5941 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     6001 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     6061 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     6121 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     6181 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     6241 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     6301 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     6361 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     6421 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     6481 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     6541 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6601 taaatctgga gccggtgagc gtggaagccg cggtatcatt gcagcactgg ggccagatgg
     6661 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6721 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6781 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6841 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6901 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6961 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     7021 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7081 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7141 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7201 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7261 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7321 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7381 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7441 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7501 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7561 ggccttttgc tggccttttg ctcacatgtg agggcctatt tcccatgatt ccttcatatt
     7621 tgcatatacg atacaaggct gttagagaga taattggaat taatttgact gtaaacacaa
     7681 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt
     7741 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt
     7801 tcttggcttt atatatcttg tggaaaggac gaaacaccgt gtggatttcg tagacacaag
     7861 tttaagagct atgctggaaa cagcatagca agtttaaata aggctagtcc gttatcaact
     7921 tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag ctagaaatag caagttaaaa
     7981 taaggctagt ccgtttttag cgcgtgcgcc aattctgcag acaaatggct ctagaggtac
     8041 ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     8101 tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     8161 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     8221 acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc ttatgggact
     8281 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc
     8341 ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat
     8401 ttatttttta attattttgt gcagcgatgg gggcgggggg gggggg
//
