LOCUS       sequence-230197-        9176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR-Cas9 for miR-124-3, 5'.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9176)
  AUTHORS   Kutsche LK, Gysi DM, Fallmann J, Lenk K, Petri R, Swiersy A, Klapper
            SD, Pircs K, Khattak S, Stadler PF, Jakobsson J, Nowick K, Busskamp 
            V
  TITLE     Combined Experimental and System-Level Analyses Reveal the Complex 
            Regulatory Network of miR-124 during Human Neurogenesis.
  JOURNAL   Cell Syst. 2018 Sep 28. pii: S2405-4712(18)30358-2. doi: 
            10.1016/j.cels.2018.08.011.
  PUBMED    30292704
REFERENCE   2  (bases 1 to 9176)
  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: "Cell Syst. 
            2018 Sep 28. pii: S2405-4712(18)30358-2. doi: 
            10.1016/j.cels.2018.08.011."
COMMENT     Sequence Label: pSpCas9(BB)-2A-Puro (PX459)+gRNA miR-124-3-5'
FEATURES             Location/Qualifiers
     source          1..9176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        91..339
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     91..111
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     262..281
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        360..435
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        532..817
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        819..1096
                     /label=chicken beta-actin promoter
     intron          1097..1324
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1336..1345
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1345..1410
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1417..1437
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1462..5562
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1462..5562
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5563..5610
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5626..5679
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5680..6279
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5680..5699)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6176..6196
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6307..6324)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6313..6520
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6529..6669
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6529..6658
                     /label=AAV2 ITR
     rep_origin      6744..7199
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6831..6850)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7041..7062
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7216..7235)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7335..7357
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7395..7413)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7481..7585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7586..7654,7655..8446)
                     /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(7804..7823)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8617..9176,1..29)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9106..9125
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
       61 tggccttttg ctggcctttt gctcacatgt gagggcctat ttcccatgat tccttcatat
      121 ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca
      181 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
      241 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
      301 ttcttggctt tatatatctt gtggaaagga cgaaacaccg tctatacaat taaggcacgg
      361 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
      421 gcaccgagtc ggtgcttttt tgttttagag ctagaaatag caagttaaaa taaggctagt
      481 ccgtttttag cgcgtgcgcc aattctgcag acaaatggct ctagaggtac ccgttacata
      541 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      601 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      661 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      721 cggtaaatgg cccgcctggc attgtgccca gtacatgacc ttatgggact ttcctacttg
      781 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
      841 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
      901 attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccag gcggggcggg
      961 gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg
     1021 cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg
     1081 aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc
     1141 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
     1201 ggacggccct tctcctccgg gctgtaatta gctgagcaag aggtaagggt ttaagggatg
     1261 gttggttggt ggggtattaa tgtttaatta cctggagcac ctgcctgaaa tcactttttt
     1321 tcaggttgga ccggtgccac catggactat aaggaccacg acggagacta caaggatcat
     1381 gatattgatt acaaagacga tgacgataag atggccccaa agaagaagcg gaaggtcggt
     1441 atccacggag tcccagcagc cgacaagaag tacagcatcg gcctggacat cggcaccaac
     1501 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
     1561 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
     1621 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
     1681 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
     1741 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
     1801 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
     1861 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     1921 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     1981 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     2041 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     2101 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     2161 aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     2221 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     2281 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     2341 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     2401 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
     2461 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     2521 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag
     2581 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     2641 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
     2701 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
     2761 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     2821 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     2881 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg
     2941 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     3001 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg
     3061 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag
     3121 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     3181 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     3241 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     3301 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     3361 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
     3421 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     3481 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     3541 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     3601 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     3661 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
     3721 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     3781 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     3841 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     3901 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     3961 gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac
     4021 aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
     4081 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag
     4141 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     4201 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     4261 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa
     4321 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     4381 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     4441 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     4501 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     4561 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     4621 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
     4681 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
     4741 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
     4801 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     4861 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     4921 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
     4981 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     5041 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     5101 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     5161 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
     5221 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc
     5281 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac
     5341 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc
     5401 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
     5461 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
     5521 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg
     5581 aaaaaggccg gccaggcaaa aaagaaaaag gaattcggca gtggagaggg cagaggaagt
     5641 ctgctaacat gcggtgacgt cgaggagaat cctggcccaa tgaccgagta caagcccacg
     5701 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     5761 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     5821 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     5881 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     5941 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     6001 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     6061 gtcggagtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     6121 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     6181 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     6241 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag aattctaact agagctcgct
     6301 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     6361 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     6421 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     6481 agggggagga ttgggaagag aatagcaggc atgctgggga gcggccgcag gaacccctag
     6541 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     6601 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     6661 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     6721 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     6781 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     6841 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     6901 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     6961 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     7021 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     7081 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     7141 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     7201 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     7261 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     7321 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     7381 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     7441 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     7501 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     7561 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     7621 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     7681 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     7741 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     7801 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     7861 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     7921 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     7981 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     8041 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     8101 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     8161 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     8221 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     8281 atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa
     8341 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     8401 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     8461 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     8521 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     8581 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     8641 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     8701 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     8761 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     8821 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     8881 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     8941 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     9001 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     9061 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     9121 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgat
//
