LOCUS       sequence-230194-        9176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR-Cas9 for miR-124-1/2, 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-124-1/2-5'
FEATURES             Location/Qualifiers
     source          1..9176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        complement(76..151)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(173..421)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(231..250)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(401..421)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(483..1071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(563..582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1242..2033,2034..2102))
                     /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     1865..1884
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2103..2207)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2275..2293
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2331..2353)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2453..2472
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2489..2944)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2626..2647)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2838..2857
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3019..3159
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3030..3159
                     /label=AAV2 ITR
     polyA_signal    3168..3375
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3364..3381
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3409..4008)
                     /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(3492..3512)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3989..4008
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(4009..4062)
                     /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             complement(4078..4125)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(4126..8226)
                     /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             complement(4126..8226)
                     /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             complement(8251..8271)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8278..8343)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      8343..8352
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          8364..8591
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     promoter        complement(8592..8868)
                     /label=chicken beta-actin promoter
     enhancer        8870..9155
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ctgcagaatt ggcgcacgcg ctaaaaacgg actagcctta ttttaacttg ctatttctag
       61 ctctaaaaca aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct
      121 tattttaact tgctatttct agctctaaaa ctgttcacag cggaccttga tcggtgtttc
      181 gtcctttcca caagatatat aaagccaaga aatcgaaata ctttcaagtt acggtaagca
      241 tatgatagtc cattttaaaa cataatttta aaactgcaaa ctacccaaga aattattact
      301 ttctacgtca cgtattttgt actaatatct ttgtgtttac agtcaaatta attccaatta
      361 tctctctaac agccttgtat cgtatatgca aatatgaagg aatcatggga aataggccct
      421 cacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      481 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      541 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      601 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      661 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      721 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      781 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      841 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      901 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      961 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     1021 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     1081 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     1141 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     1201 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     1261 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1321 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1381 cggcttccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1441 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1501 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1561 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1621 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1681 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1741 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1801 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1861 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1921 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1981 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     2041 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     2101 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     2161 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     2221 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     2281 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     2341 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     2401 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     2461 gagcagattg tactgagagt gcaccataaa attgtaaacg ttaatatttt gttaaaattc
     2521 gcgttaaatt tttgttaaat cagctcattt tttaaccaat agaccgaaat cggcaaaatc
     2581 ccttataaat caaaagaata gcccgagata gagttgagtg ttgttccagt ttggaacaag
     2641 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2701 gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa
     2761 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2821 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt
     2881 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     2941 gcgtactatg gttgctttga cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     3001 aataccgcat caggcgcccc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc
     3061 gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
     3121 gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgctcc ccagcatgcc
     3181 tgctattctc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc cccagaatag
     3241 aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg
     3301 gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc
     3361 aactagaagg cacagtcgag gctgatcagc gagctctagt tagaattctc aggcaccggg
     3421 cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt
     3481 gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg
     3541 caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt
     3601 gccctggtgg tcgggcgaga ctccgacggt ggccaggaac cacgcgggct ccttgggccg
     3661 gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc
     3721 ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt
     3781 ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac
     3841 gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt ccggatcgac
     3901 ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg
     3961 gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcattg ggccaggatt
     4021 ctcctcgacg tcaccgcatg ttagcagact tcctctgccc tctccactgc cgaattcctt
     4081 tttctttttt gcctggccgg cctttttcgt ggccgccggc cttttgtcgc ctcccagctg
     4141 agacaggtcg atccgtgtct cgtacaggcc ggtgatgctc tggtggatca gggtggcgtc
     4201 cagcacctct ttggtgctgg tgtacctctt ccggtcgatg gtggtgtcaa agtacttgaa
     4261 ggcggcaggg gctcccagat tggtcagggt aaacaggtgg atgatattct cggcctgctc
     4321 tctgatgggc ttatcccggt gcttgttgta ggcggacagc actttgtcca gattagcgtc
     4381 ggccaggatc actctcttgg agaactcgct gatctgctcg atgatctcgt ccaggtagtg
     4441 cttgtgctgt tccacaaaca gctgtttctg ctcattatcc tcgggggagc ccttcagctt
     4501 ctcatagtgg ctggccaggt acaggaagtt cacatatttg gagggcaggg ccagttcgtt
     4561 tcccttctgc agttcgccgg cagaggccag cattctcttc cggccgtttt ccagctcgaa
     4621 cagggagtac ttaggcagct tgatgatcag gtcctttttc acttctttgt agcccttggc
     4681 ttccagaaag tcgatgggat tcttctcgaa gctgcttctt tccatgatgg tgatccccag
     4741 cagctctttc acactcttca gtttcttgga cttgcccttt tccactttgg ccaccaccag
     4801 cacagaatag gccacggtgg ggctgtcgaa gccgccgtac ttcttagggt cccagtcctt
     4861 ctttctggcg atcagcttat cgctgttcct cttgggcagg atagactctt tgctgaagcc
     4921 gcctgtctgc acctcggtct ttttcacgat attcacttgg ggcatgctca gcactttccg
     4981 cacggtggca aaatcccggc ccttatccca cacgatctcc ccggtttcgc cgtttgtctc
     5041 gatcagaggc cgcttccgga tctcgccgtt ggccagggta atctcggtct tgaaaaagtt
     5101 catgatgttg ctgtagaaga agtacttggc ggtagccttg ccgatttcct gctcgctctt
     5161 ggcgatcatc ttccgcacgt cgtacacctt gtagtcgccg tacacgaact cgctttccag
     5221 cttagggtac tttttgatca gggcggttcc cacgacggcg ttcaggtagg cgtcgtgggc
     5281 gtggtggtag ttgttgatct cgcgcacttt gtaaaactgg aaatccttcc ggaaatcgga
     5341 caccagcttg gacttcaggg tgatcacttt cacttcccgg atcagcttgt cattctcgtc
     5401 gtacttagtg ttcatccggg agtccaggat ctgtgccacg tgctttgtga tctgccgggt
     5461 ttccaccagc tgtctcttga tgaagccggc cttatccagt tcgctcaggc cgcctctctc
     5521 ggccttggtc agattgtcga actttctctg ggtaatcagc ttggcgttca gcagctgccg
     5581 ccagtagttc ttcatcttct tcacgacctc ttcggagggc acgttgtcgc tcttgccccg
     5641 gttcttgtcg cttctggtca gcaccttgtt gtcgatggag tcgtccttca gaaagctctg
     5701 aggcacgata tggtccacat cgtagtcgga cagccggttg atgtccagtt cctggtccac
     5761 gtacatatcc cgcccattct gcaggtagta caggtacagc ttctcgttct gcagctgggt
     5821 gttttccacg gggtgttctt tcaggatctg gctgcccagc tctttgatgc cctcttcgat
     5881 ccgcttcatt ctctcgcggc tgttcttctg tcccttctgg gtggtctggt tctctctggc
     5941 catttcgatc acgatgttct cgggcttgtg ccggcccatc actttcacga gctcgtccac
     6001 caccttcact gtctgcagga tgcccttctt aatggcgggg ctgccggcca gattggcaat
     6061 gtgctcgtgc aggctatcgc cctggccgga cacctgggct ttctggatgt cctctttaaa
     6121 ggtcaggctg tcgtcgtgga tcagctgcat gaagtttctg ttggcgaagc cgtcggactt
     6181 caggaaatcc aggattgtct tgccggactg cttgtcccgg atgccgttga tcagcttccg
     6241 gctcagcctg ccccagccgg tgtatctccg ccgcttcagc tgcttcatca ctttgtcgtc
     6301 gaacaggtgg gcataggttt tcagccgttc ctcgatcatc tctctgtcct caaacagtgt
     6361 cagggtcagc acgatatctt ccagaatgtc ctcgttttcc tcattgtcca ggaagtcctt
     6421 gtccttgata attttcagca gatcgtggta tgtgcccagg gaggcgttga accgatcttc
     6481 cacgccggag atttccacgg agtcgaagca ctcgattttc ttgaagtagt cctctttcag
     6541 ctgcttcacg gtcactttcc ggttggtctt gaacagcagg tccacgatgg cctttttctg
     6601 ctcgccgctc aggaaggcgg gctttctcat tccctcggtc acgtatttca ctttggtcag
     6661 ctcgttatac acggtgaagt actcgtacag caggctgtgc ttgggcagca ccttctcgtt
     6721 gggcaggttc ttatcgaagt tggtcatccg ctcgatgaag ctctgggcgg aagcgccctt
     6781 gtccaccact tcctcgaagt tccagggggt gatggtttcc tcgctctttc tggtcatcca
     6841 ggcgaatctg ctgtttcccc tggccagagg gcccacgtag taggggatgc ggaaggtcag
     6901 gatcttctcg atcttttccc ggttgtcctt caggaatggg taaaaatctt cctgccgccg
     6961 cagaatggcg tgcagctctc ccaggtggat ctggtggggg atgctgccgt tgtcgaaggt
     7021 ccgctgcttc cgcagcaggt cctctctgtt cagcttcacg agcagttcct cggtgccgtc
     7081 catcttttcc aggatgggct tgatgaactt gtagaactct tcctggctgg ctccgccgtc
     7141 aatgtagccg gcgtagccgt tcttgctctg gtcgaagaaa atctctttgt acttctcagg
     7201 cagctgctgc cgcacgagag ctttcagcag ggtcaggtcc tggtggtgct cgtcgtatct
     7261 cttgatcata gaggcgctca ggggggcctt ggtgatctcg gtgttcactc tcaggatgtc
     7321 gctcagcagg atggcgtcgg acaggttctt ggcggccaga aacaggtcgg cgtactggtc
     7381 gccgatctgg gccagcaggt tgtccaggtc gtcgtcgtag gtgtccttgc tcagctgcag
     7441 tttggcatcc tcggccaggt cgaagttgct cttgaagttg ggggtcaggc ccaggctcag
     7501 ggcaatcagg tttccgaaca ggccattctt cttctcgccg ggcagctggg cgatcagatt
     7561 ttccagccgt ctgctcttgc tcagtctggc agacaggatg gccttggcgt ccacgccgct
     7621 ggcgttgatg gggttttcct cgaacagctg gttgtaggtc tgcaccagct ggatgaacag
     7681 cttgtccacg tcgctgttgt cggggttcag gtcgccctcg atcaggaagt ggccccggaa
     7741 cttgatcatg tgggccaggg ccagatagat cagccgcagg tcggccttgt cggtgctgtc
     7801 caccagtttc tttctcaggt ggtagatggt ggggtacttc tcgtggtagg ccacctcgtc
     7861 cacgatgttg ccgaagatgg ggtgccgctc gtgcttctta tcctcttcca ccaggaagga
     7921 ctcttccagt ctgtggaaga agctgtcgtc caccttggcc atctcgttgc tgaagatctc
     7981 ttgcagatag cagatccggt tcttccgtct ggtgtatctt cttctggcgg ttctcttcag
     8041 ccgggtggcc tcggctgttt cgccgctgtc gaacagcagg gctccgatca ggttcttctt
     8101 gatgctgtgc cggtcggtgt tgcccagcac cttgaatttc ttgctgggca ccttgtactc
     8161 gtcggtgatc acggcccagc ccacagagtt ggtgccgatg tccaggccga tgctgtactt
     8221 cttgtcggct gctgggactc cgtggatacc gaccttccgc ttcttctttg gggccatctt
     8281 atcgtcatcg tctttgtaat caatatcatg atccttgtag tctccgtcgt ggtccttata
     8341 gtccatggtg gcaccggtcc aacctgaaaa aaagtgattt caggcaggtg ctccaggtaa
     8401 ttaaacatta ataccccacc aaccaaccat cccttaaacc cttacctctt gctcagctaa
     8461 ttacagcccg gaggagaagg gccgtcccgc ccgctcacct gtgggagtaa cgcggtcagt
     8521 cagagccggg gcgggcggcg cgaggcggcg gcggagcggg gcacggggcg aaggcagcgc
     8581 gcagcgactc ccgcccgccg cgcgcttcgc tttttatagg gccgccgccg ccgccgcctc
     8641 gccataaaag gaaactttcg gagcgcgccg ctctgattgg ctgccgccgc acctctccgc
     8701 ctcgccccgc cccgcccctc gccccgcccc gccccgcctg gcgcgcgccc cccccccccc
     8761 ccgcccccat cgctgcacaa aataattaaa aaataaataa atacaaaatt gggggtgggg
     8821 agggggggga gatggggaga gtgaagcaga acgtggggct cacctcgacc atggtaatag
     8881 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     8941 cacaatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     9001 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     9061 aatggaaagt ccctattggc gttactattg acgtcaatgg gcgggggtcg ttgggcggtc
     9121 agccaggcgg gccatttacc gtaagttatg taacgggtac ctctagagcc atttgt
//
