LOCUS       sequence-435357-       10963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid contains hCas9 and two gRNAs that can excise the 
            genomic area around the mouse mdx mutation in dystrophin's exon 23.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10963)
  AUTHORS   Lenardic A, Domenig SA, Zvick J, Bundschuh N, Tarnowska-Sengul M, 
            Furrer R, Noe FJ, Trautmann CLL, Ghosh A, Bacchin G, Gjonlleshaj P, 
            Qabrati X, Masschelein E, De Bock K, Handschin C, Bar-Nur O
  TITLE     Generation of allogenic and xenogeneic functional muscle stem cells 
            for intramuscular transplantation.
  JOURNAL   J Clin Invest. 2024 May 7:e166998. doi: 10.1172/JCI166998.
  PUBMED    38713532
REFERENCE   2  (bases 1 to 10963)
  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: "J Clin 
            Invest. 2024 May 7:e166998. doi: 10.1172/JCI166998."
COMMENT     Sequence Label: pRP[CRISPR]-EGFP/Puro-hCas9-U6>(long left)-U6>...
FEATURES             Location/Qualifiers
     source          1..10963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        271..346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        361..609
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        631..706
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        726..1011
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1013..1290
                     /label=chicken beta-actin promoter
     intron          1291..1518
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1530..1539
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1539..1604
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1611..1631
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1656..5756
                     /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             1656..5756
                     /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             5757..5804
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5832..5849)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5838..6045
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        6060..6439
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6440..6643
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6594..6614
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6670..6679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6676..6678,6679..6681,6682..7392)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6676..6678,6679..6681,6682..7392)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6982..7001)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7329..7350
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7393..7992
                     /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(7393..7412)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7889..7909
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8005..8126
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8016..8035)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8070..8089
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   8226..8366
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   8226..8355
                     /label=AAV2 ITR
     rep_origin      8441..8896
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8528..8547)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8738..8759
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8913..8932)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9032..9054
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9092..9110)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9178..9282
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9283..9351,9352..10143)
                     /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(9501..9520)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10314..10902
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10803..10822
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg tctttgaaag agcaataaaa gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttggccggcc
      361 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
      421 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      481 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      541 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      601 cgaaacaccg atttcaggta agccgaggtt gttttagagc tagaaatagc aagttaaaat
      661 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgtctagag
      721 gtacccgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      781 ccattgacgt caatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      841 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      901 attgacgtca atgacggtaa atggcccgcc tggcattgtg cccagtacat gaccttatgg
      961 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
     1021 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
     1081 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg
     1141 ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca
     1201 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg
     1261 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg
     1321 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
     1381 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagctgag caagaggtaa
     1441 gggtttaagg gatggttggt tggtggggta ttaatgttta attacctgga gcacctgcct
     1501 gaaatcactt tttttcaggt tggaccggtg ccaccatgga ctataaggac cacgacggag
     1561 actacaagga tcatgatatt gattacaaag acgatgacga taagatggcc ccaaagaaga
     1621 agcggaaggt cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg
     1681 acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
     1741 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag
     1801 ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa
     1861 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
     1921 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     1981 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     2041 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     2101 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     2161 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     2221 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
     2281 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
     2341 ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc
     2401 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     2461 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     2521 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     2581 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     2641 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     2701 ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg
     2761 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
     2821 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
     2881 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
     2941 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
     3001 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
     3061 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga
     3121 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
     3181 agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg
     3241 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg
     3301 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
     3361 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
     3421 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
     3481 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
     3541 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
     3601 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
     3661 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
     3721 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
     3781 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
     3841 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
     3901 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
     3961 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
     4021 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
     4081 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
     4141 tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg
     4201 actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg
     4261 tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca
     4321 agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
     4381 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
     4441 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga
     4501 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
     4561 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
     4621 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
     4681 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
     4741 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
     4801 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg
     4861 gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc
     4921 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
     4981 tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga
     5041 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
     5101 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
     5161 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
     5221 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
     5281 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
     5341 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
     5401 aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca
     5461 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag
     5521 tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca
     5581 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
     5641 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
     5701 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaaaa
     5761 ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaagtaagaa ttcctagagc
     5821 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     5881 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     5941 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     6001 cagcaagggg gaggattggg aagagaatag caggcatgct ggggagcggc cgccgcgttg
     6061 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     6121 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     6181 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     6241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     6301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     6361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     6421 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     6481 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     6541 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     6601 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
     6661 acccactgcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     6721 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     6781 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     6841 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     6901 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     6961 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     7021 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     7081 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     7141 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     7201 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     7261 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     7321 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     7381 gagctgtaca agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     7441 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     7501 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     7561 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     7621 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     7681 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     7741 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     7801 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     7861 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     7921 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     7981 cccggtgcct gagagcagac atgataagat acattgatga gtttggacaa accacaacta
     8041 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     8101 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     8161 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtagcg
     8221 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     8281 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     8341 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     8401 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     8461 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     8521 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     8581 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     8641 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     8701 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     8761 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     8821 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     8881 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     8941 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     9001 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     9061 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     9121 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     9181 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     9241 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     9301 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     9361 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     9421 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     9481 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     9541 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     9601 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     9661 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     9721 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     9781 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     9841 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     9901 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     9961 tggtttattg ctgataaatc tggagccggt gagcgtggaa gccgcggtat cattgcagca
    10021 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
    10081 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
    10141 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
    10201 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
    10261 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
    10321 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
    10381 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
    10441 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
    10501 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
    10561 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
    10621 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
    10681 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
    10741 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
    10801 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
    10861 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
    10921 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
