LOCUS       sequence-287975-       13977 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses SpCas9, and cloning backbone for sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 13977)
  AUTHORS   Xie Y, Wang D, Lan F, Wei G, Ni T, Chai R, Liu D, Hu S, Li M, Li D, 
            Wang H, Wang Y
  TITLE     An episomal vector-based CRISPR/Cas9 system for highly efficient 
            gene knockout in human pluripotent stem cells.
  JOURNAL   Sci Rep. 2017 May 24;7(1):2320. doi: 10.1038/s41598-017-02456-y.
  PUBMED    28539611
REFERENCE   2  (bases 1 to 13977)
  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 Rep."; 
            date: "2017-05-24"; volume: "7(1):2320. doi"; pages: " 
            10.1038/s41598-017-02456-y"
COMMENT     Sequence Label: epiCRISPRe
FEATURES             Location/Qualifiers
     source          1..13977
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(1155..1173)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1241..1345
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1346..1414,1415..2206)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1564..1583)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2377..2965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2866..2885
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3154..3174)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3270..3518
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3270..3290
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3441..3460
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3539..3614
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    3662..3779
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3848..4059
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4079..4088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4088..4111
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4118..4138
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4163..6700,6701..6703,6704..7165,7166..7168,7169..7336,
                     7337..7339,7340..8263)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Slaymaker et al., 2016)"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8264..8311
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8327..8383
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             8384..8980
                     /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(8384..8403)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8880..8900
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             9008..9061
                     /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             9092..9754
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
     polyA_signal    9874..10008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9898..9917)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     9952..9971
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(10151..10173)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      10733..12522
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
ORIGIN
        1 tgcccctcct gctcctgccc ctcctgctcc tgcccctcct gcccctcctc ctgctcctgc
       61 ccctcctcct gctcctgccc ctcctcctcc tccccctcct ccccctcctc ctcctccccc
      121 tccycccccc ccyccccctc cyccccctcc ycccccyccy ccyccycctc cccctccycc
      181 ccctcctcct cctccycccc ctcctcctcc yccycccccy cctccccctc ctccccctcc
      241 tccycctcct ccccctcctc cccctcctcc tccycctccc cctcctcctc ctcctccccc
      301 tcctccccct cctccccctc ctcctcctcc tccccctcct cctgctcctg cccctcctgc
      361 ccctcctscc cctcctgccc ctcctcctgc tcctgcccct cctcctgctc ctgcccctcc
      421 tgctcctgcc cctcccgctc ctgctcctgc tcctgttcca ccgtgggtcc ctttgcagcc
      481 aatgcaactt ggacgttttt ggggtctccg gacaccatct ctatgtcttg gccctgatcc
      541 tgagccgccc ggggctcctg gtcttccgcc tcctcgtcct cgtcctcttc cccgtcctcg
      601 tccatggtta tcaccccctc ttctttgagg tccactgccg ccggagcctt ctggtccaga
      661 tgtgtctccc ttctctccta ggccatttcc aggtcctgta cctggcccct cgtcagacat
      721 gattcacact aaaagagatc aatagacatc tttattagac gacgctcagt gaatacaggg
      781 agtgcagact cctgccccct ccaacagccc ccccaccctc atccccttca tggtcgctgt
      841 cagacagatc caggtctgaa aattccccat cctccgaacc atcctcgtcc tcatcaccaa
      901 ttactcgcag cccggaaaac tcccgctgaa catcctcaag atttgcgtcc tgagcctcaa
      961 gccaggcctc aaattcctcg tccccctttt tgctggacgg tagggatggg gattctcggg
     1021 acccctcctc ttcctcttca aggtcaccag acagagatgc tactggggca acggaagaaa
     1081 agctgggtgc ggcctgtgag gatcagctta tcgatgataa gctgtcaaac atgagaattc
     1141 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     1201 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1261 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1321 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1381 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1441 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1501 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1561 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     1621 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     1681 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1741 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1801 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1861 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     1921 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     1981 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2041 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2101 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2161 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2221 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2281 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2341 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2401 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2461 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2521 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2581 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2641 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2701 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2761 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2821 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2881 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2941 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3001 cttttgctgg ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc
     3061 ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat
     3121 ccagcctcgc gtcgaactag atgatccggc tgtggaatgt gtgtcagtta gggtgtggaa
     3181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcattatggt acctttcaga
     3241 cccacctccc aaccccgagg ggacccagag agggcctatt tcccatgatt ccttcatatt
     3301 tgcatatacg atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa
     3361 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt
     3421 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt
     3481 tcttggcttt atatatcttg tggaaaggac gaaacaccga gaagagcgat gctcttcggt
     3541 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
     3601 caccgagtcg gtgctttttt gaattctaga tcttgagaca aatggcagta ttcatccaca
     3661 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     3721 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
     3781 gggtttatta cagggacagc agagatccac tttggcgccg gctcgagtgg ctccggtgcc
     3841 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     3901 aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     3961 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     4021 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg tgtcgtgacg cgggatccgc
     4081 caccatggat tacaaagacg atgacgataa gatggcccca aagaagaagc ggaaggtcgg
     4141 tatccacgga gtcccagcag ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa
     4201 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
     4261 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
     4321 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
     4381 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     4441 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     4501 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     4561 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     4621 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     4681 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     4741 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     4801 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     4861 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     4921 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     4981 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     5041 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     5101 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     5161 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     5221 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     5281 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     5341 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     5401 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     5461 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     5521 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     5581 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     5641 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     5701 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     5761 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     5821 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     5881 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     5941 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     6001 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     6061 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     6121 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     6181 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     6241 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     6301 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     6361 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     6421 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     6481 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     6541 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     6601 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     6661 cgactacgat gtggaccata tcgtgcctca gagctttctg gccgacgact ccatcgacaa
     6721 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     6781 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     6841 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     6901 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     6961 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     7021 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     7081 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     7141 aaccgccctg atcaaaaagt accctgcgct ggaaagcgag ttcgtgtacg gcgactacaa
     7201 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     7261 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     7321 cggcgagatc cggaaggcgc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     7381 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     7441 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     7501 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     7561 cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     7621 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     7681 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     7741 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     7801 ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     7861 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     7921 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
     7981 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
     8041 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
     8101 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     8161 gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     8221 cctgtacgag acacggatcg acctgtctca gctgggaggc gacaagcgtc ctgctgctac
     8281 taagaaagct ggtcaagcta agaaaaagaa agctagcggc agcggcgcca ccaacttcag
     8341 cctgctgaag caggccggcg acgtggagga gaaccccggc cccatgaccg agtacaagcc
     8401 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     8461 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     8521 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     8581 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     8641 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     8701 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     8761 caccgtcgga gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     8821 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     8881 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     8941 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc gcgatcgccg aaggatccgc
     9001 ggccgctgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc
     9061 ttccggaatg gagagcgacg agagcggcct gcccgccatg gagatcgagt gccgcatcac
     9121 cggcaccctg aacggcgtgg agttcgagct ggtgggcggc ggagagggca cccccaagca
     9181 gggccgcatg accaacaaga tgaaaagcac caaaggcgcc ctgaccttca gcccctacct
     9241 gctgagccac gtgatgggct acggcttcta ccacttcggc acctacccca gcggctacga
     9301 gaaccccttc ctgcacgcca tcaacaacgg cggctacacc aacacccgca tcgagaagta
     9361 cgaggacggc ggcgtgctgc acgtgagctt cagctaccgc tacgaggccg gccgcgtgat
     9421 cggcgacttc aaggtggtgg gcaccggctt ccccgaggac agcgtgatct tcaccgacaa
     9481 gatcatccgc agcaacgcca ccgtggagca cctgcacccc atgggcgata acgtgctggt
     9541 gggcagcttc gcccgcacct tcagcctgcg cgacggcggc tactacagct tcgtggtgga
     9601 cagccacatg cacttcaaga gcgccatcca ccccagcatc ctgcagaacg ggggccccat
     9661 gttcgccttc cgccgcgtgg aggagctgca cagcaacacc gagctgggca tcgtggagta
     9721 ccagcacgcc ttcaagaccc ccatcgcctt cgccagatcc cgcgctcagt cgtccaattc
     9781 tgccgtggac ggcaccgccg gacccggctc caccggatct cgctagagct gaatctaagt
     9841 cgacttaaga accgctcgag gccggcaagg ccggatccag acatgataag atacattgat
     9901 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     9961 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
    10021 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
    10081 aacctctaca aatgtggtat ggctgattat gatccggctg cctcgcgcgt ttcggtgatg
    10141 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
    10201 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
    10261 cagccatgag gtcgactcta gaggatcgat ccccgccccg gacgaactaa acctgactac
    10321 gacatctctg ccccttcttc gcggggcagt gcatgtaatc ccttcagttg gttggtacaa
    10381 cttgccaact gggccctgtt ccacatgtga cacggggggg gaccaaacac aaaggggttc
    10441 tctgactgta gttgacatcc ttataaatgg atgtgcacat ttgccaacac tgagtggctt
    10501 tcatcctgga gcagactttg cagtctgtgg actgcaacac aacattgcct ttatgtgtaa
    10561 ctcttggctg aagctcttac accaatgctg ggggacatgt acctcccagg ggcccaggaa
    10621 gactacggga ggctacacca acgtcaatca gaggggcctg tgtagctacc gataagcgga
    10681 ccctcaagag ggcattagca atagtgttta taaggccccc ttgttaaccc taaacgggta
    10741 gcatatgctt cccgggtagt agtatatact atccagacta accctaattc aatagcatat
    10801 gttacccaac gggaagcata tgctatcgaa ttagggttag taaaagggtc ctaaggaaca
    10861 gcgatatctc ccaccccatg agctgtcacg gttttattta catggggtca ggattccacg
    10921 agggtagtga accattttag tcacaagggc agtggctgaa gatcaaggag cgggcagtga
    10981 actctcctga atcttcgcct gcttcttcat tctccttcgt ttagctaata gaataactgc
    11041 tgagttgtga acagtaaggt gtatgtgagg tgctcgaaaa caaggtttca ggtgacgccc
    11101 ccagaataaa atttggacgg ggggttcagt ggtggcattg tgctatgaca ccaatataac
    11161 cctcacaaac cccttgggca ataaatacta gtgtaggaat gaaacattct gaatatcttt
    11221 aacaatagaa atccatgggg tggggacaag ccgtaaagac tggatgtcca tctcacacga
    11281 atttatggct atgggcaaca cataatccta gtgcaatatg atactggggt tattaagatg
    11341 tgtcccaggc agggaccaag acaggtgaac catgttgtta cactctattt gtaacaaggg
    11401 gaaagagagt ggacgccgac agcagcggac tccactggtt gtctctaaca cccccgaaaa
    11461 ttaaacgggg ctccacgcca atggggccca taaacaaaga caagtggcca ctcttttttt
    11521 tgaaattgtg gagtgggggc acgcgtcagc ccccacacgc cgccctgcgg ttttggactg
    11581 taaaataagg gtgtaataac ttggctgatt gtaaccccgc taaccactgc ggtcaaacca
    11641 cttgcccaca aaaccactaa tggcaccccg gggaatacct gcataagtag gtgggcgggc
    11701 caagataggg gcgcgattgc tgcgatctgg aggacaaatt acacacactt gcgcctgagc
    11761 gccaagcaca gggttgttgg tcctcatatt cacgaggtcg ctgagagcac ggtgggctaa
    11821 tgttgccatg ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta
    11881 tatctgggta gcataggcta tcctaatcta tatctgggta gtatatgcta tcctaatctg
    11941 tatccgggta gcatatgcta tcctaataga gattagggta gtatatgcta tcctaatcta
    12001 tatctgggta gcataggcta tcctaatcta tatctgggta gtatatgcta tcctaatctg
    12061 tatccgggta gcatatgcta tcctaataga gattagggta gtatatgcta tcctaattta
    12121 tatctgggta gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc
    12181 tgggtagcat atgctatcct aatctatatc tgggtagcat aggctatcct aatctatatc
    12241 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatttatatc
    12301 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
    12361 tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct catgcatata
    12421 cagtcagcat atgataccca gtagtagagt gggagtgcta tcctttgcat atgccgccac
    12481 ctcccaaggg ggcgtgaatt ttcgctgctt gtccttttcc tgctggttgc tcccattctt
    12541 aggtgaattt aaggaggcca ggctaaagcc gtcgcatgtc tgattgctca ccaggtaaat
    12601 gtcgctaatg ttttccaacg cgagaaggtg ttgagcgcgg agctgagtga cgtgacaaca
    12661 tgggtatgcc caattgcccc atgttgggag gacgaaaatg gtgacaagac agatggccag
    12721 aaatacacca acagcacgca tgatgtctac tggggattta ttctttagtg cgggggaata
    12781 cacggctttt aatacgattg agggcgtctc ctaacaagtt acatcactcc tgcccttcct
    12841 caccctcatc tccatcacct ccttcatctc cgtcatctcc gtcatcaccc tccgcggcag
    12901 ccccttccac cataggtgga aaccagggag gcaaatctac tccatcgtca aagctgcaca
    12961 cagtcaccct gatattgcag gtaggagcgg gctttgtcat aacaaggtcc ttaatcgcat
    13021 ccttcaaaac ctcagcaaat atatgagttt gtaaaaagac catgaaataa cagacaatgg
    13081 actcccttag cgggccaggt tgtgggccgg gtccaggggc cattccaaag gggagacgac
    13141 tcaatggtgt aagacgacat tgtggaatag caagggcagt tcctcgcctt aggttgtaaa
    13201 gggaggtctt actacctcca tatacgaaca caccggcgac ccaagttcct tcgtcggtag
    13261 tcctttctac gtgactccta gccaggagag ctcttaaacc ttctgcaatg ttctcaaatt
    13321 tcgggttgga acctccttga ccacgatgct ttccaaacca ccctcctttt ttgcgcctgc
    13381 ctccatcacc ctgaccccgg ggtccagtgc ttgggccttc tcctgggtca tctgcggggc
    13441 cctgctctat cgctcccggg ggcacgtcag gctcaccatc tgggccacct tcttggtggt
    13501 attcaaaata atcggcttcc cctacagggt ggaaaaatgg ccttctacct ggagggggcc
    13561 tgcgcggtgg agacccggat gatgatgact gactactggg actcctgggc ctcttttctc
    13621 cacgtccacg acctctcccc ctggctcttt cacgacttcc ccccctggct ctttcacgtc
    13681 ctctaccccg gcggcctcca ctacctcctc gaccccggcc tccactacct cctcgacccc
    13741 ggcctccact gcctcctcga ccccggcctc cacctcctgc tcctgcccct cctgctcctg
    13801 cccctcctcc tsctcctscc cctcctcccc ctcctsctcc tscccctcct ccccctcctc
    13861 ctcctscccc tcctscccct cctsctccts cccctcctsc ccctcctcct sctcctsccc
    13921 ctcctscccc tcctcctsct cctscccctc ctsctcctsc ycctccycct cctcccc
//
