LOCUS       sequence-328056-       13338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAVS1 targeting vector carrying CAG-SpCas9-neo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13338)
  TITLE     Cas9 expression in iPSCs using AAVS1-CAGCas9neo
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13338)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAVS1-CAG-hCAS9-neo
FEATURES             Location/Qualifiers
     source          1..13338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..502
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    579..603
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     enhancer        625..1004
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1282..2298
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2222..2244
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2306..2325
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2366..2431
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2438..2458
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2483..6583
                     /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             2483..6583
                     /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             6584..6631
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     misc_feature    join(6660..7234,7235..7237,7238..7246)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6827..6844)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7054..7073
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7247..8050
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7310..7329)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7920..7939
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8082..8099)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8088..8312
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(8357..8381)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    8554..9083
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(9583..9600)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9583..9599)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9609..9732
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             10232..10423
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(10417..10439)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    10525..10665
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     10580..10599
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(10680..10697)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10851..11439)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10931..10950)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11610..12401,12402..12470))
                     /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     12233..12252
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12471..12575)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     12643..12661
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    complement(12687..12810)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gagcacttcc ttctcggcgc tgcaccacgt gatgtcctct gagcggatcc tccccgtgtc
       61 tgggtcctct ccgggcatct ctcctccctc acccaacccc atgccgtctt cactcgctgg
      121 gttccctttt ccttctcctt ctggggcctg tgccatctct cgtttcttag gatggccttc
      181 tccgacggat gtctcccttg cgtcccgcct ccccttcttg taggcctgca tcatcaccgt
      241 ttttctggac aaccccaaag taccccgtct ccctggcttt agccacctct ccatcctctt
      301 gctttctttg cctggacacc ccgttctcct gtggattcgg gtcacctctc actcctttca
      361 tttgggcagc tcccctaccc cccttacctc tctagtctgt gctagctctt ccagccccct
      421 gtcatggcat cttccagggg tccgagagct cagctagtct tcttcctcca acccgggccc
      481 ctatgtccac ttcaggacag catgtttgct gcctccaggg atcctgtgtc cccgagctgg
      541 gaccacctta taaggcgcat aacgatacca cgatatcaac aagtttgtac aaaaaagcag
      601 gctggcgccg gaaccaattc agtcgacatt gattattgac tagttattaa tagtaatcaa
      661 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      721 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      781 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      841 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      901 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      961 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
     1021 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
     1081 ttttttaatt attttgtgca gcgatggggg cggggggggg ggnnnnnnnc gccaggcggg
     1141 gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag
     1201 agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa
     1261 aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc
     1321 gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc
     1381 gggcgggacg gcccttctcc tccgggctgt aattagcgct tggtttaatg acggcttgtt
     1441 tcttttctgt ggctgcgtga aagccttgag gggctccggg agggcccttt gtgcgggggg
     1501 agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg
     1561 ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg
     1621 cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac
     1681 aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc
     1741 gggctgcaac cccccctgca cccccctccc cgagttgctg agcacggccc ggcttcgggt
     1801 gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag
     1861 gtgggggtgc cgggcggggc ggggccgcct cgggccgggg agggctcggg ggaggggcgc
     1921 ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca ttgcctttta
     1981 tggtaatcgt gcgagagggc gcagggactt cctttgtccc aaatctgtgc ggagccgaaa
     2041 tctgggaggc gccgccgcac cccctctagc gggcgcgggg cgaagcggtg cggcgccggc
     2101 aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct
     2161 ctccagcctc ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg
     2221 gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc
     2281 ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt
     2341 ggcaaagaat tcgtcgacca ccatggacta taaggaccac gacggagact acaaggatca
     2401 tgatattgat tacaaagacg atgacgataa gatggcccca aagaagaagc ggaaggtcgg
     2461 tatccacgga gtcccagcag ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa
     2521 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
     2581 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
     2641 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
     2701 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     2761 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     2821 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     2881 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     2941 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     3001 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     3061 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     3121 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     3181 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     3241 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     3301 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     3361 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     3421 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     3481 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     3541 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     3601 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     3661 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     3721 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     3781 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     3841 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     3901 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     3961 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     4021 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     4081 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     4141 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     4201 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     4261 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     4321 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     4381 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     4441 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     4501 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     4561 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     4621 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     4681 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     4741 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     4801 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     4861 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     4921 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     4981 cgactacgat gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa
     5041 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     5101 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     5161 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     5221 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     5281 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     5341 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     5401 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     5461 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     5521 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     5581 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     5641 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     5701 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     5761 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     5821 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     5881 cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     5941 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     6001 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     6061 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     6121 ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     6181 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     6241 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
     6301 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
     6361 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
     6421 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     6481 gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     6541 cctgtacgag acacggatcg acctgtctca gctgggaggc gacaaaaggc cggcggccac
     6601 gaaaaaggcc ggccaggcaa aaaagaaaaa gtaagaattc tagcggccgc cgcccccccc
     6661 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     6721 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     6781 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     6841 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     6901 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     6961 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     7021 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     7081 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
     7141 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     7201 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg gatcggccat
     7261 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     7321 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     7381 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     7441 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     7501 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     7561 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     7621 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     7681 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     7741 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     7801 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     7861 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     7921 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     7981 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     8041 gttcttctga cgtacgcgat gatctagagc tcgctgatca gcctcgactg tgccttctag
     8101 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     8161 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     8221 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     8281 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa cctgcaacta
     8341 gtctcgagat atctagaccc agctttcttg tacaaagtgg ttgatatctc tatagtcgca
     8401 gtaggcggtc cttaggcctc ctccttccta gtctcctgat attgggtcta acccccacct
     8461 cctgttaggc agattcctta tctggtgaca cacccccatt tcctggagcc atctctctcc
     8521 ttgccagaac ctctaaggtt tgcttacgat ggagccagag aggatcctgg gagggagagc
     8581 ttggcagggg gtgggaggga agggggggat gcgtgacctg cccggttctc agtggccacc
     8641 ctgcgctacc ctctcccaga acctgagctg ctctgacgcg gctgtctggt gcgtttcact
     8701 gatcctggtg ctgcagcttc cttacacttc ccaagaggag aagcagtttg gaaaaacaaa
     8761 atcagaataa gttggtcctg agttctaact ttggctcttc acctttctag tccccaattt
     8821 atattgttcc tccgtgcgtc agttttacct gtgagataag gccagtagcc agccccgtcc
     8881 tggcagggct gtggtgagga ggggggtgtc cgtgtggaaa actccctttg tgagaatggt
     8941 gcgtcctagg tgttcaccag gtcgtggccg cctctactcc ctttctcttt ctccatcctt
     9001 ctttccttaa agagtcccca gtgctatctg ggacatattc ctccgcccag agcagggtcc
     9061 cgcttcccta aggccctgct ctgggcttct gggtttgagt ccttggcaag cccaggagag
     9121 gcgctcaggc ttccctgtcc cccttcctcg tccaccatct catgcccctg gctctcctgc
     9181 cccttcccta caggggttcc tggctctgct cttcagactg agccccgttc ccctgcatcc
     9241 ccgttcccct gcatccccct tcccctgcat cccccagagg ccccaggcca cctacttggc
     9301 ctggacccca cgagaggcca ccccagccct gtctaccagg ctgccttttg ggtggattct
     9361 cctccaactg tggggtgact gcttggcaaa ctcactcttc ggggtatccc aggaggcctg
     9421 gagcattggg gtgggctggg gttcagagag gagggattcc cttctcaggt tacgtggcca
     9481 agaagcaggg gagctgggtt tgggtcaggt ctgggtgtgg ggtgaccagc ttatgctgtt
     9541 tgcccaggac agcctagttt tagcactgaa accctcagtc ccactggccg tcgttttaca
     9601 ttaattaaca actttgtata gaaaagttga acgagaaacg taaaatgata taaatatcaa
     9661 tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat
     9721 ccagtcacta tggcggccgc aatcgggcag cgttgggtcc tggccacggg tgcgcatgat
     9781 cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa
     9841 tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg
     9901 agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc
     9961 agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg
    10021 aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc
    10081 ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc
    10141 atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat
    10201 gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa
    10261 catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga
    10321 cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg
    10381 cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
    10441 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
    10501 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
    10561 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
    10621 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
    10681 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
    10741 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
    10801 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
    10861 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
    10921 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
    10981 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
    11041 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
    11101 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
    11161 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
    11221 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
    11281 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
    11341 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
    11401 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
    11461 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
    11521 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
    11581 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
    11641 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
    11701 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
    11761 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
    11821 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
    11881 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg
    11941 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
    12001 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
    12061 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
    12121 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
    12181 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc
    12241 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
    12301 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
    12361 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    12421 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
    12481 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
    12541 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
    12601 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
    12661 ccctttcgtc ttcaagaatt gtcgaccata gtgactggat atgttgtgtt ttacagtatt
    12721 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
    12781 tttctcgttc aactttatta tacaaagttg cggacgcgat cgccggcgga taacaatttc
    12841 acacaggatt accatccctc cctcgacttc ccctcttccg atgttgagcc cctccagccg
    12901 gtcctggact ttgtctcctt ccctgccctg ccctctcctg aacctgagcc agctcccata
    12961 gctcagtctg gtctatctgc ctggccctgg ccattgtcac tttgcgctgc cctcctctcg
    13021 cccccgagtg cccttgctgt gccgccggaa ctctgccctc taacgctgcc gtctctctcc
    13081 tgagtccgga ccactttgag ctctactggc ttctgcgccg cctctggccc actgtttccc
    13141 cttcccaggc aggtcctgct ttctctgacc tgcattctct cccctgggcc tgtgccgctt
    13201 tctgtctgca gcttgtggcc tgggtcacct ctacggctgg cccagatcct tccctgccgc
    13261 ctccttcagg ttccgtcttc ctccactccc tcttcccctt gctctctgct gtgttgctgc
    13321 ccaaggatgc tctttccg
//
