LOCUS       sequence-337992-       10902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deletion of a genomic locus in Yap1 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10902)
  AUTHORS   Kagiwada S, Aramaki S, Wu G, Shin B, Kutejova E, Obridge D, Adachi 
            K, Wrana JL, Hubner K, Scholer HR
  TITLE     YAP establishes epiblast responsiveness to inductive signals for 
            germ cell fate.
  JOURNAL   Development. 2021 Oct 15;148(20). pii: 272520. doi: 
            10.1242/dev.199732. Epub 2021 Oct 19.
  PUBMED    34528691
REFERENCE   2  (bases 1 to 10902)
  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: 
            "Development."; date: "2021-10-15"; volume: "148(20). pii: 272520. 
            doi"; pages: " 10.1242/dev.199732. Epub 2021 Oct 19"
COMMENT     Sequence Label: pX330A-Yap1-E1-#2
FEATURES             Location/Qualifiers
     source          1..10902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          161..388
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      400..409
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             409..474
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             481..501
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             526..4626
                     /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             526..4626
                     /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             4627..4674
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4702..4719)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4708..4915
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4924..5064
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4924..5053
                     /label=AAV2 ITR
     promoter        5209..5420
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5433..5701
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      5730..5739
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5760..6422
                     /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"
     CDS             6492..6545
                     /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             6546..7145
                     /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(6546..6565)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7042..7062
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(7199..7219)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    7261..7382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7298..7317)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7352..7371
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7533..7988
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7620..7639)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7830..7851
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8005..8024)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8124..8146
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8184..8202)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8270..8374
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8375..8443,8444..9235)
                     /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(8593..8612)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9406..9994
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9895..9914
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        10056..10304
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     10056..10076
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     10227..10246
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        10326..10401
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        10498..10783
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggggcgcgcg ccrggggggg gggggggggg gggggggggg ggsggggsgr ggsggrgagg
       61 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
      121 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
      181 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
      241 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagctgag
      301 caagaggtaa gggtttaagg gatggttggt tggtggggta ttaatgttta attacctgga
      361 gcacctgcct gaaatcactt tttttcaggt tggaccggtg ccaccatgga ctataaggac
      421 cacgacggag actacaagga tcatgatatt gattacaaag acgatgacga taagatggcc
      481 ccaaagaaga agcggaaggt cggtatccac ggagtcccag cagccgacaa gaagtacagc
      541 atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag
      601 gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac
      661 ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga
      721 accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc
      781 agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg
      841 gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg
      901 gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc
      961 gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc
     1021 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     1081 cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg
     1141 gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc
     1201 gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg
     1261 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg
     1321 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     1381 gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg
     1441 agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac
     1501 gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag
     1561 tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga
     1621 gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc
     1681 gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac
     1741 aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag
     1801 gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc
     1861 cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc
     1921 agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct
     1981 tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag
     2041 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa
     2101 gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag
     2161 gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag
     2221 gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg
     2281 ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc
     2341 ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt
     2401 gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa
     2461 gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg
     2521 atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac
     2581 ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag
     2641 gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat
     2701 ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag
     2761 ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag
     2821 aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag
     2881 ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg
     2941 cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag
     3001 gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt
     3061 ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag
     3121 agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg
     3181 ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc
     3241 ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag
     3301 atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat
     3361 gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc
     3421 cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac
     3481 gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc
     3541 gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag
     3601 caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc
     3661 aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac
     3721 ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg
     3781 ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc
     3841 aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg
     3901 gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg
     3961 gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc
     4021 accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc
     4081 tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg
     4141 gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg
     4201 gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag
     4261 ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg
     4321 gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat
     4381 ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc
     4441 gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac
     4501 tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc
     4561 ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga
     4621 ggcgacaaaa ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaagtaagaa
     4681 ttcctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     4741 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     4801 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     4861 tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct ggggagcggc
     4921 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4981 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     5041 agcgagcgcg cagctgcctg caggaaagag cccacaaccc ctcacttggt gggccagtcc
     5101 tctgatagac tgtgtcccct ggatacccgt attctagagc tagcgaattc gaatttaaat
     5161 cggatccgcg gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca
     5221 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga
     5281 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     5341 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     5401 ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc
     5461 gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct
     5521 gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg
     5581 gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc
     5641 tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat
     5701 ccaagctgtg accggcgcct acgctagacg ccaccatgga gagcgacgag agcggcctgc
     5761 ccgccatgga gatcgagtgc cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg
     5821 tgggcggcgg agagggcacc cccaagcagg gccgcatgac caacaagatg aagagcacca
     5881 aaggcgccct gaccttcagc ccctacctgc tgagccacgt gatgggctac ggcttctacc
     5941 acttcggcac ctaccccagc ggctacgaga accccttcct gcacgccatc aacaacggcg
     6001 gctacaccaa cacccgcatc gagaagtacg aggacggcgg cgtgctgcac gtgagcttca
     6061 gctaccgcta cgaggccggc cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc
     6121 ccgaggacag cgtgatcttc accgacaaga tcatccgcag caacgccacc gtggagcacc
     6181 tgcaccccat gggcgataac gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg
     6241 acggcggcta ctacagcttc gtggtggaca gccacatgca cttcaagagc gccatccacc
     6301 ccagcatcct gcagaacggg ggccccatgt tcgccttccg ccgcgtggag gagctgcaca
     6361 gcaacaccga gctgggcatc gtggagtacc agcacgcctt caagaccccc atcgccttcg
     6421 ccagatcccg cgctcagtcg tccaattctg ccgtggacgg caccgccgga cccggctcca
     6481 ccggatctcg cgagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg
     6541 gccctatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     6601 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     6661 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     6721 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     6781 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     6841 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     6901 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     6961 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     7021 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     7081 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     7141 cctgaaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     7201 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gcgttaacta
     7261 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     7321 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     7381 tatcatgtct ggaattgact caaatgatgt caattagtct atcagaagct catctggtct
     7441 cccttccggg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     7501 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     7561 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     7621 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     7681 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     7741 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     7801 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     7861 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     7921 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     7981 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     8041 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     8101 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     8161 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     8221 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     8281 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     8341 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     8401 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     8461 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     8521 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     8581 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     8641 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     8701 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     8761 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     8821 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     8881 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     8941 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     9001 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     9061 tgctgataaa tctggagccg gtgagcgtgg aagccgcggt atcattgcag cactggggcc
     9121 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     9181 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     9241 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     9301 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     9361 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     9421 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     9481 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     9541 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     9601 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     9661 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     9721 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     9781 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     9841 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     9901 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     9961 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
    10021 gttcctggcc ttttgctggc cttttgctca catgtgaggg cctatttccc atgattcctt
    10081 catatttgca tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa
    10141 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
    10201 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
    10261 tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgccccc gggtcacgtg
    10321 cggccgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
    10381 aaagtggcac cgagtcggtg cttttttgtt ttagagctag aaatagcaag ttaaaataag
    10441 gctagtccgt ttttagcgcg tgcgccaatt ctgcagacaa atggctctag aggtacccgt
    10501 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
    10561 gtcaatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
    10621 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
    10681 caatgacggt aaatggcccg cctggcattg tgcccagtac atgaccttat gggactttcc
    10741 tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac
    10801 gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat
    10861 tttttaatta ttttgtgcag cgatgggggc gggggggggg gg
//
