LOCUS       sequence-353287-        9120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A knockout vector for the dog Egf.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9120)
  AUTHORS   Lin S, Hirayama D, Maryu G, Matsuda K, Hino N, Deguchi E, Aoki K, 
            Iwamoto R, Terai K, Matsuda M
  TITLE     Redundant roles of EGFR ligands in the ERK activation waves during 
            collective cell migration.
  JOURNAL   Life Sci Alliance. 2021 Oct 19;5(1). pii: 5/1/e202101206. doi: 
            10.26508/lsa.202101206. Print 2022 Jan.
  PUBMED    34667080
REFERENCE   2  (bases 1 to 9120)
  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: "Life Sci 
            Alliance."; date: "2021-10-19"; volume: "5(1). pii: 5/1/e202101206. 
            doi"; pages: " 10.26508/lsa.202101206. Print 2022 Jan"
COMMENT     Sequence Label: pX459-dEGF-5'UTR
FEATURES             Location/Qualifiers
     source          1..9120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          110..337
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      349..358
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             358..423
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             430..450
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             475..4575
                     /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             475..4575
                     /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             4576..4623
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4639..4692
                     /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             4693..5292
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4693..4712)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(5320..5337)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5326..5533
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5542..5682
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5542..5671
                     /label=AAV2 ITR
     rep_origin      5757..6212
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5844..5863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6054..6075
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6229..6248)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6348..6370
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6408..6426)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6494..6598
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6599..6667,6668..7459)
                     /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(6817..6836)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7630..8218
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8119..8138
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8280..8528
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8280..8300
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8451..8470
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8550..8625
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8722..9007
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
       61 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
      121 gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg
      181 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa
      241 ttagctgagc aagaggtaag ggtttaaggg atggttggtt ggtggggtat taatgtttaa
      301 ttacctggag cacctgcctg aaatcacttt ttttcaggtt ggaccggtgc caccatggac
      361 tataaggacc acgacggaga ctacaaggat catgatattg attacaaaga cgatgacgat
      421 aagatggccc caaagaagaa gcggaaggtc ggtatccacg gagtcccagc agccgacaag
      481 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac
      541 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
      601 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
      661 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
      721 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
      781 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
      841 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
      901 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
      961 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     1021 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     1081 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     1141 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc
     1201 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     1261 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     1321 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     1381 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     1441 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     1501 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
     1561 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     1621 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     1681 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     1741 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     1801 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     1861 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     1921 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     1981 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
     2041 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     2101 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     2161 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     2221 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     2281 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     2341 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     2401 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     2461 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     2521 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
     2581 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     2641 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     2701 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
     2761 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
     2821 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
     2881 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
     2941 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct
     3001 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac
     3061 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
     3121 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
     3181 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
     3241 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
     3301 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
     3361 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
     3421 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
     3481 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
     3541 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
     3601 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
     3661 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
     3721 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
     3781 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
     3841 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
     3901 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
     3961 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
     4021 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
     4081 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
     4141 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
     4201 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
     4261 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
     4321 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
     4381 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
     4441 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
     4501 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
     4561 cagctgggag gcgacaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa
     4621 aaggaattcg gcagtggaga gggcagagga agtctgctaa catgcggtga cgtcgaggag
     4681 aatcctggcc caatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     4741 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     4801 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     4861 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     4921 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     4981 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     5041 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     5101 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     5161 cccgccttcc tggagacctc cgcgccccac aacctcccct tctacgagcg gctcggcttc
     5221 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     5281 cccggtgcct gagaattcta actagagctc gctgatcagc ctcgactgtg ccttctagtt
     5341 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     5401 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     5461 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gagaatagca
     5521 ggcatgctgg ggagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     5581 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     5641 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     5701 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     5761 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5821 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5881 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5941 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     6001 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     6061 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     6121 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     6181 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     6241 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     6301 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     6361 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     6421 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     6481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     7021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     7081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     7141 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     7201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     7261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg
     7321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     7381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     7441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     7501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7741 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     8041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     8101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     8161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     8221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtg
     8281 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
     8341 taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
     8401 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
     8461 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
     8521 gaaacaccgc aaaaagagac ccggtgtttg ttttagagct agaaatagca agttaaaata
     8581 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag
     8641 ctagaaatag caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag
     8701 acaaatggct ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc
     8761 gcccaacgac ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg
     8821 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     8881 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca
     8941 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     9001 taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
     9061 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
//
