LOCUS       sequence-340334-        9143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A knock-out vector for the dog Egf.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9143)
  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 9143)
  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
FEATURES             Location/Qualifiers
     source          1..9143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     misc_RNA        complement(501..576)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(598..846)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(656..675)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(826..846)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(908..1496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(988..1007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1667..2458,2459..2527))
                     /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     2290..2309
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2528..2632)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2700..2718
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2756..2778)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2878..2897
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2914..3369)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3051..3072)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3263..3282
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3444..3584
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3455..3584
                     /label=AAV2 ITR
     polyA_signal    3593..3800
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3789..3806
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3834..4433)
                     /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     4414..4433
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(4434..4487)
                     /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             complement(4503..4550)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(4551..8651)
                     /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             complement(4551..8651)
                     /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             complement(8676..8696)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8703..8768)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      8768..8777
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          8789..9016
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
ORIGIN
        1 cccccccccc cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg
       61 ggggtgggga ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca
      121 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
      181 tgtactgggc acaatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
      241 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
      301 attgacgtca atggaaagtc cctattggcg ttactattga cgtcaatggg cgggggtcgt
      361 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgggtacc tctagagcca
      421 tttgtctgca gaattggcgc acgcgctaaa aacggactag ccttatttta acttgctatt
      481 tctagctcta aaacaaaaaa gcaccgactc ggtgccactt tttcaagttg ataacggact
      541 agccttattt taacttgcta tttctagctc taaaacctta tcattctgtg gccggtcggt
      601 gtttcgtcct ttccacaaga tatataaagc caagaaatcg aaatactttc aagttacggt
      661 aagcatatga tagtccattt taaaacataa ttttaaaact gcaaactacc caagaaatta
      721 ttactttcta cgtcacgtat tttgtactaa tatctttgtg tttacagtca aattaattcc
      781 aattatctct ctaacagcct tgtatcgtat atgcaaatat gaaggaatca tgggaaatag
      841 gccctcacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      901 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      961 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1021 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1081 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1141 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1201 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1261 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1321 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1381 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1441 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1501 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1561 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1621 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     1681 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     1741 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     1801 taccgcggct tccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     1861 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     1921 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     1981 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     2041 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     2101 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     2161 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     2221 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     2281 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     2341 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     2401 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     2461 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     2521 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     2581 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     2641 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     2701 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     2761 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     2821 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     2881 catcagagca gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa
     2941 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaatagacc gaaatcggca
     3001 aaatccctta taaatcaaaa gaatagcccg agatagagtt gagtgttgtt ccagtttgga
     3061 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3121 agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc
     3181 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3241 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg
     3301 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3361 agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag
     3421 gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc
     3481 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
     3541 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc gctccccagc
     3601 atgcctgcta ttctcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag
     3661 aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac
     3721 agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg
     3781 ctggcaacta gaaggcacag tcgaggctga tcagcgagct ctagttagaa ttctcaggca
     3841 ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg
     3901 acggtgaagc cgagccgctc gtagaagggg aggttgtggg gcgcggaggt ctccaggaag
     3961 gcgggcaccc cggcgcgctc ggccgcctcc actccgggga gcacgacggc gctgcccaga
     4021 cccttgccct ggtggtcggg cgagacgccg acggtggcca ggaaccacgc gggctccttg
     4081 ggccggtgcg gcgccaggag gccttccatc tgttgctgcg cggccagccg ggaaccgctc
     4141 aactcggcca tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac gctctccggc
     4201 gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc
     4261 ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg gcggtccgga
     4321 tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg cggcgagggt gcgtacggcc
     4381 ctggggacgt cgtcgcgggt ggcgaggcgc accgtgggct tgtactcggt cattgggcca
     4441 ggattctcct cgacgtcacc gcatgttagc agacttcctc tgccctctcc actgccgaat
     4501 tcctttttct tttttgcctg gccggccttt ttcgtggccg ccggcctttt gtcgcctccc
     4561 agctgagaca ggtcgatccg tgtctcgtac aggccggtga tgctctggtg gatcagggtg
     4621 gcgtccagca cctctttggt gctggtgtac ctcttccggt cgatggtggt gtcaaagtac
     4681 ttgaaggcgg caggggctcc cagattggtc agggtaaaca ggtggatgat attctcggcc
     4741 tgctctctga tgggcttatc ccggtgcttg ttgtaggcgg acagcacttt gtccagatta
     4801 gcgtcggcca ggatcactct cttggagaac tcgctgatct gctcgatgat ctcgtccagg
     4861 tagtgcttgt gctgttccac aaacagctgt ttctgctcat tatcctcggg ggagcccttc
     4921 agcttctcat agtggctggc caggtacagg aagttcacat atttggaggg cagggccagt
     4981 tcgtttccct tctgcagttc gccggcagag gccagcattc tcttccggcc gttttccagc
     5041 tcgaacaggg agtacttagg cagcttgatg atcaggtcct ttttcacttc tttgtagccc
     5101 ttggcttcca gaaagtcgat gggattcttc tcgaagctgc ttctttccat gatggtgatc
     5161 cccagcagct ctttcacact cttcagtttc ttggacttgc ccttttccac tttggccacc
     5221 accagcacag aataggccac ggtggggctg tcgaagccgc cgtacttctt agggtcccag
     5281 tccttctttc tggcgatcag cttatcgctg ttcctcttgg gcaggataga ctctttgctg
     5341 aagccgcctg tctgcacctc ggtctttttc acgatattca cttggggcat gctcagcact
     5401 ttccgcacgg tggcaaaatc ccggccctta tcccacacga tctccccggt ttcgccgttt
     5461 gtctcgatca gaggccgctt ccggatctcg ccgttggcca gggtaatctc ggtcttgaaa
     5521 aagttcatga tgttgctgta gaagaagtac ttggcggtag ccttgccgat ttcctgctcg
     5581 ctcttggcga tcatcttccg cacgtcgtac accttgtagt cgccgtacac gaactcgctt
     5641 tccagcttag ggtacttttt gatcagggcg gttcccacga cggcgttcag gtaggcgtcg
     5701 tgggcgtggt ggtagttgtt gatctcgcgc actttgtaaa actggaaatc cttccggaaa
     5761 tcggacacca gcttggactt cagggtgatc actttcactt cccggatcag cttgtcattc
     5821 tcgtcgtact tagtgttcat ccgggagtcc aggatctgtg ccacgtgctt tgtgatctgc
     5881 cgggtttcca ccagctgtct cttgatgaag ccggccttat ccagttcgct caggccgcct
     5941 ctctcggcct tggtcagatt gtcgaacttt ctctgggtaa tcagcttggc gttcagcagc
     6001 tgccgccagt agttcttcat cttcttcacg acctcttcgg agggcacgtt gtcgctcttg
     6061 ccccggttct tgtcgcttct ggtcagcacc ttgttgtcga tggagtcgtc cttcagaaag
     6121 ctctgaggca cgatatggtc cacatcgtag tcggacagcc ggttgatgtc cagttcctgg
     6181 tccacgtaca tatcccgccc attctgcagg tagtacaggt acagcttctc gttctgcagc
     6241 tgggtgtttt ccacggggtg ttctttcagg atctggctgc ccagctcttt gatgccctct
     6301 tcgatccgct tcattctctc gcggctgttc ttctgtccct tctgggtggt ctggttctct
     6361 ctggccattt cgatcacgat gttctcgggc ttgtgccggc ccatcacttt cacgagctcg
     6421 tccaccacct tcactgtctg caggatgccc ttcttaatgg cggggctgcc ggccagattg
     6481 gcaatgtgct cgtgcaggct atcgccctgg ccggacacct gggctttctg gatgtcctct
     6541 ttaaaggtca ggctgtcgtc gtggatcagc tgcatgaagt ttctgttggc gaagccgtcg
     6601 gacttcagga aatccaggat tgtcttgccg gactgcttgt cccggatgcc gttgatcagc
     6661 ttccggctca gcctgcccca gccggtgtat ctccgccgct tcagctgctt catcactttg
     6721 tcgtcgaaca ggtgggcata ggttttcagc cgttcctcga tcatctctct gtcctcaaac
     6781 agtgtcaggg tcagcacgat atcttccaga atgtcctcgt tttcctcatt gtccaggaag
     6841 tccttgtcct tgataatttt cagcagatcg tggtatgtgc ccagggaggc gttgaaccga
     6901 tcttccacgc cggagatttc cacggagtcg aagcactcga ttttcttgaa gtagtcctct
     6961 ttcagctgct tcacggtcac tttccggttg gtcttgaaca gcaggtccac gatggccttt
     7021 ttctgctcgc cgctcaggaa ggcgggcttt ctcattccct cggtcacgta tttcactttg
     7081 gtcagctcgt tatacacggt gaagtactcg tacagcaggc tgtgcttggg cagcaccttc
     7141 tcgttgggca ggttcttatc gaagttggtc atccgctcga tgaagctctg ggcggaagcg
     7201 cccttgtcca ccacttcctc gaagttccag ggggtgatgg tttcctcgct ctttctggtc
     7261 atccaggcga atctgctgtt tcccctggcc agagggccca cgtagtaggg gatgcggaag
     7321 gtcaggatct tctcgatctt ttcccggttg tccttcagga atgggtaaaa atcttcctgc
     7381 cgccgcagaa tggcgtgcag ctctcccagg tggatctggt gggggatgct gccgttgtcg
     7441 aaggtccgct gcttccgcag caggtcctct ctgttcagct tcacgagcag ttcctcggtg
     7501 ccgtccatct tttccaggat gggcttgatg aacttgtaga actcttcctg gctggctccg
     7561 ccgtcaatgt agccggcgta gccgttcttg ctctggtcga agaaaatctc tttgtacttc
     7621 tcaggcagct gctgccgcac gagagctttc agcagggtca ggtcctggtg gtgctcgtcg
     7681 tatctcttga tcatagaggc gctcaggggg gccttggtga tctcggtgtt cactctcagg
     7741 atgtcgctca gcaggatggc gtcggacagg ttcttggcgg ccagaaacag gtcggcgtac
     7801 tggtcgccga tctgggccag caggttgtcc aggtcgtcgt cgtaggtgtc cttgctcagc
     7861 tgcagtttgg catcctcggc caggtcgaag ttgctcttga agttgggggt caggcccagg
     7921 ctcagggcaa tcaggtttcc gaacaggcca ttcttcttct cgccgggcag ctgggcgatc
     7981 agattttcca gccgtctgct cttgctcagt ctggcagaca ggatggcctt ggcgtccacg
     8041 ccgctggcgt tgatggggtt ttcctcgaac agctggttgt aggtctgcac cagctggatg
     8101 aacagcttgt ccacgtcgct gttgtcgggg ttcaggtcgc cctcgatcag gaagtggccc
     8161 cggaacttga tcatgtgggc cagggccaga tagatcagcc gcaggtcggc cttgtcggtg
     8221 ctgtccacca gtttctttct caggtggtag atggtggggt acttctcgtg gtaggccacc
     8281 tcgtccacga tgttgccgaa gatggggtgc cgctcgtgct tcttatcctc ttccaccagg
     8341 aaggactctt ccagtctgtg gaagaagctg tcgtccacct tggccatctc gttgctgaag
     8401 atctcttgca gatagcagat ccggttcttc cgtctggtgt atcttcttct ggcggttctc
     8461 ttcagccggg tggcctcggc tgtttcgccg ctgtcgaaca gcagggctcc gatcaggttc
     8521 ttcttgatgc tgtgccggtc ggtgttgccc agcaccttga atttcttgct gggcaccttg
     8581 tactcgtcgg tgatcacggc ccagcccaca gagttggtgc cgatgtccag gccgatgctg
     8641 tacttcttgt cggctgctgg gactccgtgg ataccgacct tccgcttctt ctttggggcc
     8701 atcttatcgt catcgtcttt gtaatcaata tcatgatcct tgtagtctcc gtcgtggtcc
     8761 ttatagtcca tggtggcacc ggtccaacct gaaaaaaagt gatttcaggc aggtgctcca
     8821 ggtaattaaa cattaatacc ccaccaacca accatccctt aaacccttac ctcttgctca
     8881 gctaattaca gcccggagga gaagggccgt cccgcccgct cacctgtggg agtaacgcgg
     8941 tcagtcagag ccggggcggg cggcgcgagg cggcggcgga gcggggcacg gggcgaaggc
     9001 agcgcgcagc gactcccgcc cgccgcgcgc ttcgcttttt atagggccgc cgccgccgcc
     9061 gcctcgccat aaaaggaaac tttcggagcg cgccgctctg attggctgcc gccgcacctc
     9121 tccgcctcgc cccgccccgc ccc
//
