LOCUS       sequence-353288-        9127 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 9127)
  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 9127)
  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-exon1
FEATURES             Location/Qualifiers
     source          1..9127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          117..344
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      356..365
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             365..430
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             437..457
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             482..4582
                     /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             482..4582
                     /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             4583..4630
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4646..4699
                     /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             4700..5299
                     /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(4700..4719)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(5327..5344)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5333..5540
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5549..5689
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5549..5678
                     /label=AAV2 ITR
     rep_origin      5764..6219
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5851..5870)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6061..6082
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6236..6255)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6355..6377
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6415..6433)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6501..6605
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6606..6674,6675..7466)
                     /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(6824..6843)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7637..8225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8126..8145
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8287..8535
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8287..8307
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8458..8477
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8557..8632
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8729..9014
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
       61 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      121 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      181 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      241 gctgtaatta gctgagcaag aggtaagggt ttaagggatg gttggttggt ggggtattaa
      301 tgtttaatta cctggagcac ctgcctgaaa tcactttttt tcaggttgga ccggtgccac
      361 catggactat aaggaccacg acggagacta caaggatcat gatattgatt acaaagacga
      421 tgacgataag atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc
      481 cgacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct gggccgtgat
      541 caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca
      601 cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc
      661 cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta
      721 tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct tccacagact
      781 ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa
      841 catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa
      901 actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat
      961 gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt
     1021 ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat
     1081 caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg
     1141 gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct
     1201 gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga
     1261 tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca
     1321 gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct
     1381 gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat
     1441 gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca
     1501 gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg
     1561 ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga
     1621 aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa
     1681 gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc
     1741 cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga
     1801 gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag
     1861 attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt
     1921 ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa
     1981 cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta
     2041 taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag
     2101 cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt
     2161 gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc
     2221 cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat
     2281 caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct
     2341 gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca
     2401 cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag
     2461 gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga
     2521 tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag
     2581 cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca
     2641 cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt
     2701 gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat
     2761 cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat
     2821 gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt
     2881 ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga
     2941 tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat
     3001 cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga
     3061 caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa
     3121 ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac
     3181 caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct
     3241 ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac
     3301 taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa
     3361 gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta
     3421 ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta
     3481 ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat
     3541 gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa
     3601 catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc
     3661 tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc
     3721 caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca
     3781 gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc
     3841 cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta
     3901 ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa
     3961 agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt
     4021 tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta
     4081 ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca
     4141 gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca
     4201 ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca
     4261 gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat
     4321 cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc
     4381 catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc
     4441 tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga
     4501 ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga
     4561 cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa
     4621 aaagaaaaag gaattcggca gtggagaggg cagaggaagt ctgctaacat gcggtgacgt
     4681 cgaggagaat cctggcccaa tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     4741 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     4801 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     4861 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     4921 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     4981 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     5041 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     5101 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     5161 cggggtgccc gccttcctgg agacctccgc gccccacaac ctccccttct acgagcggct
     5221 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     5281 ccgcaagccc ggtgcctgag aattctaact agagctcgct gatcagcctc gactgtgcct
     5341 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     5401 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     5461 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagag
     5521 aatagcaggc atgctgggga gcggccgcag gaacccctag tgatggagtt ggccactccc
     5581 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     5641 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     5701 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     5761 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     5821 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     5881 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5941 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     6001 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6061 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     6121 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     6181 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     6241 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     6301 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     6361 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     6421 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     6481 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6541 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6601 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6661 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6721 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6781 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6841 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6901 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6961 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     7021 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7081 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7141 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     7201 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     7261 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     7321 gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     7381 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     7441 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7501 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7561 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7621 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7681 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7741 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     7801 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7861 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7921 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7981 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     8041 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     8101 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     8161 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     8221 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     8281 acatgtgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     8341 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     8401 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     8461 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     8521 aaaggacgaa acaccggtct ctcagacccg gagaacgttt tagagctaga aatagcaagt
     8581 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttgt
     8641 tttagagcta gaaatagcaa gttaaaataa ggctagtccg tttttagcgc gtgcgccaat
     8701 tctgcagaca aatggctcta gaggtacccg ttacataact tacggtaaat ggcccgcctg
     8761 gctgaccgcc caacgacccc cgcccattga cgtcaatagt aacgccaata gggactttcc
     8821 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     8881 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     8941 gtgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     9001 tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc
     9061 cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg
     9121 cgggggg
//
