LOCUS       sequence-353286-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A knockout vector for the dog Adam17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  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 13013)
  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: pLentiCRISPRv2-dADAM17 (#1)
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..30)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(184..772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(264..283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(943..1734,1735..1803))
                     /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     1566..1585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1804..1908)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1983..2002)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2174..2553
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2555..2753
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2708..2728
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2771..2951
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2998..3123
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3616..3849
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4034..4078
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    4376..4493
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4544..4792
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4544..4564
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4715..4734
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4814..4889
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4951..5162
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5181..5190
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5187..9290
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5187..9290
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             9291..9338
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9339..9362
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9372..9428
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             9429..10025
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     9922..9942
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10041..10629
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10094..10114)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10512..10523)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10701..10934
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10960..10977)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10966..11190
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11236..11664
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11323..11342)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11533..11554
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11673..11693)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11678..12007
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11858..11993
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11920..11939
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12055..12102
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12121..12495
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    12625..12746
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12662..12681)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12716..12735
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12795..12811)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12795..12811)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12808..12830)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12819..12835
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(12843..12849,12850..12867,12868..12873))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12888..12909
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
       61 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      121 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      181 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      241 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      301 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      361 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      421 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      481 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      541 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      601 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      661 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      721 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      781 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      841 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      901 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      961 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1021 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1081 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1141 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1201 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1261 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1321 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1381 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1441 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1501 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1561 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1621 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1681 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1741 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1801 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1861 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1921 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
     1981 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     2041 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     2101 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
     2161 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
     2221 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     2281 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     2341 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     2401 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     2461 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     2521 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     2581 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     2641 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     2701 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt
     2761 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     2821 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     2881 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     2941 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
     3001 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
     3061 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
     3121 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
     3181 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
     3241 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
     3301 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
     3361 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
     3421 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc
     3481 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     3541 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     3601 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     3661 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     3721 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
     3781 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
     3841 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
     3901 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     3961 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     4021 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     4081 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     4141 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     4201 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc
     4261 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
     4321 cggatcggca ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta
     4381 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     4441 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     4501 attacaggga cagcagagat ccagtttggt taattaaggt accgagggcc tatttcccat
     4561 gattccttca tatttgcata tacgatacaa ggctgttaga gagataatta gaattaattt
     4621 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
     4681 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
     4741 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgtggtgaa
     4801 aggcactata acagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
     4861 aacttgaaaa agtggcaccg agtcggtgct tttttgaatt cgctagctag gtcttgaaag
     4921 gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
     4981 agaagttggg gggaggggtc ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa
     5041 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
     5101 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
     5161 aggaccggtt ctagagcgct gccaccatgg acaagaagta cagcatcggc ctggacatcg
     5221 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat
     5281 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc
     5341 tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat
     5401 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca
     5461 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga
     5521 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt
     5581 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc
     5641 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg
     5701 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct
     5761 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc
     5821 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg
     5881 agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact
     5941 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg
     6001 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg
     6061 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga
     6121 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc
     6181 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct
     6241 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt
     6301 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga
     6361 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc
     6421 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat
     6481 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg
     6541 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa
     6601 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca
     6661 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca
     6721 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg
     6781 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc
     6841 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa
     6901 tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg
     6961 gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa
     7021 acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga
     7081 tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga
     7141 agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg
     7201 acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa
     7261 acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc
     7321 aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg
     7381 ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg
     7441 gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga
     7501 agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg
     7561 gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt
     7621 acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc
     7681 ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca
     7741 tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct
     7801 ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga
     7861 ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg
     7921 ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg
     7981 cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg
     8041 aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt
     8101 tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg
     8161 tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg
     8221 actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg
     8281 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc
     8341 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga
     8401 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag
     8461 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc
     8521 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg
     8581 gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg
     8641 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa
     8701 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa
     8761 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga
     8821 gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat
     8881 atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata
     8941 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc
     9001 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt
     9061 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc
     9121 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg
     9181 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca
     9241 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac aagcgacctg
     9301 ccgccacaaa gaaggctgga caggctaaga agaagaaaga ttacaaagac gatgacgata
     9361 agggatccgg cgcaacaaac ttctctctgc tgaaacaagc cggagatgtc gaagagaatc
     9421 ctggaccgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     9481 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     9541 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     9601 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     9661 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     9721 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     9781 aggagcccgc gtggttcctg gccaccgtcg gagtctcgcc cgaccaccag ggcaagggtc
     9841 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     9901 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     9961 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
    10021 cctgaacgcg ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
    10081 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
    10141 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
    10201 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
    10261 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
    10321 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
    10381 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
    10441 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
    10501 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
    10561 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
    10621 cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
    10681 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
    10741 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
    10801 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
    10861 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
    10921 gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
    10981 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
    11041 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
    11101 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
    11161 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
    11221 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
    11281 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
    11341 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
    11401 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
    11461 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
    11521 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
    11581 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
    11641 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
    11701 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
    11761 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
    11821 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
    11881 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
    11941 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
    12001 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
    12061 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
    12121 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
    12181 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
    12241 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
    12301 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
    12361 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
    12421 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
    12481 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg
    12541 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
    12601 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
    12661 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
    12721 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
    12781 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
    12841 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
    12901 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
    12961 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcg
//
