LOCUS       sequence-340304-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A knockout vector for the dog Egfr..
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: LentiCRISPRv2-puro-dEgfr
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    333..450
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        501..749
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     501..521
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     672..691
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        771..846
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        908..1119
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      1138..1147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1144..5247
                     /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             1144..5247
                     /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             5248..5295
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5296..5319
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5329..5385
                     /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             5386..5982
                     /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     5879..5899
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5998..6586
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6051..6071)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6469..6480)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6658..6891
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6917..6934)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6923..7147
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7193..7621
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7280..7299)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7490..7511
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7630..7650)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7635..7964
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7815..7950
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7877..7896
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8012..8059
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8078..8452
                     /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    8582..8703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8619..8638)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8673..8692
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8752..8768)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8752..8768)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8765..8787)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8776..8792
                     /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(8800..8806,8807..8824,8825..8830))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8845..8866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8983..9000)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9154..9742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9234..9253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9913..10704,10705..10773))
                     /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     10536..10555
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10774..10878)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10953..10972)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        11144..11523
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11525..11723
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11678..11698
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11741..11921
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11968..12093
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12586..12819
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             join(13004..13013,1..35)
                     /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"
ORIGIN
        1 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
       61 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
      121 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
      181 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
      241 gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa
      301 ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
      361 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
      421 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
      481 gtttggttaa ttaaggtacc gagggcctat ttcccatgat tccttcatat ttgcatatac
      541 gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag
      601 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
      661 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
      721 tatatatctt gtggaaagga cgaaacaccg cccaggaggc ggcacacgtg gttttagagc
      781 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
      841 cggtgctttt ttgaattcgc tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc
      901 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
      961 aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     1021 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     1081 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg accggttcta gagcgctgcc
     1141 accatggaca agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc
     1201 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac
     1261 cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc
     1321 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc
     1381 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
     1441 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc
     1501 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga
     1561 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc
     1621 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
     1681 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac
     1741 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc
     1801 agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga
     1861 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
     1921 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
     1981 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc
     2041 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc
     2101 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg
     2161 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac
     2221 gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc
     2281 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg
     2341 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg
     2401 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag
     2461 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac
     2521 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag
     2581 gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat
     2641 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
     2701 gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc
     2761 ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg
     2821 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa
     2881 atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa
     2941 attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc
     3001 gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat
     3061 gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg
     3121 ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc
     3181 ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac
     3241 gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc
     3301 ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag
     3361 acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc
     3421 gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag
     3481 agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac
     3541 cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg
     3601 cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac
     3661 catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga
     3721 agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg
     3781 aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat
     3841 ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga
     3901 cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg
     3961 aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag
     4021 tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac
     4081 aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa
     4141 aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg
     4201 aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac
     4261 agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag
     4321 cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat
     4381 tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag
     4441 gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg
     4501 atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg
     4561 gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt
     4621 gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc
     4681 gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct
     4741 aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa
     4801 ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc
     4861 agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg
     4921 gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga
     4981 gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat
     5041 aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga
     5101 gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc
     5161 aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg
     5221 atcgacctgt ctcagctggg aggcgacaag cgacctgccg ccacaaagaa ggctggacag
     5281 gctaagaaga agaaagatta caaagacgat gacgataagg gatccggcgc aacaaacttc
     5341 tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccgaccga gtacaagccc
     5401 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     5461 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     5521 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     5581 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     5641 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     5701 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     5761 accgtcggag tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     5821 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     5881 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     5941 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaacgcgtta agtcgacaat
     6001 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     6061 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     6121 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     6181 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     6241 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     6301 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     6361 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     6421 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     6481 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     6541 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
     6601 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     6661 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     6721 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6781 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6841 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt
     6901 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     6961 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     7021 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     7081 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     7141 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     7201 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     7261 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     7321 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     7381 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     7441 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     7501 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     7561 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     7621 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     7681 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     7741 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     7801 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     7861 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     7921 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     7981 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
     8041 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
     8101 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
     8161 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
     8221 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
     8281 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
     8341 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
     8401 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct
     8461 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     8521 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     8581 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     8641 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     8701 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     8761 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     8821 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     8881 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     8941 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     9001 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     9061 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     9121 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     9181 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     9241 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     9301 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     9361 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     9421 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     9481 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     9541 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     9601 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     9661 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     9721 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     9781 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     9841 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     9901 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     9961 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    10021 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
    10081 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
    10141 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    10201 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
    10261 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
    10321 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
    10381 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
    10441 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
    10501 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
    10561 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
    10621 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
    10681 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
    10741 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
    10801 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
    10861 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
    10921 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
    10981 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
    11041 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
    11101 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
    11161 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
    11221 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
    11281 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
    11341 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
    11401 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
    11461 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
    11521 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
    11581 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
    11641 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
    11701 cggtgggagg tctatataag cagcgcgttt tgcctgtact gggtctctct ggttagacca
    11761 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
    11821 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
    11881 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac
    11941 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg
    12001 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga
    12061 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg
    12121 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata
    12181 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca
    12241 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa
    12301 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata
    12361 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc
    12421 gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
    12481 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
    12541 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
    12601 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
    12661 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
    12721 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
    12781 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
    12841 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
    12901 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
    12961 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aac
//
