LOCUS       sequence-340303-       13498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A knockout vector for the dog Ereg..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13498)
  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 13498)
  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-hyg2-dEreg
FEATURES             Location/Qualifiers
     source          1..13498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    283..400
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        451..699
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     451..471
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     622..641
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        721..796
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        858..1069
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      1088..1097
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1094..5197
                     /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             1094..5197
                     /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             5198..5245
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5246..5269
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    5309..5876
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5470..5487)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5697..5716
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5915..6934
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     LTR             7093..7326
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7352..7369)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7358..7582
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7628..8056
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7715..7734)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7925..7946
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8065..8085)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8070..8399
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8250..8385
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8312..8331
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8447..8494
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8513..8887
                     /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    9017..9138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9054..9073)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9108..9127
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9187..9203)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9187..9203)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9200..9222)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9211..9227
                     /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(9235..9241,9242..9259,9260..9265))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9280..9301
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9418..9435)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9589..10177)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9669..9688)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10348..11139,11140..11208))
                     /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     10971..10990
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11209..11313)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(11388..11407)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        11579..11958
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11960..12158
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     12113..12133
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             12176..12356
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    12403..12528
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    13021..13254
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             13439..13483
                     /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 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
       61 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
      121 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
      181 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
      241 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
      301 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
      361 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
      421 cagagatcca gtttggttaa ttaaggtacc gagggcctat ttcccatgat tccttcatat
      481 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
      541 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
      601 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
      661 ttcttggctt tatatatctt gtggaaagga cgaaacaccg gatagaagac aacccacgtg
      721 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
      781 ggcaccgagt cggtgctttt ttgaattcgc tagctaggtc ttgaaaggag tgggaattgg
      841 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
      901 aggggtcggc aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
      961 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     1021 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg accggttcta
     1081 gagcgctgcc accatggaca agaagtacag catcggcctg gacatcggca ccaactctgt
     1141 gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg
     1201 caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg
     1261 cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa
     1321 gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag
     1381 cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca
     1441 ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta
     1501 ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct
     1561 ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc
     1621 cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt
     1681 cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact
     1741 gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg
     1801 cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt
     1861 cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga
     1921 caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct
     1981 gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc
     2041 cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa
     2101 agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa
     2161 gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat
     2221 caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga
     2281 ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct
     2341 gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa
     2401 ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc
     2461 caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg
     2521 gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac
     2581 caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga
     2641 gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa
     2701 gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa
     2761 ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga
     2821 ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga
     2881 tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct
     2941 ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct
     3001 gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata
     3061 caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg
     3121 caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct
     3181 gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca
     3241 gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg
     3301 catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc
     3361 cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa
     3421 cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct
     3481 gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct
     3541 gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta
     3601 cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt
     3661 gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt
     3721 gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa
     3781 gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt
     3841 catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga
     3901 ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat
     3961 caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg
     4021 cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc
     4081 cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta
     4141 cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta
     4201 cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga
     4261 gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa
     4321 gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa
     4381 aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag
     4441 cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag
     4501 ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa
     4561 actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa
     4621 gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat
     4681 caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc
     4741 tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct
     4801 gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca
     4861 gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt
     4921 ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa
     4981 gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac
     5041 caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta
     5101 caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta
     5161 cgagacacgg atcgacctgt ctcagctggg aggcgacaag cgacctgccg ccacaaagaa
     5221 ggctggacag gctaagaaga agaaagatta caaagacgat gacgataagg gctctagagg
     5281 atcccggttt ccctctagcg ggatcaattc cgcccccccc ccctaacgtt actggccgaa
     5341 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     5401 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     5461 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     5521 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     5581 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     5641 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     5701 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     5761 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     5821 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt
     5881 gccacaaccc gtaccaaaga tggatagatc cggaaagcct gaactcaccg cgacgtctgt
     5941 cgagaagttt ctgatcgaaa agttcgacag cgtgtccgac ctgatgcagc tctcggaggg
     6001 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     6061 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc
     6121 gctcccgatt ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat
     6181 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     6241 tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag
     6301 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     6361 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     6421 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     6481 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     6541 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     6601 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     6661 gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct
     6721 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     6781 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     6841 cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     6901 acgccccagc actcgtccga gggcaaagga atagagggcc tgctgccggc tctgcggcct
     6961 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     7021 cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     7081 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt
     7141 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     7201 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     7261 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     7321 ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     7381 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     7441 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     7501 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     7561 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     7621 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     7681 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     7741 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     7801 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     7861 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     7921 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     7981 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     8041 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     8101 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     8161 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     8221 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     8281 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     8341 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     8401 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc
     8461 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa
     8521 gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg
     8581 gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg
     8641 ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct
     8701 ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc
     8761 cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg
     8821 gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca
     8881 ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     8941 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     9001 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     9061 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     9121 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     9181 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     9241 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     9301 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     9361 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     9421 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     9481 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     9541 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     9601 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     9661 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     9721 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     9781 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     9841 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     9901 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     9961 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
    10021 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
    10081 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
    10141 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
    10201 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
    10261 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
    10321 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
    10381 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
    10441 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
    10501 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
    10561 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
    10621 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
    10681 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
    10741 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
    10801 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
    10861 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
    10921 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
    10981 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
    11041 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
    11101 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
    11161 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
    11221 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
    11281 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
    11341 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
    11401 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
    11461 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
    11521 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
    11581 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
    11641 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
    11701 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
    11761 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
    11821 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
    11881 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
    11941 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
    12001 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
    12061 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
    12121 ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc
    12181 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
    12241 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
    12301 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
    12361 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
    12421 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
    12481 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
    12541 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
    12601 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
    12661 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
    12721 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
    12781 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
    12841 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
    12901 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
    12961 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
    13021 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
    13081 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
    13141 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
    13201 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
    13261 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
    13321 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
    13381 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
    13441 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaac
//
