LOCUS       sequence-340299-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A knockout vector for the dog Hbegf..
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-dHbegf
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(67..86)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        258..637
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        639..837
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     792..812
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             855..1035
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1082..1207
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1700..1933
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2118..2162
                     /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    2460..2577
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2628..2876
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2628..2648
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2799..2818
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2898..2973
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3035..3246
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3265..3274
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3271..7374
                     /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             3271..7374
                     /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             7375..7422
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7423..7446
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7456..7512
                     /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             7513..8109
                     /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     8006..8026
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8125..8713
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8178..8198)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8596..8607)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8785..9018
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9044..9061)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9050..9274
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9320..9748
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9407..9426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9617..9638
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9757..9777)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9762..10091
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9942..10077
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10004..10023
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10139..10186
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10205..10579
                     /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    10709..10830
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10746..10765)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10800..10819
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10879..10895)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10879..10895)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10892..10914)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10903..10919
                     /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(10927..10933,10934..10951,10952..10957))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10972..10993
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11110..11127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11281..11869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11361..11380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12040..12831,12832..12900))
                     /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     12663..12682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12901..13005)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
       61 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
      121 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
      181 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
      241 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
      301 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      361 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      421 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      481 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      541 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      601 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      661 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      721 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      781 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagcgc
      841 gttttgcctg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
      901 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
      961 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     1021 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     1081 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     1141 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     1201 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     1261 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     1321 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     1381 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     1441 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     1501 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     1561 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     1621 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     1681 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     1741 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     1801 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     1861 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     1921 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     1981 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     2041 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     2101 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     2161 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     2221 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     2281 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca
     2341 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag
     2401 tgaacggatc ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt catccacaat
     2461 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     2521 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg
     2581 gtttattaca gggacagcag agatccagtt tggttaatta aggtaccgag ggcctatttc
     2641 ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata attagaatta
     2701 atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc
     2761 ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa
     2821 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgctc
     2881 ccaccgaatc cacggacgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
     2941 tatcaacttg aaaaagtggc accgagtcgg tgcttttttg aattcgctag ctaggtcttg
     3001 aaaggagtgg gaattggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     3061 cccgagaagt tggggggagg ggtcggcaat tgatccggtg cctagagaag gtggcgcggg
     3121 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     3181 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     3241 acacaggacc ggttctagag cgctgccacc atggacaaga agtacagcat cggcctggac
     3301 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
     3361 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
     3421 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
     3481 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
     3541 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
     3601 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
     3661 aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac
     3721 ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc
     3781 gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag
     3841 acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc
     3901 atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc
     3961 ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc
     4021 aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc
     4081 tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt
     4141 ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc
     4201 gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag
     4261 gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt
     4321 ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa
     4381 gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc
     4441 gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc
     4501 ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac
     4561 ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac
     4621 tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag
     4681 gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc
     4741 ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag
     4801 cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg
     4861 accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac
     4921 ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag
     4981 aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc
     5041 ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag
     5101 gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag
     5161 atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag
     5221 ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc
     5281 cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac
     5341 agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa
     5401 gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc
     5461 cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg
     5521 atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc
     5581 cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag
     5641 ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag
     5701 ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc
     5761 aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac
     5821 tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg
     5881 ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag
     5941 ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa
     6001 ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac
     6061 gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc
     6121 cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc
     6181 cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac
     6241 gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac
     6301 ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc
     6361 aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt
     6421 accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg
     6481 gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc
     6541 caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc
     6601 ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag
     6661 tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa
     6721 aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa
     6781 agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg
     6841 aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg
     6901 aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc
     6961 aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag
     7021 gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc
     7081 gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg
     7141 ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc
     7201 cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc
     7261 atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag
     7321 agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaagcga
     7381 cctgccgcca caaagaaggc tggacaggct aagaagaaga aagattacaa agacgatgac
     7441 gataagggat ccggcgcaac aaacttctct ctgctgaaac aagccggaga tgtcgaagag
     7501 aatcctggac cgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     7561 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     7621 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     7681 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     7741 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     7801 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     7861 cccaaggagc ccgcgtggtt cctggccacc gtcggagtct cgcccgacca ccagggcaag
     7921 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     7981 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     8041 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     8101 ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     8161 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     8221 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     8281 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     8341 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     8401 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     8461 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     8521 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     8581 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     8641 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     8701 gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     8761 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     8821 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     8881 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     8941 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     9001 tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     9061 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     9121 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     9181 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     9241 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     9301 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     9361 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     9421 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     9481 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     9541 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     9601 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     9661 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     9721 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     9781 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     9841 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     9901 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     9961 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
    10021 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
    10081 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
    10141 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
    10201 aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc
    10261 ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
    10321 cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc
    10381 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
    10441 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
    10501 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
    10561 ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga
    10621 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
    10681 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
    10741 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
    10801 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
    10861 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
    10921 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
    10981 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
    11041 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
    11101 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
    11161 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
    11221 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
    11281 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
    11341 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
    11401 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
    11461 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
    11521 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
    11581 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
    11641 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
    11701 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
    11761 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
    11821 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
    11881 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
    11941 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
    12001 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
    12061 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
    12121 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
    12181 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
    12241 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
    12301 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
    12361 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
    12421 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
    12481 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
    12541 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
    12601 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
    12661 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
    12721 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
    12781 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
    12841 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
    12901 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
    12961 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttc
//
