LOCUS       sequence-362121-        9300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cPLA2-KO in HeLa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9300)
  AUTHORS   Watabe T, Yamahira S, Takakura K, Thumkeo D, Narumiya S, Matsuda M, 
            Terai K
  TITLE     Calcium transients trigger switch-like discharge of prostaglandin 
            E(2) in an extracellular signal-regulated kinase-dependent manner.
  JOURNAL   eLife. 2024 Jan 26;12:RP86727. doi: 10.7554/eLife.86727.
  PUBMED    38276879
REFERENCE   2  (bases 1 to 9300)
  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: "eLife."; 
            date: "2024-01-26"; volume: "12:RP86727. doi"; pages: " 
            10.7554/eLife.86727"
COMMENT     Sequence Label: pX459-homo_PLA2G4A-2
FEATURES             Location/Qualifiers
     source          1..9300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..283
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     35..55
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     206..225
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        305..380
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        477..762
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          1165..1392
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1404..1413
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1413..1478
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1485..1505
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1530..5630
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1530..5630
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5631..5678
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5694..5747
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5748..6347
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5748..5767)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(6375..6392)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6381..6588
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6597..6737
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6597..6726
                     /label=AAV2 ITR
     rep_origin      6812..7267
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6899..6918)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7109..7130
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7284..7303)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7403..7425
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7463..7481)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7549..7653
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7654..7722,7723..8514)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7872..7891)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8685..9273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9174..9193
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttcctggcct tttgctggcc ttttgctcac atgtgagggc ctatttccca tgattccttc
       61 atatttgcat atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa
      121 cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc
      181 agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt
      241 cgatttcttg gctttatata tcttgtggaa aggacgaaac accgcaccac taccgtaaac
      301 ttgtgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
      361 aagtggcacc gagtcggtgc ttttttgttt tagagctaga aatagcaagt taaaataagg
      421 ctagtccgtt tttagcgcgt gcgccaattc tgcagacaaa tggctctaga ggtacccgtt
      481 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      541 tcaatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      601 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      661 aatgacggta aatggcccgc ctggcattgt gcccagtaca tgaccttatg ggactttcct
      721 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
      781 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
      841 ttttaattat tttgtgcagc gatgggggcg gggggggggg ggngggngcg nnccngggng
      901 gggngggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggbvd
      961 kgggggggbg vbbbgddggg gggngggggg gggggggggg gggggggngn gnsgssgggg
     1021 ggggsggggs ggggsgrggg gsggggsggg gcgaggcgga gaggtgcggc ggcagccaat
     1081 cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat
     1141 aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc
     1201 tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
     1261 agcgggcggg acggcccttc tcctccgggc tgtaattagc tgagcaagag gtaagggttt
     1321 aagggatggt tggttggtgg ggtattaatg tttaattacc tggagcacct gcctgaaatc
     1381 actttttttc aggttggacc ggtgccacca tggactataa ggaccacgac ggagactaca
     1441 aggatcatga tattgattac aaagacgatg acgataagat ggccccaaag aagaagcgga
     1501 aggtcggtat ccacggagtc ccagcagccg acaagaagta cagcatcggc ctggacatcg
     1561 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat
     1621 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc
     1681 tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat
     1741 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca
     1801 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga
     1861 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt
     1921 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc
     1981 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg
     2041 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct
     2101 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc
     2161 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg
     2221 agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact
     2281 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg
     2341 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg
     2401 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga
     2461 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc
     2521 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct
     2581 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt
     2641 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga
     2701 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc
     2761 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat
     2821 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg
     2881 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa
     2941 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca
     3001 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca
     3061 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg
     3121 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc
     3181 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa
     3241 tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg
     3301 gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa
     3361 acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga
     3421 tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga
     3481 agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg
     3541 acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa
     3601 acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc
     3661 aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg
     3721 ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg
     3781 gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga
     3841 agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg
     3901 gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt
     3961 acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc
     4021 ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca
     4081 tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct
     4141 ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga
     4201 ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg
     4261 ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg
     4321 cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg
     4381 aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt
     4441 tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg
     4501 tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg
     4561 actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg
     4621 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc
     4681 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga
     4741 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag
     4801 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc
     4861 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg
     4921 gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg
     4981 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa
     5041 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa
     5101 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga
     5161 gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat
     5221 atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata
     5281 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc
     5341 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt
     5401 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc
     5461 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg
     5521 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca
     5581 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac aaaaggccgg
     5641 cggccacgaa aaaggccggc caggcaaaaa agaaaaagga attcggcagt ggagagggca
     5701 gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc tggcccaatg accgagtaca
     5761 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     5821 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     5881 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     5941 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     6001 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     6061 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     6121 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     6181 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     6241 cccacaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     6301 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagaa ttctaactag
     6361 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     6421 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     6481 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     6541 ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagc ggccgcagga
     6601 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     6661 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     6721 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     6781 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     6841 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     6901 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6961 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     7021 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     7081 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     7141 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     7201 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     7261 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     7321 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     7381 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     7441 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     7501 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     7561 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     7621 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     7681 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     7741 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     7801 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     7861 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     7921 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     7981 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     8041 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     8101 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     8161 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     8221 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     8281 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     8341 gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca
     8401 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     8461 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     8521 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     8581 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     8641 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     8701 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     8761 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     8821 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     8881 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     8941 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     9001 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     9061 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     9121 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     9181 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     9241 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
//
