LOCUS       sequence-419655-        9756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockout of DNAJC5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9756)
  AUTHORS   Wu S, Hernandez Villegas NC, Sirkis DW, Thomas-Wright I, 
            Wade-Martins R, Schekman R
  TITLE     Unconventional secretion of alpha-synuclein mediated by 
            palmitoylated DNAJC5 oligomers.
  JOURNAL   Elife. 2023 Jan 10;12:e85837. doi: 10.7554/eLife.85837.
  PUBMED    36626307
REFERENCE   2  (bases 1 to 9756)
  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: "2023-01-10"; volume: "12:e85837. doi"; pages: " 
            10.7554/eLife.85837"
COMMENT     Sequence Label: pX330-Venus-DNAJC5-Exon 4-gRNA
FEATURES             Location/Qualifiers
     source          1..9756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          205..432
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      444..453
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             453..518
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             525..545
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             570..4670
                     /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             570..4670
                     /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             4671..4718
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4746..4763)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4752..4959
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4968..5108
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4968..5097
                     /label=AAV2 ITR
     rep_origin      5183..5541
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5270..5289)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5480..5501
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5551..6050
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5586..5609)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5983..6002
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             join(6073..6075,6076..6078,6079..6792)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNAIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     ELVTAAGITLGMDELYK"
     primer_bind     complement(6379..6398)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(6906..6925)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7025..7047
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7085..7103)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7171..7275
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7276..7344,7345..8136)
                     /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(7494..7513)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8307..8895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8796..8815
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8957..9205
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8957..8977
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9128..9147
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9227..9302
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        9399..9684
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(9686..9756,1..204)
                     /label=chicken beta-actin promoter
ORIGIN
        1 tttatttttt aattattttg tgcagcgatg ggggcggggg ggggggggcg cgtgccaggc
       61 ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat
      121 cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat
      181 aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc
      241 tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
      301 agcgggcggg acggcccttc tcctccgggc tgtaattagc tgagcaagag gtaagggttt
      361 aagggatggt tggttggtgg ggtattaatg tttaattacc tggagcacct gcctgaaatc
      421 actttttttc aggttggacc ggtgccacca tggactataa ggaccacgac ggagactaca
      481 aggatcatga tattgattac aaagacgatg acgataagat ggccccaaag aagaagcgga
      541 aggtcggtat ccacggagtc ccagcagccg acaagaagta cagcatcggc ctggacatcg
      601 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat
      661 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc
      721 tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat
      781 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca
      841 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga
      901 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt
      961 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc
     1021 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg
     1081 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct
     1141 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc
     1201 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg
     1261 agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact
     1321 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg
     1381 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg
     1441 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga
     1501 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc
     1561 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct
     1621 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt
     1681 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga
     1741 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc
     1801 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat
     1861 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg
     1921 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa
     1981 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca
     2041 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca
     2101 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg
     2161 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc
     2221 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa
     2281 tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg
     2341 gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa
     2401 acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga
     2461 tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga
     2521 agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg
     2581 acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa
     2641 acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc
     2701 aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg
     2761 ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg
     2821 gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga
     2881 agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg
     2941 gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt
     3001 acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc
     3061 ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca
     3121 tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct
     3181 ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga
     3241 ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg
     3301 ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg
     3361 cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg
     3421 aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt
     3481 tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg
     3541 tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg
     3601 actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg
     3661 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc
     3721 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga
     3781 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag
     3841 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc
     3901 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg
     3961 gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg
     4021 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa
     4081 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa
     4141 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga
     4201 gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat
     4261 atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata
     4321 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc
     4381 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt
     4441 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc
     4501 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg
     4561 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca
     4621 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac aaaaggccgg
     4681 cggccacgaa aaaggccggc caggcaaaaa agaaaaagta agaattccta gagctcgctg
     4741 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     4801 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     4861 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     4921 gggggaggat tgggaagaga atagcaggca tgctggggag cggccgcagg aacccctagt
     4981 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     5041 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     5101 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     5161 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5221 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     5281 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     5341 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     5401 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5461 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     5521 tcgggctatt cttttgattt aaattctacc gggtagggga ggcgcttttc ccaaggcagt
     5581 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc
     5641 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
     5701 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
     5761 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
     5821 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
     5881 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc
     5941 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa
     6001 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc
     6061 caagcttcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     6121 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     6181 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
     6241 ccctggccca ccctcgtgac caccctgggc tacggcctgc agtgcttcgc ccgctacccc
     6301 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     6361 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     6421 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     6481 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     6541 aagcagaaga acgccatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     6601 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     6661 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     6721 gatcacatgg tcctgctgga gttagtgacc gccgccggga tcactctcgg catggacgag
     6781 ctgtacaagt aataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     6841 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     6901 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     6961 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     7021 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     7081 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     7141 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     7201 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     7261 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     7321 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     7381 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     7441 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     7501 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     7561 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     7621 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     7681 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     7741 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     7801 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     7861 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     7921 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     7981 ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     8041 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     8101 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     8161 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     8221 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     8281 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     8341 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     8401 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     8461 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     8521 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     8581 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     8641 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     8701 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     8761 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     8821 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     8881 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     8941 ccttttgctc acatgtgagg gcctatttcc catgattcct tcatatttgc atatacgata
     9001 caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca
     9061 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
     9121 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
     9181 tatcttgtgg aaaggacgaa acaccggagg ccgcagaaga caaacagttt tagagctaga
     9241 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
     9301 gcttttttgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg tttttagcgc
     9361 gtgcgccaat tctgcagaca aatggctcta gaggtacccg ttacataact tacggtaaat
     9421 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaatagt aacgccaata
     9481 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     9541 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     9601 gcctggcatt gtgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     9661 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
     9721 atctcccccc cctccccacc cccaattttg tattta
//
