LOCUS       sequence-233965-       14578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Valcyrie construct for CRISPR-mediated lox excision in Tribolium 
            castaneum (high).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14578)
  AUTHORS   Gilles AF, Schinko JB, Schacht MI, Enjolras C, Averof M
  TITLE     Clonal analysis by tunable CRISPR-mediated excision.
  JOURNAL   Development. 2019 Jan 4;146(1). pii: dev.170969. doi: 
            10.1242/dev.170969.
  PUBMED    30552128
REFERENCE   2  (bases 1 to 14578)
  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: 
            "Development."; date: "2019-01-4"; volume: "146(1). pii: dev.170969.
            doi"; pages: " 10.1242/dev.170969"
COMMENT     Sequence Label: pBac(3xP3-gTc'v; Valcyrie.LR)
FEATURES             Location/Qualifiers
     source          1..14578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        661..736
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             1651..1671
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      1849..1858
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1855..5958
                     /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             1855..5958
                     /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             5965..5985
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_RNA        6801..6876
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     polyA_signal    6911..7032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6922..6941)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6976..6995
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   9828..9890
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(10594..10611)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10594..10610)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10819..10838)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10938..10960
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(10998..11016)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        11084..11188
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11189..11257,11258..12049)
                     /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(11407..11426)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12220..12808
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12709..12728
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12962..12979
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13096..13117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13132..13137,13138..13155,13156..13162)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13170..13186
                     /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)."
     primer_bind     13175..13197
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13194..13210
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13194..13210
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   13391..13425
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 gccaagtaag tatggaataa aaatcctcat gatgtgtgac agtggtacga agtatatgat
       61 aaatggaatg ccttatttgg gaagaggaac acagaccaac ggagtaccac tcggtgaata
      121 ctacgtgaag gagttatcaa agcctgtgca cggtagttgt cgtaatatta cgtgtgacaa
      181 ttggttcacc tcaatccctt tggcaaaaaa cttactacaa gaaccgtata agttcgagat
      241 cggccggccc ttgcaaaaac aattcgttta cgtccgttaa cacgttctcg taatccaaaa
      301 ataaataatt tatggtcttt atcaaattaa tcgtgtagag aactgtatca tcaacaatta
      361 ataatttgta catgattatt gattatggat agtctagact aaacctttta cgaaatcaat
      421 gactaactgt tttatttatt tatttatgtg ttaatattaa ttgagataat tttaagttta
      481 atgctaataa tgtaacaact tcttattaaa tagtaaacac aaacaaattg ataaaaacca
      541 gtcatattct aaaattaaaa aagttattgt ggaggttaca tctcactaac gctggtcgcc
      601 cgtcgctacg ttcatataca gattgttgag aagggcaagt gtacgtaata acttcgtata
      661 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
      721 ggcaccgagt cggtgctttt ttggccggcc gaattcgaat ggccatggga cgtcgacctg
      781 aggtaattat aacccgggcc ctatatatgg atcccaacct tcaataaaaa attgttcttt
      841 atttattatt aatcactatt atttaggcat cttctgcaaa agtaattgat tataaacgat
      901 aaaagtaaaa aaaaggatta atctaaaaca ttttacttaa agacctatta ataagtttaa
      961 aaagaaaaaa aataggaaaa ttgttttatt aaaattgcta tgacaatcca actttggcat
     1021 atgtatatga aatcaaaaca attcactact cattcataat attgccgaac tttaccaaat
     1081 taaaaataaa catgaaatag tcaacaactc attcgttatg taacgttttt tttttaaatg
     1141 cattaatata taatgattgt ttttattttg gaagttgaaa taaattaggg cacaaccagt
     1201 ttccactatt ctcctttatc tttacggaaa tagcatgaat aataccgaaa aggccgtttt
     1261 aaaaataaaa attcgagaaa attcaaacgc aaatcaatgg aaacttcgag atcattcggt
     1321 gaagttcgag acgaattccc cgtttcatat ataagcgcgg tctcgcggcg cgttgtcagt
     1381 gaatttgaat caagcgaagc gaaagcaaca aagctaagca agcgactcaa agcaaatttt
     1441 caaagcgact ttaacaaagt tgagtgaatt atttactagt gaattcaaag tcgtcgataa
     1501 ataagtttcc catttttatt ccccagccag gaagttagtt tcaatagttt tgtaatttca
     1561 acgaaactca tttgatttcg tactaatttt ccacatctct attttctgcc cgcagaataa
     1621 tccaaaatgc agatcgggga tcccacccca cccaagaaga agcgcaaggt ggaggacgat
     1681 cccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     1741 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     1801 tcccaacagt tgcggtcgac tctagaggat ccccgggatc caccggtcgc caccatggat
     1861 aagaaatact caataggctt agatatcggc acaaatagcg tcggatgggc ggtgatcact
     1921 gatgaatata aggttccgtc taaaaagttc aaggttctgg gaaatacaga ccgccacagt
     1981 atcaaaaaaa atcttatagg ggctctttta tttgacagtg gagagacagc ggaagcgact
     2041 cgtctcaaac ggacagctcg tagaaggtat acacgtcgga agaatcgtat ttgttatcta
     2101 caggagattt tttcaaatga gatggcgaaa gtagatgata gtttctttca tcgacttgaa
     2161 gagtcttttt tggtggaaga agacaagaag catgaacgtc atcctatttt tggaaatata
     2221 gtagatgaag ttgcttatca tgagaaatat ccaactatct atcatctgcg aaaaaaattg
     2281 gtagattcta ctgataaagc ggatttgcgc ttaatctatt tggccttagc gcatatgatt
     2341 aagtttcgtg gtcatttttt gattgaggga gatttaaatc ctgataatag tgatgtggac
     2401 aaactattta tccagttggt acaaacctac aatcaattat ttgaagaaaa ccctattaac
     2461 gcaagtggag tagatgctaa agcgattctt tctgcacgat tgagtaaatc aagacgatta
     2521 gaaaatctca ttgctcagct ccccggtgag aagaaaaatg gcttatttgg gaatctcatt
     2581 gctttgtcat tgggtttgac ccctaatttt aaatcaaatt ttgatttggc agaagatgct
     2641 aaattacagc tttcaaaaga tacttacgat gatgatttag ataatttatt ggcgcaaatt
     2701 ggagatcaat atgctgattt gtttttggca gctaagaatt tatcagatgc tattttactt
     2761 tcagatatcc taagagtaaa tactgaaata actaaggctc ccctatcagc ttcaatgatt
     2821 aaacgctacg atgaacatca tcaagacttg actcttttaa aagctttagt tcgacaacaa
     2881 cttccagaaa agtataaaga aatctttttt gatcaatcaa aaaacggata tgcaggttat
     2941 attgatgggg gagctagcca agaagaattt tataaattta tcaaaccaat tttagaaaaa
     3001 atggatggta ctgaggaatt attggtgaaa ctaaatcgtg aagatttgct gcgcaagcaa
     3061 cggacctttg acaacggctc tattccccat caaattcact tgggtgagct gcatgctatt
     3121 ttgagaagac aagaagactt ttatccattt ttaaaagaca atcgtgagaa gattgaaaaa
     3181 atcttgactt ttcgaattcc ttattatgtt ggtccattgg cgcgtggcaa tagtcgtttt
     3241 gcatggatga ctcggaagtc tgaagaaaca attaccccat ggaattttga agaagttgtc
     3301 gataaaggtg cttcagctca atcatttatt gaacgcatga caaactttga taaaaatctt
     3361 ccaaatgaaa aagtactacc aaaacatagt ttgctttatg agtattttac ggtttataac
     3421 gaattgacaa aggtcaaata tgttactgaa ggaatgcgaa aaccagcatt tctttcaggt
     3481 gaacagaaga aagccattgt tgatttactc ttcaaaacaa atcgaaaagt aaccgttaag
     3541 caattaaaag aagattattt caaaaaaata gaatgttttg atagtgttga aatttcagga
     3601 gttgaagata gatttaatgc ttcattaggt acctaccatg atttgctaaa aattattaaa
     3661 gataaagatt ttttggataa tgaagaaaat gaagatatct tagaggatat tgttttaaca
     3721 ttgaccttat ttgaagatag ggagatgatt gaggaaagac ttaaaacata tgctcacctc
     3781 tttgatgata aggtgatgaa acagcttaaa cgtcgccgtt atactggttg gggacgtttg
     3841 tctcgaaaat tgattaatgg tattagggat aagcaatctg gcaaaacaat attagatttt
     3901 ttgaaatcag atggttttgc caatcgcaat tttatgcagc tgatccatga tgatagtttg
     3961 acatttaaag aagacattca aaaagcacaa gtgtctggac aaggcgatag tttacatgaa
     4021 catattgcaa atttagctgg tagccctgct attaaaaaag gtattttaca gactgtaaaa
     4081 gttgttgatg aattggtcaa agtaatgggg cggcataagc cagaaaatat cgttattgaa
     4141 atggcacgtg aaaatcagac aactcaaaag ggccagaaaa attcgcgaga gcgtatgaaa
     4201 cgaatcgaag aaggtatcaa agaattagga agtcagattc ttaaagagca tcctgttgaa
     4261 aatactcaat tgcaaaatga aaagctctat ctctattatc tccaaaatgg aagagacatg
     4321 tatgtggacc aagaattaga tattaatcgt ttaagtgatt atgatgtcga tcacattgtt
     4381 ccacaaagtt tccttaaaga cgattcaata gacaataagg tcttaacgcg ttctgataaa
     4441 aatcgtggta aatcggataa cgttccaagt gaagaagtag tcaaaaagat gaaaaactat
     4501 tggagacaac ttctaaacgc caagttaatc actcaacgta agtttgataa tttaacgaaa
     4561 gctgaacgtg gaggtttgag tgaacttgat aaagctggtt ttatcaarcg ccaattggtt
     4621 gaaactcgcc aaatcactaa gcatgtggca caaattttgg atagtcgcat gaatactaaa
     4681 tacgatgaaa atgataaact tattcgagag gttaaagtga ttaccttaaa atctaaatta
     4741 gtttctgact tccgaaaaga tttccaattc tataaagtac gtgagattaa caattaccat
     4801 catgcccatg atgcgtatct aaatgccgtc gttggaactg ctttgattaa gaaatatcca
     4861 aaacttgaat cggagtttgt ctatggtgat tataaagttt atgatgttcg taaaatgatt
     4921 gctaagtctg agcaagaaat aggcaaagca accgcaaaat atttctttta ctctaatatc
     4981 atgaacttct tcaaaacaga aattacactt gcaaatggag agattcgcaa acgccctcta
     5041 atcgaaacta atggggaaac tggagaaatt gtctgggata aagggcgaga ttttgccaca
     5101 gtgcgcaaag tattgtccat gccccaagtc aatattgtca agaaaacaga agtacagaca
     5161 ggcggattct ccaaggagtc aattttacca aaaagaaatt cggacaagct tattgctcgt
     5221 aaaaaagact gggatccaaa aaaatatggt ggttttgata gtccaacggt agcttattca
     5281 gtcctagtgg ttgctaaggt ggaaaaaggg aaatcgaaga agttaaaatc cgttaaagag
     5341 ttactaggga tcacaattat ggaaagaagt tcctttgaaa aaaatccgat tgacttttta
     5401 gaagctaaag gatataagga agttaaaaaa gacttaatca ttaaactacc taaatatagt
     5461 ctttttgagt tagaaaacgg tcgtaaacgg atgctggcta gtgccggaga attacaaaaa
     5521 ggaaatgagc tggctctgcc aagcaaatat gtgaattttt tatatttagc tagtcattat
     5581 gaaaagttga agggtagtcc agaagataac gaacaaaaac aattgtttgt ggagcagcat
     5641 aagcattatt tagatgagat tattgagcaa atcagtgaat tttctaagcg tgttatttta
     5701 gcagatgcca atttagataa agttcttagt gcatataaca aacatagaga caaaccaata
     5761 cgtgaacaag cagaaaatat tattcattta tttacgttga cgaatcttgg agctcccgct
     5821 gcttttaaat attttgatac aacaattgat cgtaaacgat atacgtctac aaaagaagtt
     5881 ttagatgcca ctcttatcca tcaatccatc actggtcttt atgaaacacg cattgatttg
     5941 agtcagctag gaggtgacgg ttctcccaag aagaagagga aagtctcgag cggtggagct
     6001 gcaggatgag cggccgcctg cagctggcgc catcgatacg cgtacgtcgc gaccgcggac
     6061 atgtacagag ctcgagatga tcggtttgtg ttcctactta gttaattgtg atagtttatt
     6121 tttgtaaata atagtacgta aagttacttt agttataata aagtagatag tgactgcatt
     6181 aagtttaaag actgattttt gtagattatt taaaaatttt taataaatta tttaatttaa
     6241 agataaaaag ttgtttcatt ttaaaataat aaaaaactta ccgcattagt cgtggccttg
     6301 gcagtattaa gcgcaaacgg gctctccttc gtatgcccat caggaatata gaaaaaaaca
     6361 actggctgct cactagtggc cacgtgggcc gtgcacctta agcttggcgc gccgggtgtt
     6421 attattatta cttgtgacct ggtgataagt gcttcaagta ctagagattg agtcactgtt
     6481 caaggattat ttaaaaacac aacacgcaga tttatcagtt accttatgaa caaatgaata
     6541 tctatttttg tttcaaaact gttttcatta cgacaaatta atatttcttc aactaaataa
     6601 taaaaatttt aaataagcgc cattacatac aaatatacga atcattacga acgtttgtgt
     6661 tttgcgatcg attgtaaccg accaatcaaa ataaagcggg aaatttcaaa tctcttatat
     6721 ctcactacta cgtatcggtt gccgctaact acatatacaa ggcaggtaga agcaccgttc
     6781 gatcgcaata acttcgtata gttttagagc tagaaatagc aagttaaaat aaggctagtc
     6841 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttggcgcgcc gtacgcgtat
     6901 cgataagctt taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     6961 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     7021 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     7081 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcta
     7141 gagtcgcggc cgccttaacg aaataattaa ttgtttccat tcgacacgtt attggtgcta
     7201 ttcgccgacc cccaattgca caaatgactt ctcatagacg tagacaaggg ggggatgtag
     7261 gaccgtggta tcaaaaacgt cgataaattg aaaaggtcca cgaaaacttt ataccgatcg
     7321 ctgaaaaaca aaatcaaaca ttgcaaaaac cgtgttgaaa tcaatttcac cttaacgttg
     7381 accttaaata ctgatacccc gaagagcccc ctgttcctaa ctgcgaagaa ccaatcattc
     7441 tttgcaccat caaaacatga ttatctgaaa ttataaaatt taaacaaaaa caattgacgt
     7501 acggatttac taacaccgcc atttcgtaat cagcgagtct atgtccatca aaagtgttag
     7561 aatttggtgg ggctgactga acctgggttc gtccctgtaa aaggtgatca tgatagcccc
     7621 ttgcagggct ttgtgtgaga agcgcctctc gccgcgcgac attagagcct catggacctc
     7681 aactttaaaa atcgactcgt aaacctcgcg tctcttctcc aaatcgttca atttgtaacg
     7741 ctttaaagtc tccgcctcct ctttctccgc cagttctttt tgctcggtta aaagcttctc
     7801 gaccgctttt tggtacttcc cccagaaatt aaacccctcg agctccagcc ccggcgttcg
     7861 ctccagccac cgctggacca aatcggtcaa agacggctct ttctccgact tggcgatttg
     7921 ctcgagcgct ttttcgtcgt ttccgaacac tttcgagtaa ttctggttgt atttgacgcg
     7981 gttttcctgg cgcaccccga gcttattttc caggagcctg aactgcaggc tctggaaccc
     8041 ggaagcgggc cgcagataac accggaagtc catgaaatcg agcggtgtca tcgtttccag
     8101 aatcattact tggtccacca aaacctacaa ttgcgacagt tttatctcgt gaccgcaatt
     8161 tgttgcgaaa ttgtgaacct tgaggatcaa aacaacacga ttgaggcgtt tgaggatttc
     8221 caaagtttgc gactcttcta aaacgtcgct gaaaatgttg cggatcgagt ccagctcgta
     8281 gattatttgt ttgaaccata actcatacgc tggaattact tgttatagtt aattacggtt
     8341 aagtgggttt tttaccttga tgggtcacaa taaataagtg ctcgtcgtgc acgggttggt
     8401 tattttgctc gctgagtagt ctttgggctt ctaaaatctt gtccaacatc aagtattcac
     8461 cgtaaagcat cccacattct tcgctcagtt ggtcgccttc ttgggcttca cttaatttac
     8521 gattaattgt aaatggaata attaaattaa atcatactta cgagggtctc agtgggcaac
     8581 tcattacgac cgcggtaccg tcgactctag cggtaccccg attgtttagc ttgttcagct
     8641 gcgcttgttt atttgcttag ctttcgctta gcgacgtgtt cactttgctt gtttgaattg
     8701 aattgtcgct ccgtagacga agcgcctcta tttatactcc ggcggtcgag ggttcgaaat
     8761 cgataagctt ggatcctaat tgaattagct ctaattgaat tagtctctaa ttgaattaga
     8821 tccccgggcg agctcgaatt aaccattgtg ggaaccgtgc gatcaaacaa acgcgagata
     8881 ccggaagtac tgaaaaacag tcgctccagg ccagtgggaa catcgatgtt ttgttttgac
     8941 ggacccctta ctctcgtctc atataaaccg aagccagcta agatggtata cttattatca
     9001 tcttgtgatg aggatgcttc tatcaacgaa agtaccggta aaccgcaaat ggttatgtat
     9061 tataatcaaa ctaaaggcgg agtggacacg ctagaccaaa tgtgttctgt gatgacctgc
     9121 agtaggaaga cgaataggtg gcctatggca ttattgtacg gaatgataaa cattgcctgc
     9181 ataaattctt ttattatata cagccataat gtcagtagca agggagaaaa ggttcaaagt
     9241 cgcaaaaaat ttatgagaaa cctttacatg agcctgacgt catcgtttat gcgtaagcgt
     9301 ttagaagctc ctactttgaa gagatatttg cgcgataata tctctaatat tttgccaaat
     9361 gaagtgcctg gtacatcaga tgacagtact gaagagccag taatgaaaaa acgtacttac
     9421 tgtacttact gcccctctaa aataaggcga aaggcaaatg catcgtgcaa aaaatgcaaa
     9481 aaagttattt gtcgagagca taatattgat atgtgccaaa gttgtttctg actgactaat
     9541 aagtataatt tgtttctatt atgtataagt taagctaatt acttatttta taatacaaca
     9601 tgactgtttt taaagtacaa aataagttta tttttgtaaa agagagaatg tttaaaagtt
     9661 ttgttacttt atagaagaaa ttttgagttt ttgttttttt ttaataaata aataaacata
     9721 aataaattgt ttgttgaatt tattattagt atgtaagtgt aaatataata aaacttaata
     9781 tctattcaaa ttaataaata aacctcgata tacagaccga taaaacacat gcgtcaattt
     9841 tacgcatgat tatctttaac gtacgtcaca atatgattat ctttctaggg ttaaataata
     9901 gtttctaatt tttttattat tcagcctgct gtcgtgaata ccgtatatct caacgctgtc
     9961 tgtgagattg tcgtattcta gcctttttag tttttcgctc atcgacttga tattgtccga
    10021 cacattttcg tcgatttgcg ttttgatcaa agacttgagc agagacacgt taatcaactg
    10081 ttcaaattga tccatattaa cgatatcaac ccgatgcgta tatggtgcgt aaaatatatt
    10141 ttttaaccct cttatacttt gcactctgcg ttaatacgcg ttcgtgtaca gacgtaatca
    10201 tgttttcttt tttggataaa actcctactg agtttgacct catattagac cctcacaagt
    10261 tgcaaaacgt ggcatttttt accaatgaag aatttaaagt tattttaaaa aatttcatca
    10321 cagatttaaa gaagaaccaa aaattaaatt atttcaacag tttaatcgac cagttaatca
    10381 acgtgtacac agacgcgtcg gcaaaaaaca cgcagcccga cgtgttggct aaaattatta
    10441 aatcaacttg tgttatagtc acggatttgc cgtccaacgt gttcctcaaa aagttgaaga
    10501 ccaacaagtt tacggacact attaattatt tgattttgcc ccacttcatt ttgtgggatc
    10561 acaattttgt tatattttta aacaaagctt ggcactggcc gtcgttttac aacgtcgtga
    10621 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
    10681 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
    10741 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
    10801 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
    10861 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
    10921 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
    10981 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
    11041 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
    11101 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
    11161 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
    11221 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
    11281 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
    11341 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
    11401 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
    11461 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
    11521 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
    11581 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
    11641 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
    11701 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
    11761 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
    11821 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
    11881 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
    11941 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
    12001 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
    12061 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
    12121 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
    12181 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
    12241 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
    12301 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
    12361 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
    12421 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
    12481 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
    12541 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
    12601 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
    12661 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
    12721 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
    12781 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
    12841 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
    12901 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
    12961 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
    13021 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
    13081 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
    13141 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
    13201 cagctatgac atgattacga attcgagctc ggtacccggg gatcctctag agtcgacgct
    13261 cgcgcgactt ggtttgccat tctttagcgc gcgtcgcgtc acacagcttg gccacaatgt
    13321 ggtttttgtc aaacgaagat tctatgacgt gtttaaagtt taggtcgagt aaagcgcaaa
    13381 tcttttttaa ccctagaaag atagtctgcg taaaattgac gcatgcattc ttgaaatatt
    13441 gctctctctt tctaaatagc gcgaatccgt cgctgtgcat ttaggacatc tcagtcgccg
    13501 cttggagctc ccgtgaggcg tgcttgtcaa tgcggtaagt gtcactgatt ttgaactata
    13561 acgaccgcgt gagtcaaaat gacgcatgat tatcttttac gtgactttta agatttaact
    13621 catacgataa ttatattgtt atttcatgtt ctacttacgt gataacttat tatatatata
    13681 ttttcttgtt atagatatcg tgactaatat ataataaaat gggtagttct ttagacgatg
    13741 agcatatcct ctctgctctt ctgcaaagcg atgacgagct tgttggtgag gattctgaca
    13801 gtgaaatatc agatcacgta agtgaagatg acgtccagag cgatacagaa gaagcgttta
    13861 tagatgaggt acatgaagtg cagccaacgt caagcggtag tgaaatatta gacgaacaaa
    13921 atgttattga acaaccaggt tcttcattgg cttctaacag aatcttgacc ttgccacaga
    13981 ggactattag aggtaagaat aaacattgtt ggtcaacttc aaagtccacg aggcgtagcc
    14041 gagtctctgc actgaacatt gtcagatctc aaagaggtcc gacgcgtatg tgccgcaata
    14101 tatatgaccc acttttatgc ttcaaactat tttttactga tgagataatt tcggaaattg
    14161 taaaatggac aaatgctgag atatcattga aacgtcggga atctatgaca ggtgctacat
    14221 ttcgtgacac gaatgaagat gaaatctatg ctttctttgg tattctggta atgacagcag
    14281 tgagaaaaga taaccacatg tccacagatg acctctttga tcgatctttg tcaatggtgt
    14341 acgtctctgt aatgagtcgt gatcgttttg attttttgat acgatgtctt agaatggatg
    14401 acaaaagtat acggcccaca cttcgagaaa acgatgtatt tactcctgtt agaaaaatat
    14461 gggatctctt tatccatcag tgcatacaaa attacactcc aggggctcat ttgaccatag
    14521 atgaacagtt acttggtttt agaggacggt gtccgtttag gatgtatatc ccaaacaa
//
