LOCUS       sequence-234562-       14077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Valcyrie construct for CRISPR-mediated lox excision in Tribolium 
            castaneum (low).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14077)
  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 14077)
  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.Uni)
FEATURES             Location/Qualifiers
     source          1..14077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     371..388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    505..526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(541..546,547..564,565..571)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    579..595
                     /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     584..606
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     603..619
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     603..619
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   800..834
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     CDS             3137..3157
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      3335..3344
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3341..7444
                     /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             3341..7444
                     /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             7451..7471
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_RNA        complement(7906..7981)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     polyA_signal    8397..8518
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8408..8427)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8462..8481
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   11314..11376
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(12080..12097)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(12080..12096)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12305..12324)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12424..12446
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(12484..12502)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        12570..12674
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12675..12743,12744..13535)
                     /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(12893..12912)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(13706..14077,1..217)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
       61 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
      241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
      301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
      361 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
      421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
      481 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      541 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      601 cacaggaaac agctatgaca tgattacgaa ttcgagctcg gtacccgggg atcctctaga
      661 gtcgacgctc gcgcgacttg gtttgccatt ctttagcgcg cgtcgcgtca cacagcttgg
      721 ccacaatgtg gtttttgtca aacgaagatt ctatgacgtg tttaaagttt aggtcgagta
      781 aagcgcaaat cttttttaac cctagaaaga tagtctgcgt aaaattgacg catgcattct
      841 tgaaatattg ctctctcttt ctaaatagcg cgaatccgtc gctgtgcatt taggacatct
      901 cagtcgccgc ttggagctcc cgtgaggcgt gcttgtcaat gcggtaagtg tcactgattt
      961 tgaactataa cgaccgcgtg agtcaaaatg acgcatgatt atcttttacg tgacttttaa
     1021 gatttaactc atacgataat tatattgtta tttcatgttc tacttacgtg ataacttatt
     1081 atatatatat tttcttgtta tagatatcgt gactaatata taataaaatg ggtagttctt
     1141 tagacgatga gcatatcctc tctgctcttc tgcaaagcga tgacgagctt gttggtgagg
     1201 attctgacag tgaaatatca gatcacgtaa gtgaagatga cgtccagagc gatacagaag
     1261 aagcgtttat agatgaggta catgaagtgc agccaacgtc aagcggtagt gaaatattag
     1321 acgaacaaaa tgttattgaa caaccaggtt cttcattggc ttctaacaga atcttgacct
     1381 tgccacagag gactattaga ggtaagaata aacattgttg gtcaacttca aagtccacga
     1441 ggcgtagccg agtctctgca ctgaacattg tcagatctca aagaggtccg acgcgtatgt
     1501 gccgcaatat atatgaccca cttttatgct tcaaactatt ttttactgat gagataattt
     1561 cggaaattgt aaaatggaca aatgctgaga tatcattgaa acgtcgggaa tctatgacag
     1621 gtgctacatt tcgtgacacg aatgaagatg aaatctatgc tttctttggt attctggtaa
     1681 tgacagcagt gagaaaagat aaccacatgt ccacagatga cctctttgat cgatctttgt
     1741 caatggtgta cgtctctgta atgagtcgtg atcgttttga ttttttgata cgatgtctta
     1801 gaatggatga caaaagtata cggcccacac ttcgagaaaa cgatgtattt actcctgtta
     1861 gaaaaatatg ggatctcttt atccatcagt gcatacaaaa ttacactcca ggggctcatt
     1921 tgaccataga tgaacagtta cttggtttta gaggacggtg tccgtttagg atgtatatcc
     1981 caaacaagcc aagtaagtat ggaataaaaa tcctcatgat gtgtgacagt ggtacgaagt
     2041 atatgataaa tggaatgcct tatttgggaa gaggaacaca gaccaacgga gtaccactcg
     2101 gtgaatacta cgtgaaggag ttatcaaagc ctgtgcacgg tagttgtcgt aatattacgt
     2161 gtgacaattg gttcacctca atccctttgg caaaaaactt actacaagaa ccgtataagt
     2221 tcgagatcgg ccggccgaat tcgaatggcc atgggacgtc gacctgaggt aattataacc
     2281 cgggccctat atatggatcc caaccttcaa taaaaaattg ttctttattt attattaatc
     2341 actattattt aggcatcttc tgcaaaagta attgattata aacgataaaa gtaaaaaaaa
     2401 ggattaatct aaaacatttt acttaaagac ctattaataa gtttaaaaag aaaaaaaata
     2461 ggaaaattgt tttattaaaa ttgctatgac aatccaactt tggcatatgt atatgaaatc
     2521 aaaacaattc actactcatt cataatattg ccgaacttta ccaaattaaa aataaacatg
     2581 aaatagtcaa caactcattc gttatgtaac gttttttttt taaatgcatt aatatataat
     2641 gattgttttt attttggaag ttgaaataaa ttagggcaca accagtttcc actattctcc
     2701 tttatcttta cggaaatagc atgaataata ccgaaaaggc cgttttaaaa ataaaaattc
     2761 gagaaaattc aaacgcaaat caatggaaac ttcgagatca ttcggtgaag ttcgagacga
     2821 attccccgtt tcatatataa gcgcggtctc gcggcgcgtt gtcagtgaat ttgaatcaag
     2881 cgaagcgaaa gcaacaaagc taagcaagcg actcaaagca aattttcaaa gcgactttaa
     2941 caaagttgag tgaattattt actagtgaat tcaaagtcgt cgataaataa gtttcccatt
     3001 tttattcccc agccaggaag ttagtttcaa tagttttgta atttcaacga aactcatttg
     3061 atttcgtact aattttccac atctctattt tctgcccgca gaataatcca aaatgcagat
     3121 cggggatccc accccaccca agaagaagcg caaggtggag gacgatcccg tcgttttaca
     3181 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     3241 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     3301 gtcgactcta gaggatcccc gggatccacc ggtcgccacc atggataaga aatactcaat
     3361 aggcttagat atcggcacaa atagcgtcgg atgggcggtg atcactgatg aatataaggt
     3421 tccgtctaaa aagttcaagg ttctgggaaa tacagaccgc cacagtatca aaaaaaatct
     3481 tataggggct cttttatttg acagtggaga gacagcggaa gcgactcgtc tcaaacggac
     3541 agctcgtaga aggtatacac gtcggaagaa tcgtatttgt tatctacagg agattttttc
     3601 aaatgagatg gcgaaagtag atgatagttt ctttcatcga cttgaagagt cttttttggt
     3661 ggaagaagac aagaagcatg aacgtcatcc tatttttgga aatatagtag atgaagttgc
     3721 ttatcatgag aaatatccaa ctatctatca tctgcgaaaa aaattggtag attctactga
     3781 taaagcggat ttgcgcttaa tctatttggc cttagcgcat atgattaagt ttcgtggtca
     3841 ttttttgatt gagggagatt taaatcctga taatagtgat gtggacaaac tatttatcca
     3901 gttggtacaa acctacaatc aattatttga agaaaaccct attaacgcaa gtggagtaga
     3961 tgctaaagcg attctttctg cacgattgag taaatcaaga cgattagaaa atctcattgc
     4021 tcagctcccc ggtgagaaga aaaatggctt atttgggaat ctcattgctt tgtcattggg
     4081 tttgacccct aattttaaat caaattttga tttggcagaa gatgctaaat tacagctttc
     4141 aaaagatact tacgatgatg atttagataa tttattggcg caaattggag atcaatatgc
     4201 tgatttgttt ttggcagcta agaatttatc agatgctatt ttactttcag atatcctaag
     4261 agtaaatact gaaataacta aggctcccct atcagcttca atgattaaac gctacgatga
     4321 acatcatcaa gacttgactc ttttaaaagc tttagttcga caacaacttc cagaaaagta
     4381 taaagaaatc ttttttgatc aatcaaaaaa cggatatgca ggttatattg atgggggagc
     4441 tagccaagaa gaattttata aatttatcaa accaatttta gaaaaaatgg atggtactga
     4501 ggaattattg gtgaaactaa atcgtgaaga tttgctgcgc aagcaacgga cctttgacaa
     4561 cggctctatt ccccatcaaa ttcacttggg tgagctgcat gctattttga gaagacaaga
     4621 agacttttat ccatttttaa aagacaatcg tgagaagatt gaaaaaatct tgacttttcg
     4681 aattccttat tatgttggtc cattggcgcg tggcaatagt cgttttgcat ggatgactcg
     4741 gaagtctgaa gaaacaatta ccccatggaa ttttgaagaa gttgtcgata aaggtgcttc
     4801 agctcaatca tttattgaac gcatgacaaa ctttgataaa aatcttccaa atgaaaaagt
     4861 actaccaaaa catagtttgc tttatgagta ttttacggtt tataacgaat tgacaaaggt
     4921 caaatatgtt actgaaggaa tgcgaaaacc agcatttctt tcaggtgaac agaagaaagc
     4981 cattgttgat ttactcttca aaacaaatcg aaaagtaacc gttaagcaat taaaagaaga
     5041 ttatttcaaa aaaatagaat gttttgatag tgttgaaatt tcaggagttg aagatagatt
     5101 taatgcttca ttaggtacct accatgattt gctaaaaatt attaaagata aagatttttt
     5161 ggataatgaa gaaaatgaag atatcttaga ggatattgtt ttaacattga ccttatttga
     5221 agatagggag atgattgagg aaagacttaa aacatatgct cacctctttg atgataaggt
     5281 gatgaaacag cttaaacgtc gccgttatac tggttgggga cgtttgtctc gaaaattgat
     5341 taatggtatt agggataagc aatctggcaa aacaatatta gattttttga aatcagatgg
     5401 ttttgccaat cgcaatttta tgcagctgat ccatgatgat agtttgacat ttaaagaaga
     5461 cattcaaaaa gcacaagtgt ctggacaagg cgatagttta catgaacata ttgcaaattt
     5521 agctggtagc cctgctatta aaaaaggtat tttacagact gtaaaagttg ttgatgaatt
     5581 ggtcaaagta atggggcggc ataagccaga aaatatcgtt attgaaatgg cacgtgaaaa
     5641 tcagacaact caaaagggcc agaaaaattc gcgagagcgt atgaaacgaa tcgaagaagg
     5701 tatcaaagaa ttaggaagtc agattcttaa agagcatcct gttgaaaata ctcaattgca
     5761 aaatgaaaag ctctatctct attatctcca aaatggaaga gacatgtatg tggaccaaga
     5821 attagatatt aatcgtttaa gtgattatga tgtcgatcac attgttccac aaagtttcct
     5881 taaagacgat tcaatagaca ataaggtctt aacgcgttct gataaaaatc gtggtaaatc
     5941 ggataacgtt ccaagtgaag aagtagtcaa aaagatgaaa aactattgga gacaacttct
     6001 aaacgccaag ttaatcactc aacgtaagtt tgataattta acgaaagctg aacgtggagg
     6061 tttgagtgaa cttgataaag ctggttttat caaacgccaa ttggttgaaa ctcgccaaat
     6121 cactaagcat gtggcacaaa ttttggatag tcgcatgaat actaaatacg atgaaaatga
     6181 taaacttatt cgagaggtta aagtgattac cttaaaatct aaattagttt ctgacttccg
     6241 aaaagatttc caattctata aagtacgtga gattaacaat taccatcatg cccatgatgc
     6301 gtatctaaat gccgtcgttg gaactgcttt gattaagaaa tatccaaaac ttgaatcgga
     6361 gtttgtctat ggtgattata aagtttatga tgttcgtaaa atgattgcta agtctgagca
     6421 agaaataggc aaagcaaccg caaaatattt cttttactct aatatcatga acttcttcaa
     6481 aacagaaatt acacttgcaa atggagagat tcgcaaacgc cctctaatcg aaactaatgg
     6541 ggaaactgga gaaattgtct gggataaagg gcgagatttt gccacagtgc gcaaagtatt
     6601 gtccatgccc caagtcaata ttgtcaagaa aacagaagta cagacaggcg gattctccaa
     6661 ggagtcaatt ttaccaaaaa gaaattcgga caagcttatt gctcgtaaaa aagactggga
     6721 tccaaaaaaa tatggtggtt ttgatagtcc aacggtagct tattcagtcc tagtggttgc
     6781 taaggtggaa aaagggaaat cgaagaagtt aaaatccgtt aaagagttac tagggatcac
     6841 aattatggaa agaagttcct ttgaaaaaaa tccgattgac tttttagaag ctaaaggata
     6901 taaggaagtt aaaaaagact taatcattaa actacctaaa tatagtcttt ttgagttaga
     6961 aaacggtcgt aaacggatgc tggctagtgc cggagaatta caaaaaggaa atgagctggc
     7021 tctgccaagc aaatatgtga attttttata tttagctagt cattatgaaa agttgaaggg
     7081 tagtccagaa gataacgaac aaaaacaatt gtttgtggag cagcataagc attatttaga
     7141 tgagattatt gagcaaatca gtgaattttc taagcgtgtt attttagcag atgccaattt
     7201 agataaagtt cttagtgcat ataacaaaca tagagacaaa ccaatacgtg aacaagcaga
     7261 aaatattatt catttattta cgttgacgaa tcttggagct cccgctgctt ttaaatattt
     7321 tgatacaaca attgatcgta aacgatatac gtctacaaaa gaagttttag atgccactct
     7381 tatccatcaa tccatcactg gtctttatga aacacgcatt gatttgagtc agctaggagg
     7441 tgacggttct cccaagaaga agaggaaagt ctcgagcggt ggagctgcag gatgagcggc
     7501 cgcctgcagc tggcgccatc gatacgcgta cgtcgcgacc gcggacatgt acagagctcg
     7561 agatgatcgg tttgtgttcc tacttagtta attgtgatag tttatttttg taaataatag
     7621 tacgtaaagt tactttagtt ataataaagt agatagtgac tgcattaagt ttaaagactg
     7681 atttttgtag attatttaaa aatttttaat aaattattta atttaaagat aaaaagttgt
     7741 ttcattttaa aataataaaa aacttaccgc attagtcgtg gccttggcag tattaagcgc
     7801 aaacgggctc tccttcgtat gcccatcagg aatatagaaa aaaacaactg gctgctcact
     7861 agtggccacg tgggccgtgc accttaagct tggcgcgcca aaaaagcacc gactcggtgc
     7921 cactttttca agttgataac ggactagcct tattttaact tgctatttct agctctaaaa
     7981 ctatacgaag ttattacgaa cgaacggtgc ttctacctgc cttgtatatg tagttagcgg
     8041 caaccgatac gtagtagtga gatataagag atttgaaatt tcccgcttta ttttgattgg
     8101 tcggttacaa tcgatcgcaa aacacaaacg ttcgtaatga ttcgtatatt tgtatgtaat
     8161 ggcgcttatt taaaattttt attatttagt tgaagaaata ttaatttgtc gtaatgaaaa
     8221 cagttttgaa acaaaaatag atattcattt gttcataagg taactgataa atctgcgtgt
     8281 tgtgttttta aataatcctt gaacagtgac tcaatctcta gtacttgaag cacttatcac
     8341 caggtcacaa gtaataataa taacacccgg cgcgccgtac gcgtatcgat aagctttaag
     8401 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     8461 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     8521 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     8581 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatctagagt cgcggccgcc
     8641 ttaacgaaat aattaattgt ttccattcga cacgttattg gtgctattcg ccgaccccca
     8701 attgcacaaa tgacttctca tagacgtaga caaggggggg atgtaggacc gtggtatcaa
     8761 aaacgtcgat aaattgaaaa ggtccacgaa aactttatac cgatcgctga aaaacaaaat
     8821 caaacattgc aaaaaccgtg ttgaaatcaa tttcacctta acgttgacct taaatactga
     8881 taccccgaag agccccctgt tcctaactgc gaagaaccaa tcattctttg caccatcaaa
     8941 acatgattat ctgaaattat aaaatttaaa caaaaacaat tgacgtacgg atttactaac
     9001 accgccattt cgtaatcagc gagtctatgt ccatcaaaag tgttagaatt tggtggggct
     9061 gactgaacct gggttcgtcc ctgtaaaagg tgatcatgat agccccttgc agggctttgt
     9121 gtgagaagcg cctctcgccg cgcgacatta gagcctcatg gacctcaact ttaaaaatcg
     9181 actcgtaaac ctcgcgtctc ttctccaaat cgttcaattt gtaacgcttt aaagtctccg
     9241 cctcctcttt ctccgccagt tctttttgct cggttaaaag cttctcgacc gctttttggt
     9301 acttccccca gaaattaaac ccctcgagct ccagccccgg cgttcgctcc agccaccgct
     9361 ggaccaaatc ggtcaaagac ggctctttct ccgacttggc gatttgctcg agcgcttttt
     9421 cgtcgtttcc gaacactttc gagtaattct ggttgtattt gacgcggttt tcctggcgca
     9481 ccccgagctt attttccagg agcctgaact gcaggctctg gaacccggaa gcgggccgca
     9541 gataacaccg gaagtccatg aaatcgagcg gtgtcatcgt ttccagaatc attacttggt
     9601 ccaccaaaac ctacaattgc gacagtttta tctcgtgacc gcaatttgtt gcgaaattgt
     9661 gaaccttgag gatcaaaaca acacgattga ggcgtttgag gatttccaaa gtttgcgact
     9721 cttctaaaac gtcgctgaaa atgttgcgga tcgagtccag ctcgtagatt atttgtttga
     9781 accataactc atacgctgga attacttgtt atagttaatt acggttaagt gggtttttta
     9841 ccttgatggg tcacaataaa taagtgctcg tcgtgcacgg gttggttatt ttgctcgctg
     9901 agtagtcttt gggcttctaa aatcttgtcc aacatcaagt attcaccgta aagcatccca
     9961 cattcttcgc tcagttggtc gccttcttgg gcttcactta atttacgatt aattgtaaat
    10021 ggaataatta aattaaatca tacttacgag ggtctcagtg ggcaactcat tacgaccgcg
    10081 gtaccgtcga ctctagcggt accccgattg tttagcttgt tcagctgcgc ttgtttattt
    10141 gcttagcttt cgcttagcga cgtgttcact ttgcttgttt gaattgaatt gtcgctccgt
    10201 agacgaagcg cctctattta tactccggcg gtcgagggtt cgaaatcgat aagcttggat
    10261 cctaattgaa ttagctctaa ttgaattagt ctctaattga attagatccc cgggcgagct
    10321 cgaattaacc attgtgggaa ccgtgcgatc aaacaaacgc gagataccgg aagtactgaa
    10381 aaacagtcgc tccaggccag tgggaacatc gatgttttgt tttgacggac cccttactct
    10441 cgtctcatat aaaccgaagc cagctaagat ggtatactta ttatcatctt gtgatgagga
    10501 tgcttctatc aacgaaagta ccggtaaacc gcaaatggtt atgtattata atcaaactaa
    10561 aggcggagtg gacacgctag accaaatgtg ttctgtgatg acctgcagta ggaagacgaa
    10621 taggtggcct atggcattat tgtacggaat gataaacatt gcctgcataa attcttttat
    10681 tatatacagc cataatgtca gtagcaaggg agaaaaggtt caaagtcgca aaaaatttat
    10741 gagaaacctt tacatgagcc tgacgtcatc gtttatgcgt aagcgtttag aagctcctac
    10801 tttgaagaga tatttgcgcg ataatatctc taatattttg ccaaatgaag tgcctggtac
    10861 atcagatgac agtactgaag agccagtaat gaaaaaacgt acttactgta cttactgccc
    10921 ctctaaaata aggcgaaagg caaatgcatc gtgcaaaaaa tgcaaaaaag ttatttgtcg
    10981 agagcataat attgatatgt gccaaagttg tttctgactg actaataagt ataatttgtt
    11041 tctattatgt ataagttaag ctaattactt attttataat acaacatgac tgtttttaaa
    11101 gtacaaaata agtttatttt tgtaaaagag agaatgttta aaagttttgt tactttatag
    11161 aagaaatttt gagtttttgt ttttttttaa taaataaata aacataaata aattgtttgt
    11221 tgaatttatt attagtatgt aagtgtaaat ataataaaac ttaatatcta ttcaaattaa
    11281 taaataaacc tcgatataca gaccgataaa acacatgcgt caattttacg catgattatc
    11341 tttaacgtac gtcacaatat gattatcttt ctagggttaa ataatagttt ctaatttttt
    11401 tattattcag cctgctgtcg tgaataccgt atatctcaac gctgtctgtg agattgtcgt
    11461 attctagcct ttttagtttt tcgctcatcg acttgatatt gtccgacaca ttttcgtcga
    11521 tttgcgtttt gatcaaagac ttgagcagag acacgttaat caactgttca aattgatcca
    11581 tattaacgat atcaacccga tgcgtatatg gtgcgtaaaa tatatttttt aaccctctta
    11641 tactttgcac tctgcgttaa tacgcgttcg tgtacagacg taatcatgtt ttcttttttg
    11701 gataaaactc ctactgagtt tgacctcata ttagaccctc acaagttgca aaacgtggca
    11761 ttttttacca atgaagaatt taaagttatt ttaaaaaatt tcatcacaga tttaaagaag
    11821 aaccaaaaat taaattattt caacagttta atcgaccagt taatcaacgt gtacacagac
    11881 gcgtcggcaa aaaacacgca gcccgacgtg ttggctaaaa ttattaaatc aacttgtgtt
    11941 atagtcacgg atttgccgtc caacgtgttc ctcaaaaagt tgaagaccaa caagtttacg
    12001 gacactatta attatttgat tttgccccac ttcattttgt gggatcacaa ttttgttata
    12061 tttttaaaca aagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
    12121 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
    12181 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
    12241 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
    12301 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
    12361 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
    12421 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
    12481 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
    12541 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
    12601 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
    12661 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
    12721 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
    12781 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
    12841 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
    12901 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
    12961 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
    13021 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
    13081 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
    13141 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
    13201 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
    13261 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
    13321 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
    13381 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
    13441 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
    13501 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
    13561 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
    13621 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
    13681 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
    13741 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
    13801 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
    13861 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
    13921 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
    13981 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
    14041 cacagcccag cttggagcga acgacctaca ccgaact
//
