LOCUS       sequence-229414-       16006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targets 2 genes and expresses zebrafish mitfa specifically in 
            zebrafish melanocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16006)
  AUTHORS   Ablain J, Xu M, Rothschild H, Jordan RC, Mito JK, Daniels BH, Bell 
            CF, Joseph NM, Wu H, Bastian BC, Zon LI, Yeh I
  TITLE     Human tumor genomics and zebrafish modeling identify SPRED1 loss as 
            a driver of mucosal melanoma.
  JOURNAL   Science. 2018 Nov 1. pii: science.aau6509. doi: 
            10.1126/science.aau6509.
  PUBMED    30385465
REFERENCE   2  (bases 1 to 16006)
  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: "Science. 
            2018 Nov 1. pii: science.aau6509. doi: 10.1126/science.aau6509."
COMMENT     Sequence Label: MiniCoopR 2xU6:gRNA, mitfa:Cas9
FEATURES             Location/Qualifiers
     source          1..16006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..124
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(125..193,194..985)
                     /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(343..362)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1156..1744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1645..1664
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1898..1915
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2032..2053
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2068..2073,2074..2091,2092..2098)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2106..2122
                     /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     2111..2133
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2130..2146
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2167..2185
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2222..2238
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2222..2238
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    join(2272..2283,2284..2471)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     polyA_signal    7262..7396
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_RNA        7795..7870
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        8273..8348
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     8361..8377
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8385..8405
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    8468..8602
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    8604..8628
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        8657..8675
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(8729..8749)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8765..12865)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes MGAS15252 Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFGSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLADSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQIYNQLFEENPINASRVDAKAILSARLSKSRRL
                     ENLIAQLPGEKRNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNSEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGAYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRGMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGHSLHEQIANLAGSPAIKKGILQTVKIVDELVKVMGH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFIKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             8844..8852
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(12872..12892)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      12901..12910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(12945..12964)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(12946..12964)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(12974..12990)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(12992..13016)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(14890..14910)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(14930..14946)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    join(15319..15481,15482..15493)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     15842..15863
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
       61 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      121 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      181 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
      241 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
      301 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      361 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      421 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      481 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
      541 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
      601 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
      661 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
      721 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
      781 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
      841 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
      901 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
      961 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1021 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1081 catgaccaaa atgccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1141 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1201 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1261 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1321 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1381 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1441 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1501 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1561 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1621 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1681 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     1741 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     1801 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     1861 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     1921 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     1981 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     2041 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     2101 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     2161 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctccaccgcg gtggcggccg
     2221 ctctagaact agtggatctg ctgggcttgc tgaaggtagg gggtcaagaa ccagaggtgt
     2281 aaagtacttg agtaatttta cttgattact gtacttaagt attatttttg gggattttta
     2341 ctttacttga gtacaattaa aaatcaatac ttttactttt acttaattac atttttttag
     2401 aaaaaaaagt actttttact ccttacaatt ttatttacag tcaaaaagta cttatttttt
     2461 ggagatcact tcattctatt ttcccttgct attaccaaac caattgaatt gcgctgatgc
     2521 ccagtttaat ttaaatgtta tttattctgc ctatgaaaat cgttttcaca ttatatgaaa
     2581 ttggtcagac atgttcattg gtcctttgga agtgacgtca tgtcacatct attaccacaa
     2641 tgcacagcac cttgacctgg aaattaggga aattataaca gtcaatcagt ggaagaaaat
     2701 ggaggaagta tgtgattcat cagcagctgc gagcagcaca gtccaaaatc agccacagga
     2761 tcaagagcac ccgtggccgt atcttcgcag atctcgaata agagattgct acggagagat
     2821 attctgctga ttaggagctt gtctatgcta atttggtttt gttcaattca cctatgtcgc
     2881 aagtttttgg actgtaggag aaaaccggag aacccagggg atatccacat gagcacgagg
     2941 acaacatgta aactccacac agaaccatgg actggcctgt tagagacttg gaccagtgac
     3001 gttcttgctt tgaggcaaca gtgctaacaa ctgggctgct gtgccgctct ttcgaggaaa
     3061 tgaaaggaac tctggcggcc tatgatgagt taggaattgt ctgattggtg aatcatacat
     3121 aggctaatgt gggaccagct gtagtcagtc ataagcacgt gattctctcg aaattagttt
     3181 ataaataaac ttcactaaac tccaaccgtt cattctgttt tagatgttgc ctttaaactt
     3241 gccttaagat gttaataaag ttgctttttg gttgttgaca aagctaggca taggacgata
     3301 accgttttca aggtataccc cggtttgaaa aagtcaaggg tttaaaacca ccaatatttt
     3361 ctgctatacc gttcctaagg tatatttaag attttttatt tattttttat ttttttgttt
     3421 tttaggacaa cagcattttc agcagaaaag agatacaaaa atgcttttta aattgtaaag
     3481 aaatctgttt ttgaaactaa tgaagacagc agaagtcgat cattcattga aattattaag
     3541 cttgacatgt tttctgttcc aaaatatttt ttcaatgttt ctcaaaataa aatatattat
     3601 gtttaaagtt tttgtttttt actaagacat taaaaaaaat aatatatttt agagtggcaa
     3661 tcacaatacc atcaaactgt gatattttta tcaaaggtta tcataccatc agaatcttat
     3721 accccatgcc tagacacagc tgttctctct atctctctat gttaatgcag tttccactcc
     3781 attgacgaga tggttctctg taccacatac tcatccccat tttacaatag ctacatttgc
     3841 tcaagatgta gtcaactcaa acaatgcaga gacaacaaac tgttgactta tcttttctat
     3901 cgtaattcag cccttttggg aaatgagcat caacacacaa tttagagtgg aagttgggtt
     3961 aaggcagact cattttttac acattacaac cagtgattat gtcaattatt atttaaaaaa
     4021 aattcagtac agtatatgca catgctactt tgtttatgat tgttagcttt tttaactaca
     4081 gttcatttct atgctttctg tgcaaatgca caacagttac tgatttactc tgatgataat
     4141 tgtgccagta catccaacaa caacaaaaaa aaaaaaaaga cttacaatca agccccaaag
     4201 tgttcatacg tctggcaaat tctgacttaa agttactttt attcaaccag ctagtttttt
     4261 tttgactgta aatgacacgt aagataataa gatgatgtac atgaggcatc attgtagaaa
     4321 aaaaaatgaa tatgaatctg aatatgaact gtacgtctgt gattttgttt gtgcgcactt
     4381 acatacgtgt gcacaatatt tcaactctta agaaaaaaat gcaattacca aaggcccatc
     4441 agacaacaaa gttctctttc ctatcttttg ctcttcaaca tttcagagat aactccatct
     4501 taaactgtct tgtttttaga ccaacagtgc tagtgattgt atgccggatg ctgagaacaa
     4561 acaatgtttt atgcaggatg agtggaacta aagatgaaac gttagttagc cgtaagccag
     4621 tttcagttta gacactagta acccatcgtc tcaaagaggc ttttgtcgaa tcggaccggt
     4681 gacgtcaaat gaactttata aaggcataaa ttcatcgttt gggtaaaaaa aacaatatga
     4741 gaagagcaat cccctcccac ccccttaaat acattttgtg tcttttcttt ggcattgaca
     4801 tcttcagctg gccaagacga ctggtcagtt cttgcacatc acctaaattc acaaagggaa
     4861 tacttttttg aagctaacac atagttgaac atgttggaga tgctcgagta cagtcactac
     4921 caggttcaga cccacctgga aaccccctcg aagtaccaca tccagcagag ccagaggcag
     4981 caagtgaagc actacctgtc cagtgcactg ggagccaagc tgagcccaca ggccagcaca
     5041 gggcccggcc ccagccagcc cgccgagcac ggcatgaccc cgggacccgg agccagcgct
     5101 cccaacagcc ctatggccct tctcaccctc aactgtgaga aagagatgga cgatgtaatt
     5161 gaagacatta taagtttgga atcaagctac agtgatgaca ttcttgggtt catggatgca
     5221 ggacttcaaa tgacaaacac gattccagtt tcagcaaacc tgctggacat gtacagcaat
     5281 catgctcttc ctccagctgg agtttccatc agtaactcct gcccctcaag cctgccggcc
     5341 gtcaaaaggg aattatccgt tactccatct ccgggcatga tgcacataat ggacaaagct
     5401 ggaccatgtg gcaagtttga ctcttatcaa agacctgatg gctttccagt agaagcagaa
     5461 gtcagagccc tggcaaaaga gagacaaaag aaggacaacc acaacctcat tgaacgaaga
     5521 aggcggttta atatcaatga cagaattaag gagctgggga ctttaattcc caagtcaaat
     5581 gatccagaca tgaggtggaa taaaggcacc atcctgaaag catcagtgga ttacattagg
     5641 aaattgcaga aagagcagca gaaagcaaaa gagctggaaa acagacagaa gagactagaa
     5701 cacgcaaaca gacatctcct gctccgtatt caggaacttg aaatgcaagc ccgtgctcat
     5761 ggactcaccg ttgtagcttc ttccagcctt tattccgctg agttagtggc tcgagcaatc
     5821 aaacaagagc ctgggatggg ggactgcaca tccaatttgt acccacacct tcccagtcct
     5881 gacatgtccc gtcctaccac tctggacctc aacaacggca ccatcagcta caatgacagt
     5941 cccacggagg acggcgagcc gggggtctac gacagcccaa acaaggcctc caccaagctg
     6001 gaggacatgc ttatggacaa caccctgtct ccggtgggca gcagcgaccc gctgttgtcc
     6061 tccggatccc cagtaccttc caacagtagc ggcagcagca gtatggacga acatgacaat
     6121 ggctgttagc aatgccaact aaatttcatg aaccaaagtt agacaattag atttgtgtgt
     6181 gtgtgtgtct tgtaagtacc agtgttacat ccagatcatg aagagcgcaa tttcgtaaag
     6241 tattaatctt gtatttcaga atatttagtg tgcttgtgta tatatttttt catttatcat
     6301 gtcagagtct agtcaagaat gaaatcgtgt aaacaattag ttgatttttg aaggattctc
     6361 gcggatgata taaagagtag cttgaataaa aactgttgag gtgtctttta tttgcaggct
     6421 tggcttattg ttgtcctcaa atgcagctcc atttcttggg gtgtacatct tgcaagggtg
     6481 gaggacatgt ctaggcatgt acaactggat tcactttcat tcagtcattc attttctttt
     6541 tggcttagcc cctttattaa tcaggggttg ccacagttga atgaactgcc aacttatgcc
     6601 aacaaacaaa tccagcggat gcccttgcag ctgcaaccca tcactgggaa acacccacca
     6661 cagtaccatt cactttagac aatttagctt acccaattca cctatagcgc atgtctttgg
     6721 gctgtaggtg aaactggagc acccggagga aacccacacg aacatgggga gaacatgcaa
     6781 actccacaca gaaatgaccc aaccaagtct cgaaccagcg accttcttgc tgtaaggcga
     6841 caatgctacc cactgcacca ccacgatgcc ctggaaaaac tttttacatt gttatttaac
     6901 attaagtcca aaagtatttc tgcagtattt ctgaaatttg ttttattttc tagtggaaga
     6961 gctgtgatgc aagttctcca taataattag aaaaaaaatg cccaaattgc tttgatttta
     7021 gggatagttc acccaaaaat gaattttttt ttattaatta atcatcctca tgtcgttcca
     7081 attccctgag acctttgttc atctttaaaa cacaaaaaat gatattttgg atacaatctg
     7141 agatctgatc tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt
     7201 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     7261 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     7321 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     7381 ttatcatgtc tggatcatca tcgatgcgtc ttttgttctg gtcatcaagg aggggggaat
     7441 gttctgcgca tgcctgtggg ggagggagaa ggacacgtca ctgaaaacgt ccctgcatca
     7501 caccgagaca cccaatcact caagccgaga ccagataatt ttgcatatgc tttaaagttt
     7561 gaaaaatacc acggtaaacc ctcacacaaa ctctggattt gagatctttc aggttttatc
     7621 agtttgcagg tttatgtcac catgatatag ggtcagactt gatctaaggg agctgaataa
     7681 gtggtttagt cactcaccac ctcccaaaaa catacccaga agtccctggt atatatagct
     7741 ctccctccag ctcttggttc ggctagcact cctcagtgac gaggagtcag tagcgtttta
     7801 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
     7861 gagtcggtgc ttttttggtc gacctcgagg cgtcttttgt tctggtcatc aaggaggggg
     7921 gaatgttctg cgcatgcctg tgggggaggg agaaggacac gtcactgaaa acgtccctgc
     7981 atcacaccga gacacccaat cactcaagcc gagaccagat aattttgcat atgctttaaa
     8041 gtttgaaaaa taccacggta aaccctcaca caaactctgg atttgagatc tttcaggttt
     8101 tatcagtttg caggtttatg tcaccatgat atagggtcag acttgatcta agggagctga
     8161 ataagtggtt tagtcactca ccacctccca aaaacatacc cagaagtccc tggtatatat
     8221 agctctccct ccagctcttg gttcgcatgc aggtgtgact gcacctgcat aggttttaga
     8281 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     8341 gtcggtgctt ttttggtacc gtaaaacgac ggccagtgaa ttatcaacta tgtataataa
     8401 agttggaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     8461 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     8521 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     8581 tgtatcttat catgtctgga tcccccactt tgtacaagaa agctgggtgc tatgaccatg
     8641 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctccaccgcg
     8701 gtggcggccg ctctagattc cgcggtcaca cctttctctt cttcttagga gatctcaccg
     8761 gtgagtcgcc acccagctga gacaggtcaa tccttgtttc gtacaggccg gtaatactct
     8821 gatggatcag tgtagcgtcc agcacctcct tagtggaggt gtacctcttt ctatcgattg
     8881 tagtgtcgaa gtacttgaaa gctgctggtg cgcccaggtt tgtcagagtg aacagatgaa
     8941 tgatattttc ggcctgctcc ctgatgggct tatctctgtg tttgttgtaa gcagacagga
     9001 ccttatccag atttgcgtcg gccaggatca ctcttttaga gaactcgcta atctgttcaa
     9061 tgatctcgtc cagataatgc ttgtgctgct ccacaaacag ctgtttctgt tcgttatcct
     9121 ctggagatcc tttcagcttc tcgtagtggc ttgccagata caggaagttc acgtacttag
     9181 aaggcagggc cagttcattg cctttctgca gctctccagc gcttgccagc atgcgtttcc
     9241 tgccgttttc cagctcaaac agactgtact tgggcagttt aatgatcagg tctttcttca
     9301 cttccttata acctttggcc tccagaaaat cgattgggtt cttctcgaaa gagcttctct
     9361 ccataattgt gatacccagc agctccttga cagatttcag tttcttgctt ttgcccttct
     9421 ccaccttagc gaccaccagg acggagtatg ccacagtggg agagtcaaag cctccatact
     9481 tcttagggtc ccagtctttc tttctagcga tcagcttatc agagttgcgt tttggcagaa
     9541 tactttcttt tgagaaaccg cctgtctgca cctcagtttt cttgacgata ttcacttgag
     9601 gcatgctcag gaccttcctc acggttgcaa aatctctgcc tttgtcccac acgatttcgc
     9661 ctgtctctcc gtttgtctca atcaggggcc ttttgcggat ttctccattg gccagggtga
     9721 tctctgtctt aaagaaattc atgatgtttg agtaaaagaa atacttggca gtagcttttc
     9781 caatttcctg ctcggacttg gcgatcattt ttctgacatc gtacacctta tagtctccgt
     9841 acacgaattc gctctccagt tttgggtatt tcttgatcag tgcggtaccg accacggcgt
     9901 tcaggtatgc gtcatgggca tggtgataat tgttaatctc gcggacctta taaaactgga
     9961 aatctttcct aaagtcgctg accagtttac tcttcagggt aatgaccttc acttctctga
    10021 tcagtttgtc attctcatcg tactttgtgt tcatgcgact atccagaatc tgtgcgacgt
    10081 gtttagtgat ctggcgtgtt tccaccagct gcctcttgat gaatccagct ttgtccagtt
    10141 ctgacagtcc accgcgctct gcttttgtca ggttatcaaa cttcctctga gtgatcagtt
    10201 tggcattcag cagctgtctc cagtagttct tcattttctt gaccacttcc tcactaggga
    10261 cattatctga ttttcctctg tttttgtcgg acctggtcag caccttgttg tcaatgctgt
    10321 catctttgat gaaactctgg gggacgatat gatccacgtc gtaatcagac agcctgttaa
    10381 tatccagctc ctggtccacg tacatatctc ttccattctg cagatagtac agatacagct
    10441 tctcattctg cagctgtgtg ttttccactg ggtgctcttt caggatctgg ctacccagct
    10501 ccttaatgcc ttcctcgatt cttttcatcc tctctctact gttcttctga cccttctggg
    10561 ttgtctgatt ttcgcgagcc atctcaatca cgatgttttc gggcttatgg cccatgactt
    10621 tcaccagctc atcgacgatt ttcactgtct gcagaatgcc tttcttgatg gctgggcttc
    10681 cagccaggtt tgcaatctgc tcatgcaggg agtggccctg tccagacacc tgagccttct
    10741 ggatgtcctc tttgaaggtc agactgtcat cgtgaatcag ctgcataaaa ttgcggttgg
    10801 cgaagccatc acttttcaga aagtccagga ttgtctttcc tgactgttta tccctaatgc
    10861 cgttgatcag cttcctgctc agtctacccc agccagtgta gcgccttctc ttcagctgtt
    10921 tcatcacctt gtcatcgaac agatgagcat aggttttcag tctttcctcg atcattcctc
    10981 tgtcctcaaa cagggtcagt gtcagcacaa tgtcttccag gatatcctca ttttcctcgt
    11041 tgtccaggaa gtctttatcc ttaatgatct tcagcagatc gtggtaagcg cccaggcttg
    11101 cgttaaatct gtcttcgact ccgctgatct ccacggaatc gaagcattca attttcttga
    11161 agtagtcctc cttcagctgc ttgacagtca ctttcctgtt ggttttaaac agcagatcca
    11221 cgattgcttt cttctgctct cctgacagga aggctggctt tctcatgccc tcggtgacgt
    11281 atttcacctt tgtcagttcg ttatagacag tgaaatactc gtacagcagg gagtgcttag
    11341 gcagcacttt ctcattgggc aggtttttgt caaaattagt cattctctcg atgaaggact
    11401 gtgcagaggc gcccttatcg accacttcct cgaagttcca aggagtaatg gtttcctcgg
    11461 atttgcgagt catccaggca aatctgctgt ttccgcgagc cagtggaccc acatagtaag
    11521 ggattctgaa ggtcagaatt ttttcgatct tctcgcggtt atctttcaga aaggggtaga
    11581 agtcttcctg gcgcctcagg atagcgtgca gctctcccag atgaatctgg tgtgggatgg
    11641 aaccgttatc aaatgtgcgc tgcttcctca gcaggtcctc tctattcagt ttcaccagca
    11701 gttcctcagt tccgtccatc ttctccagaa taggtttgat aaacttgtag aattcctcct
    11761 gtgatgcgcc tccgtcgata taaccggcgt agccgttctt ggactgatca aagaaaattt
    11821 ccttgtactt ctctggcagc tgctgcctga ccagagcctt cagcagggtc agatcctgat
    11881 ggtgctcgtc atatctcttg atcatgctgg cactcagagg tgcttttgta atctcgctgt
    11941 tcactctcag aatatctgac agcaggattg cgtcggacag atttttggca gccaggaaca
    12001 ggtcagcgta ctgatctccg atctgtgcca gcaggttatc caggtcatcg tcatatgtgt
    12061 cctttgacag ctgcagtttg gcgtcctcag ccagatcaaa attgcttttg aagttaggag
    12121 tcaggcccag actcagagcg atcagatttc caaacagacc gttcctcttt tctccgggca
    12181 gctgtgcaat caggttctcc agtctgcgtg acttggacag cctggctgac aggatagctt
    12241 ttgcgtccac tctggaagca ttaatagggt tttcctcaaa cagctgattg taaatctgga
    12301 ccagctggat gaacagcttg tccacatcgg aattgtcggg gttcagatcg ccctcgatca
    12361 gaaaatgacc cctgaactta atcatgtggg ccagagccag atagatcagt ctcaggtcgg
    12421 ctttatcagt agagtcagcc agtttcttgc gcagatgata gatggttggg tatttttcgt
    12481 gataggcgac ctcgtccaca atgtttccaa agataggatg gcgctcgtgt ttcttatctt
    12541 cctcgaccag gaaactttcc tccagcctat gaaagaatga atcgtccacc ttagccattt
    12601 cgttagagaa aatctcctgc aggtaacaga ttctattctt ccttctggta tagcgccttc
    12661 ttgctgttct tttcaggcgt gttgcttcgg cagtctctcc agaaccgaac agcagggctc
    12721 caatcagatt tttcttgatg ctgtgcctgt cggtgttacc cagcacctta aatttcttgc
    12781 tagggacctt gtagtcgtct gtaatcactg cccaacccac ggagttagtt ccaatatcca
    12841 ggccgatgct atacttctta tccatgcttc ccacctttct cttcttctta ggtggagaag
    12901 ccatggtggc gtcgacctcg agggggggcc cggtacccaa ttcgccctat agtgagtcgt
    12961 attacgcgcg ctcactggcc gtcgttttac aagcctgctt ttttgtacaa acttgcgttc
    13021 aactatgtgt tagcttcaaa aaagtattcc ctttgtgaat ttaggtgatg tgcaagaact
    13081 gaccagtcgt cttggccagc tgaagatgtc aatgccaaag aaaagacaca aaatgtattt
    13141 aagggggtgg gaggggattg ctcttctcat attgtttttt ttacccaaac gatgaattta
    13201 tgcctttata aagttcattt gacgtcaccg gtccgattcg acaaaagcct ctttgagacg
    13261 atgggttact agtgtctaaa ctgaaactgg cttacggcta actaacgttt catctttagt
    13321 tccactcatc ctgcataaaa cattgtttgt tctcagcatc cggcatacaa tcactagcac
    13381 tgttggtcta aaaacaagac agtttaagat ggagttatct ctgaaatgtt gaagagcaaa
    13441 agataggaaa gagaactttg ttgtctgatg ggcctttggt aattgcattt ttttcttaag
    13501 agttgaaata ttgtgcacac gtatgtaagt gcgcacaaac aaaatcacag acgtacagtt
    13561 catattcaga ttcatattca tttttttttc tacaatgatg cctcatgtac atcatcttat
    13621 tatcttacgt gtcatttaca gtcaaaaaaa aactagctgg ttgaataaaa gtaactttaa
    13681 gtcagaattt gccagacgta tgaacacttt ggggcttgat tgtaagtctt tttttttttt
    13741 ttgttgttgt tggatgtact ggcacaatta tcatcagagt aaatcagtaa ctgttgtgca
    13801 tttgcacaga aagcatagaa atgaactgta gttaaaaaag ctaacaatca taaacaaagt
    13861 agcatgtgca tatactgtac tgaatttttt ttaaataata attgacataa tcactggttg
    13921 taatgtgtaa aaaatgagtc tgccttaacc caacttccac tctaaattgt gtgttgatgc
    13981 tcatttccca aaagggctga attacgatag aaaagataag tcaacagttt gttgtctctg
    14041 cattgtttga gttgactaca tcttgagcaa atgtagctat tgtaaaatgg ggatgagtat
    14101 gtggtacaga gaaccatctc gtcaatggag tggaaactgc attaacatag agagatagag
    14161 agaacagctg tgtctaggca tggggtataa gattctgatg gtatgataac ctttgataaa
    14221 aatatcacag tttgatggta ttgtgattgc cactctaaaa tatattattt tttttaatgt
    14281 cttagtaaaa aacaaaaact ttaaacataa tatattttat tttgagaaac attgaaaaaa
    14341 tattttggaa cagaaaacat gtcaagctta ataatttcaa tgaatgatcg acttctgctg
    14401 tcttcattag tttcaaaaac agatttcttt acaatttaaa aagcattttt gtatctcttt
    14461 tctgctgaaa atgctgttgt cctaaaaaac aaaaaaataa aaaataaata aaaaatctta
    14521 aatatacctt aggaacggta tagcagaaaa tattggtggt tttaaaccct tgactttttc
    14581 aaaccggggt ataccttgaa aacggttatc gtcctatgcc tagctttgtc aacaaccaaa
    14641 aagcaacttt attaacatct taaggcaagt ttaaaggcaa catctaaaac agaatgaacg
    14701 gttggagttt agtgaagttt atttataaac taatttcgag agaatcacgt gcttatgact
    14761 gactacagct ggtcccacat tagcctatgt atgattcacc aatcagacaa ttcctaactc
    14821 atcataggcc gccagagttc ctttcatttc ctcgaaagag cggcacagca gcccagttgt
    14881 tagcactcac aacttttcta tacaaagttg atagcttggc gtaatcatgg tcatagctgt
    14941 ttcctgctcg agggcccatc tggcctgtgt ttcagacacc agggagtctc tgctcacgtt
    15001 tcctgctatt tgcagcctct ctatcaagac taatacacct cttcccgcat cggctgcctg
    15061 tgagaggctt ttcagcactg caggattgct tttcagcccc aaaagagcta ggcttgacac
    15121 taacaatttt gagaatcagc ttctactgaa gttaaatctg aggttttaca actttgagta
    15181 gcgtgtactg gcattagatt gtctgtctta tagtttgata attaaataca aacagttcta
    15241 aagcaggata aaaccttgta tgcatttcat ttaatgtttt ttgagattaa aagcttaaac
    15301 aagaatctct agttttcttt cttgctttta cttttacttc cttaatactc aagtacaatt
    15361 ttaatggagt acttttttac ttttactcaa gtaagattct agccagatac ttttactttt
    15421 aattgagtaa aattttccct aagtacttgt actttcactt gagtaaaatt tttgagtact
    15481 ttttacacct ctgtcaagaa ctcctggaca aacctctgac ctgtgtggaa cagagtggat
    15541 atgggtgtct gaacagatat tcacgtcttt tgcagatcag agggcatttc tggtgttgtg
    15601 caatgacctg ggtccaacag gtaggcgtac tcgtatccaa gagcattgtt ggacaggagg
    15661 tagtctccat tcctataaag aggaaggaat ggcaccatgt agtagccgtc attgtggcca
    15721 attggtgcat tggtacgtgg gtagatggac cggggaggct gatgagttct tagccagcgt
    15781 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
    15841 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
    15901 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
    15961 tttaacgcga attttaacaa aatattaacg cttacaattt aggtgg
//
