LOCUS       sequence-229410-       15514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targets spred1 and expresses zebrafish mitfa specifically in 
            zebrafish melanocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15514)
  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 15514)
  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 U6:gRNA spred1, mitfa:Cas9
FEATURES             Location/Qualifiers
     source          1..15514
                     /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        7781..7856
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     7869..7885
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7893..7913
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    7976..8110
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    8112..8136
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        8165..8183
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(8237..8257)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8273..12373)
                     /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             8352..8360
                     /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(12380..12400)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      12409..12418
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(12453..12472)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(12454..12472)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(12482..12498)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(12500..12524)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(14398..14418)
                     /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(14438..14454)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    join(14827..14989,14990..15001)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     15350..15371
                     /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 caggtgaagc 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 tttacagttt
     7561 gaaaaatacc acggtaaacc ctcacacaaa ctctggattt gagatctttc aggttttatc
     7621 agtttgcagg tttatgtcac catgatatag ggtcagactt gatctaaggg agctgaataa
     7681 gtggtttagt cactcaccac ctcccaaaaa catacccaga agtccctggt atatatagct
     7741 ctccctccag ctcttggttc ggcgtccgcc gggctctgga gttttagagc tagaaatagc
     7801 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     7861 ttggtaccgt aaaacgacgg ccagtgaatt atcaactatg tataataaag ttggaaaaaa
     7921 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     7981 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     8041 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     8101 tgtctggatc ccccactttg tacaagaaag ctgggtgcta tgaccatgat tacgccaagc
     8161 gcgcaattaa ccctcactaa agggaacaaa agctggagct ccaccgcggt ggcggccgct
     8221 ctagattccg cggtcacacc tttctcttct tcttaggaga tctcaccggt gagtcgccac
     8281 ccagctgaga caggtcaatc cttgtttcgt acaggccggt aatactctga tggatcagtg
     8341 tagcgtccag cacctcctta gtggaggtgt acctctttct atcgattgta gtgtcgaagt
     8401 acttgaaagc tgcaggtgcg cccaggtttg tcagagtgaa cagatgaatg atattttcgg
     8461 cctgctccct gatgggctta tctctgtgtt tgttgtaagc agacaggacc ttatccagat
     8521 ttgcgtcggc caggatcact cttttagaga actcgctaat ctgttcaatg atctcgtcca
     8581 gataatgctt gtgctgctcc acaaacagct gtttctgttc gttatcctct ggagatcctt
     8641 tcagcttctc gtagtggctt gccagataca ggaagttcac gtacttagaa ggcagggcca
     8701 gttcattgcc tttctgcagc tctccagcgc ttgccagcat gcgtttcctg ccgttttcca
     8761 gctcaaacag actgtacttg ggcagtttaa tgatcaggtc tttcttcact tccttataac
     8821 ctttggcctc cagaaaatcg attgggttct tctcgaaaga gcttctctcc ataattgtga
     8881 tacccagcag ctccttgaca gatttcagtt tcttgctttt gcccttctcc accttagcga
     8941 ccaccaggac ggagtatgcc acagtgggag agtcaaagcc tccatacttc ttagggtccc
     9001 agtctttctt tctagcgatc agcttatcag agttgcgttt tggcagaata ctttcttttg
     9061 agaaaccgcc tgtctgcacc tcagttttct tgacgatatt cacttgaggc atgctcagga
     9121 ccttcctcac ggttgcaaaa tctctgcctt tgtcccacac gatttcgcct gtctctccgt
     9181 ttgtctcaat caggggcctt ttgcggattt ctccattggc cagggtgatc tctgtcttaa
     9241 agaaattcat gatgtttgag taaaagaaat acttggcagt agcttttcca atttcctgct
     9301 cggacttggc gatcattttt ctgacatcgt acaccttata gtctccgtac acgaattcgc
     9361 tctccagttt tgggtatttc ttgatcagtg cggtaccgac cacggcgttc aggtatgcgt
     9421 catgggcatg gtgataattg ttaatctcgc ggaccttata aaactggaaa tctttcctaa
     9481 agtcgctgac cagtttactc ttcagggtaa tgaccttcac ttctctgatc agtttgtcat
     9541 tctcatcgta ctttgtgttc atgcgactat ccagaatctg tgcgacgtgt ttagtgatct
     9601 ggcgtgtttc caccagctgc ctcttgatga atccagcttt gtccagttct gacagtccac
     9661 cgcgctctgc ttttgtcagg ttatcaaact tcctctgagt gatcagtttg gcattcagca
     9721 gctgtctcca gtagttcttc attttcttga ccacttcctc actagggaca ttatctgatt
     9781 ttcctctgtt tttgtcggac ctggtcagca ccttgttgtc aatgctgtca tctttgatga
     9841 aactctgggg gacgatatga tccacgtcgt aatcagacag cctgttaata tccagctcct
     9901 ggtccacgta catatctctt ccattctgca gatagtacag atacagcttc tcattctgca
     9961 gctgtgtgtt ttccactggg tgctctttca ggatctggct acccagctcc ttaatgcctt
    10021 cctcgattct tttcatcctc tctctactgt tcttctgacc cttctgggtt gtctgatttt
    10081 cgcgagccat ctcaatcacg atgttttcgg gcttatggcc catgactttc accagctcat
    10141 cgacgatttt cactgtctgc agaatgcctt tcttgatggc tgggcttcca gccaggtttg
    10201 caatctgctc atgcagggag tggccctgtc cagacacctg agccttctgg atgtcctctt
    10261 tgaaggtcag actgtcatcg tgaatcagct gcataaaatt gcggttggcg aagccatcac
    10321 ttttcagaaa gtccaggatt gtctttcctg actgtttatc cctaatgccg ttgatcagct
    10381 tcctgctcag tctaccccag ccagtgtagc gccttctctt cagctgtttc atcaccttgt
    10441 catcgaacag atgagcatag gttttcagtc tttcctcgat cattcctctg tcctcaaaca
    10501 gggtcagtgt cagcacaatg tcttccagga tatcctcatt ttcctcgttg tccaggaagt
    10561 ctttatcctt aatgatcttc agcagatcgt ggtaagcgcc caggcttgcg ttaaatctgt
    10621 cttcgactcc gctgatctcc acggaatcga agcattcaat tttcttgaag tagtcctcct
    10681 tcagctgctt gacagtcact ttcctgttgg ttttaaacag cagatccacg attgctttct
    10741 tctgctctcc tgacaggaag gctggctttc tcatgccctc ggtgacgtat ttcacctttg
    10801 tcagttcgtt atagacagtg aaatactcgt acagcaggga gtgcttaggc agcactttct
    10861 cattgggcag gtttttgtca aaattagtca ttctctcgat gaaggactgt gcagaggcgc
    10921 ccttatcgac cacttcctcg aagttccaag gagtaatggt ttcctcggat ttgcgagtca
    10981 tccaggcaaa tctgctgttt ccgcgagcca gtggacccac atagtaaggg attctgaagg
    11041 tcagaatttt ttcgatcttc tcgcggttat ctttcagaaa ggggtagaag tcttcctggc
    11101 gcctcaggat agcgtgcagc tctcccagat gaatctggtg tgggatggaa ccgttatcaa
    11161 atgtgcgctg cttcctcagc aggtcctctc tattcagttt caccagcagt tcctcagttc
    11221 cgtccatctt ctccagaata ggtttgataa acttgtagaa ttcctcctgt gatgcgcctc
    11281 cgtcgatata accggcgtag ccgttcttgg actgatcaaa gaaaatttcc ttgtacttct
    11341 ctggcagctg ctgcctgacc agagccttca gcagggtcag atcctgatgg tgctcgtcat
    11401 atctcttgat catgctggca ctcagaggtg cttttgtaat ctcgctgttc actctcagaa
    11461 tatctgacag caggattgcg tcggacagat ttttggcagc caggaacagg tcagcgtact
    11521 gatctccgat ctgtgccagc aggttatcca ggtcatcgtc atatgtgtcc tttgacagct
    11581 gcagtttggc gtcctcagcc agatcaaaat tgcttttgaa gttaggagtc aggcccagac
    11641 tcagagcgat cagatttcca aacagaccgt tcctcttttc tccgggcagc tgtgcaatca
    11701 ggttctccag tctgcgtgac ttggacagcc tggctgacag gatagctttt gcgtccactc
    11761 tggaagcatt aatagggttt tcctcaaaca gctgattgta aatctggacc agctggatga
    11821 acagcttgtc cacatcggaa ttgtcggggt tcagatcgcc ctcgatcaga aaatgacccc
    11881 tgaacttaat catgtgggcc agagccagat agatcagtct caggtcggct ttatcagtag
    11941 agtcagccag tttcttgcgc agatgataga tggttgggta tttttcgtga taggcgacct
    12001 cgtccacaat gtttccaaag ataggatggc gctcgtgttt cttatcttcc tcgaccagga
    12061 aactttcctc cagcctatga aagaatgaat cgtccacctt agccatttcg ttagagaaaa
    12121 tctcctgcag gtaacagatt ctattcttcc ttctggtata gcgccttctt gctgttcttt
    12181 tcaggcgtgt tgcttcggca gtctctccag aaccgaacag cagggctcca atcagatttt
    12241 tcttgatgct gtgcctgtcg gtgttaccca gcaccttaaa tttcttgcta gggaccttgt
    12301 agtcgtctgt aatcactgcc caacccacgg agttagttcc aatatccagg ccgatgctat
    12361 acttcttatc catgcttccc acctttctct tcttcttagg tggagaagcc atggtggcgt
    12421 cgacctcgag ggggggcccg gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
    12481 cactggccgt cgttttacaa gcctgctttt ttgtacaaac ttgcgttcaa ctatgtgtta
    12541 gcttcaaaaa agtattccct ttgtgaattt aggtgatgtg caagaactga ccagtcgtct
    12601 tggccagctg aagatgtcaa tgccaaagaa aagacacaaa atgtatttaa gggggtggga
    12661 ggggattgct cttctcatat tgtttttttt acccaaacga tgaatttatg cctttataaa
    12721 gttcatttga cgtcaccggt ccgattcgac aaaagcctct ttgagacgat gggttactag
    12781 tgtctaaact gaaactggct tacggctaac taacgtttca tctttagttc cactcatcct
    12841 gcataaaaca ttgtttgttc tcagcatccg gcatacaatc actagcactg ttggtctaaa
    12901 aacaagacag tttaagatgg agttatctct gaaatgttga agagcaaaag ataggaaaga
    12961 gaactttgtt gtctgatggg cctttggtaa ttgcattttt ttcttaagag ttgaaatatt
    13021 gtgcacacgt atgtaagtgc gcacaaacaa aatcacagac gtacagttca tattcagatt
    13081 catattcatt tttttttcta caatgatgcc tcatgtacat catcttatta tcttacgtgt
    13141 catttacagt caaaaaaaaa ctagctggtt gaataaaagt aactttaagt cagaatttgc
    13201 cagacgtatg aacactttgg ggcttgattg taagtctttt tttttttttt gttgttgttg
    13261 gatgtactgg cacaattatc atcagagtaa atcagtaact gttgtgcatt tgcacagaaa
    13321 gcatagaaat gaactgtagt taaaaaagct aacaatcata aacaaagtag catgtgcata
    13381 tactgtactg aatttttttt aaataataat tgacataatc actggttgta atgtgtaaaa
    13441 aatgagtctg ccttaaccca acttccactc taaattgtgt gttgatgctc atttcccaaa
    13501 agggctgaat tacgatagaa aagataagtc aacagtttgt tgtctctgca ttgtttgagt
    13561 tgactacatc ttgagcaaat gtagctattg taaaatgggg atgagtatgt ggtacagaga
    13621 accatctcgt caatggagtg gaaactgcat taacatagag agatagagag aacagctgtg
    13681 tctaggcatg gggtataaga ttctgatggt atgataacct ttgataaaaa tatcacagtt
    13741 tgatggtatt gtgattgcca ctctaaaata tattattttt tttaatgtct tagtaaaaaa
    13801 caaaaacttt aaacataata tattttattt tgagaaacat tgaaaaaata ttttggaaca
    13861 gaaaacatgt caagcttaat aatttcaatg aatgatcgac ttctgctgtc ttcattagtt
    13921 tcaaaaacag atttctttac aatttaaaaa gcatttttgt atctcttttc tgctgaaaat
    13981 gctgttgtcc taaaaaacaa aaaaataaaa aataaataaa aaatcttaaa tataccttag
    14041 gaacggtata gcagaaaata ttggtggttt taaacccttg actttttcaa accggggtat
    14101 accttgaaaa cggttatcgt cctatgccta gctttgtcaa caaccaaaaa gcaactttat
    14161 taacatctta aggcaagttt aaaggcaaca tctaaaacag aatgaacggt tggagtttag
    14221 tgaagtttat ttataaacta atttcgagag aatcacgtgc ttatgactga ctacagctgg
    14281 tcccacatta gcctatgtat gattcaccaa tcagacaatt cctaactcat cataggccgc
    14341 cagagttcct ttcatttcct cgaaagagcg gcacagcagc ccagttgtta gcactcacaa
    14401 cttttctata caaagttgat agcttggcgt aatcatggtc atagctgttt cctgctcgag
    14461 ggcccatctg gcctgtgttt cagacaccag ggagtctctg ctcacgtttc ctgctatttg
    14521 cagcctctct atcaagacta atacacctct tcccgcatcg gctgcctgtg agaggctttt
    14581 cagcactgca ggattgcttt tcagccccaa aagagctagg cttgacacta acaattttga
    14641 gaatcagctt ctactgaagt taaatctgag gttttacaac tttgagtagc gtgtactggc
    14701 attagattgt ctgtcttata gtttgataat taaatacaaa cagttctaaa gcaggataaa
    14761 accttgtatg catttcattt aatgtttttt gagattaaaa gcttaaacaa gaatctctag
    14821 ttttctttct tgcttttact tttacttcct taatactcaa gtacaatttt aatggagtac
    14881 ttttttactt ttactcaagt aagattctag ccagatactt ttacttttaa ttgagtaaaa
    14941 ttttccctaa gtacttgtac tttcacttga gtaaaatttt tgagtacttt ttacacctct
    15001 gtcaagaact cctggacaaa cctctgacct gtgtggaaca gagtggatat gggtgtctga
    15061 acagatattc acgtcttttg cagatcagag ggcatttctg gtgttgtgca atgacctggg
    15121 tccaacaggt aggcgtactc gtatccaaga gcattgttgg acaggaggta gtctccattc
    15181 ctataaagag gaaggaatgg caccatgtag tagccgtcat tgtggccaat tggtgcattg
    15241 gtacgtgggt agatggaccg gggaggctga tgagttctta gccagcgtgg gccatcgccc
    15301 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
    15361 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
    15421 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
    15481 tttaacaaaa tattaacgct tacaatttag gtgg
//
