LOCUS       sequence-238383-       16069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway cloning destination vector for expressing enhancer driven 
            Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16069)
  AUTHORS   Poe AR, Wang B, Sapar ML, Ji H, Li K, Onabajo T, Fazliyeva R, Gibbs 
            M, Qiu Y, Hu Y, Han C
  TITLE     Robust CRISPR/Cas9-Mediated Tissue Specific Mutagenesis Reveals Gene
            Redundancy and Perdurance in Drosophila.
  JOURNAL   Genetics. 2018 Nov 30. pii: genetics.118.301736. doi: 
            10.1534/genetics.118.301736.
  PUBMED    30504366
REFERENCE   2  (bases 1 to 16069)
  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: "Genetics. 
            2018 Nov 30. pii: genetics.118.301736. doi: 
            10.1534/genetics.118.301736."
COMMENT     Sequence Label: pDEST-APIC-Cas9
FEATURES             Location/Qualifiers
     source          1..16069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(1..31,32..233)
                     /label=P element 3' end
                     /note="P element 3' end"
     protein_bind    631..1016
                     /label=gypsy insulator
                     /bound_moiety="Su(Hw), CP190, andMod(mdg4)67.2"
                     /note="chromatin insulator from Drosophila"
     protein_bind    1051..1175
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1200..1205,1206..1223,1224..1230)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1284..1943
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1350..1369)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2285..2590
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2469..2488
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2631..2755)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3063..3083
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3108..7211
                     /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             3108..7211
                     /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             3108..7211
                     /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             7212..7259
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    7923..8308
                     /label=gypsy insulator
                     /bound_moiety="Su(Hw), CP190, andMod(mdg4)67.2"
                     /note="chromatin insulator from Drosophila"
     primer_bind     complement(8318..8336)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(8788..8859,9261..9534,9609..10263,10325..10640,
                     10844..10975,11046..11660)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This is a modified version of the white gene lacking
                     part of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     misc_feature    join(12439..12993,12994..13024)
                     /label=P element 5' end
     rep_origin      complement(13259..13847)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13339..13358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14018..14809,14810..14878))
                     /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     14641..14660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14879..14983)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     15051..15069
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(15107..15129)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     15229..15248
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    15457..15526
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
ORIGIN
        1 catgatgaaa taacataagg tggtcccgtc ggcaagagac atccacttaa cgtatgcttg
       61 caataagtgc gagtgaaagg aatagtattc tgagtgtcgt attgagtctg agtgagacag
      121 cgatatgatt gttgattaac ccttagcatg tccgtggggt ttgaattaac tcataatatt
      181 aattagacga aattattttt aaagttttat ttttaataat ttgcgagtac gcaaagcttc
      241 ctaggtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
      301 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
      361 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
      421 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
      481 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
      541 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
      601 agccgaactg ttcgccaagc ttggctgcat cacgtaataa gtgtgcgttg aatttattcg
      661 caaaaacatt gcatattttc ggcaaagtaa aattttgttg cataccttat caaaaaataa
      721 gtgctgcata ctttttagag aaaccaaata attttttatt gcatacccgt ttttaataaa
      781 atacattgca taccctcttt taataaaaaa tattgcatac tttgacgaaa caaattttcg
      841 ttgcataccc aataaaagat tattatattg catacccgtt tttaataaaa tacattgcat
      901 accctctttt aataaaaaat attgcatacg ttgacgaaac aaattttcgt tgcataccca
      961 ataaaagatt attatattgc ataccttttc ttgccatacc atttagccga tcaattgtgc
     1021 tcggcaacag catgctgcag aagacttatc acaagtttgt acaaaaaagc tgaacgagaa
     1081 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
     1141 taatactgta aaacacaaca tatccagtca ctatggcggc cgcattaggc accccaggct
     1201 ttacacttta tgcttccggc tcgtataatg tgtggatttt gagttaggat ccgtcgagat
     1261 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     1321 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     1381 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     1441 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     1501 tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     1561 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     1621 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     1681 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
     1741 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     1801 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     1861 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     1921 atgagtggca gggcggggcg taaacgcgtg gatccggctt actaaaagcc agataacagt
     1981 atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg tatacccgaa
     2041 gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca gtgacagttg acagcgacag
     2101 ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag cacaaccatg
     2161 cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg
     2221 gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa caggggctgg
     2281 tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga
     2341 tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc
     2401 acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga
     2461 aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga
     2521 agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg
     2581 gggaatataa atgtcaggct cccttataca cagccagtct gcaggtcgac catagtgact
     2641 ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata
     2701 tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa gtggtgattt
     2761 gtcaaggtac cgagcgccgg agtataaata gaggcgcttc gtctacggag ctcagttgtg
     2821 aatgaatgtt cgagccgagc agacgtgccg cgcgaaagct aagcaaataa acaagcgcag
     2881 ctgagcaagc taaacaatcg taaagtgcaa gttaaagtga atcaattaaa agtaaccagc
     2941 aaccaagtaa atcaactgca actactgaaa tctgccggtg agattgtaca cccttcttaa
     3001 agatcccctt caaagtctaa attcttcctt ttcgtttagc cactcgagaa atcaaaatgg
     3061 ccccaaagaa gaagcggaag gtcggtatcc acggtgtccc agcagccatg gacaagaagt
     3121 actccattgg gctcgatatc ggcacaaaca gcgtcggctg ggccgtcatt acggacgagt
     3181 acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac agcataaaga
     3241 agaacctcat tggcgccctc ctgttcgact ccggggagac ggccgaagcc acgcggctca
     3301 aaagaacagc acggcgcaga tatacccgca gaaagaatcg gatctgctac ctgcaggaga
     3361 tctttagtaa tgagatggct aaggtggatg actctttctt ccataggctg gaggagtcct
     3421 ttttggtgga ggaggataaa aagcacgagc gccacccaat ctttggcaat atcgtggacg
     3481 aggtggcgta ccatgaaaag tacccaacca tatatcatct gaggaagaag cttgtagaca
     3541 gtactgataa ggctgacttg cggttgatct atctcgcgct ggcgcatatg atcaaatttc
     3601 ggggacactt cctcatcgag ggggacctga acccagacaa cagcgatgtc gacaaactct
     3661 ttatccaact ggttcagact tacaatcagc ttttcgaaga gaacccgatc aacgcatccg
     3721 gagttgacgc caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg ctcgaaaacc
     3781 tcatcgcaca gctccctggg gagaagaaga acggcctgtt tggtaatctt atcgccctgt
     3841 cactcgggct gacccccaac tttaaatcta acttcgacct ggccgaagat gccaagcttc
     3901 aactgagcaa agacacctac gatgatgatc tcgacaatct gctggcccag atcggcgacc
     3961 agtacgcaga cctttttttg gcggcaaaga acctgtcaga cgccattctg ctgagtgata
     4021 ttctgcgagt gaacacggag atcaccaaag ctccgctgag cgctagtatg atcaagcgct
     4081 atgatgagca ccaccaagac ttgactttgc tgaaggccct tgtcagacag caactgcctg
     4141 agaagtacaa ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga tacattgacg
     4201 gcggagcaag ccaggaggaa ttttacaaat ttattaagcc catcttggaa aaaatggacg
     4261 gcaccgagga gctgctggta aagcttaaca gagaagatct gttgcgcaaa cagcgcactt
     4321 tcgacaatgg aagcatcccc caccagattc acctgggcga actgcacgct atactcaggc
     4381 ggcaagagga tttctacccc tttttgaaag ataacaggga aaagattgag aaaatcctca
     4441 catttcggat accctactat gtaggccccc tcgcccgggg aaattccaga ttcgcgtgga
     4501 tgactcgcaa atcagaagag accatcactc cctggaactt cgaggaagtc gtggataagg
     4561 gggcctctgc ccagtccttc atcgaaagga tgactaactt tgataaaaat ctgcctaacg
     4621 aaaaggtgct tcctaaacac tctctgctgt acgagtactt cacagtttat aacgagctca
     4681 ccaaggtcaa atacgtcaca gaagggatga gaaagccagc attcctgtct ggagagcaga
     4741 agaaagctat cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg aaacagctca
     4801 aagaagacta tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc ggagtggagg
     4861 atcgcttcaa cgcatccctg ggaacgtatc acgatctcct gaaaatcatt aaagacaagg
     4921 acttcctgga caatgaggag aacgaggaca ttcttgagga cattgtcctc acccttacgt
     4981 tgtttgaaga tagggagatg attgaagaac gcttgaaaac ttacgctcat ctcttcgacg
     5041 acaaagtcat gaaacagctc aagaggcgcc gatatacagg atgggggcgg ctgtcaagaa
     5101 aactgatcaa tgggatccga gacaagcaga gtggaaagac aatcctggat tttcttaagt
     5161 ccgatggatt tgccaaccgg aacttcatgc agttgatcca tgatgactct ctcaccttta
     5221 aggaggacat ccagaaagca caagtttctg gccaggggga cagtcttcac gagcacatcg
     5281 ctaatcttgc aggtagccca gctatcaaaa agggaatact gcagaccgtt aaggtcgtgg
     5341 atgaactcgt caaagtaatg ggaaggcata agcccgagaa tatcgttatc gagatggccc
     5401 gagagaacca aactacccag aagggacaga agaacagtag ggaaaggatg aagaggattg
     5461 aagagggtat aaaagaactg gggtcccaaa tccttaagga acacccagtt gaaaacaccc
     5521 agcttcagaa tgagaagctc tacctgtact acctgcagaa cggcagggac atgtacgtgg
     5581 atcaggaact ggacatcaat cggctctccg actacgacgt ggatcatatc gtgccccagt
     5641 cttttctcaa agatgattct attgataata aagtgttgac aagatccgat aaaaatagag
     5701 ggaagagtga taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat tattggcggc
     5761 agctgctgaa cgccaaactg atcacacaac ggaagttcga taatctgact aaggctgaac
     5821 gaggtggcct gtctgagttg gataaagcag gcttcatcaa aaggcagctt gttgagacac
     5881 gccagatcac caagcacgtg gcccaaattc tcgattcacg catgaacacc aagtacgatg
     5941 aaaatgacaa actgattcga gaggtgaaag ttattactct gaagtctaag ctggtctcag
     6001 atttcagaaa ggactttcag ttttataagg tgagagagat caacaattac caccatgcgc
     6061 atgatgccta cctgaatgca gtggtaggca ctgcacttat caaaaaatat cccaagcttg
     6121 aatctgaatt tgtttacgga gactataaag tgtacgatgt taggaaaatg atcgcaaagt
     6181 ctgagcagga aataggcaag gccaccgcta agtacttctt ttacagcaat attatgaatt
     6241 ttttcaagac cgagattaca ctggccaatg gagagattcg gaagcgacca cttatcgaaa
     6301 caaacggaga aacaggagaa atcgtgtggg acaagggtag ggatttcgcg acagtccgga
     6361 aggtcctgtc catgccgcag gtgaacatcg ttaaaaagac cgaagtacag accggaggct
     6421 tctccaagga aagtatcctc ccgaaaagga acagcgacaa gctgatcgca cgcaaaaaag
     6481 attgggaccc caagaaatac ggcggattcg attctcctac agtcgcttac agtgtactgg
     6541 ttgtggccaa agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag gaactgctgg
     6601 gcatcacaat catggagcga tcaagcttcg aaaaaaaccc catcgacttt ctcgaggcga
     6661 aaggatataa agaggtcaaa aaagacctca tcattaagct tcccaagtac tctctctttg
     6721 agcttgaaaa cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag aaaggtaacg
     6781 agctggcact gccctctaaa tacgttaatt tcttgtatct ggccagccac tatgaaaagc
     6841 tcaaagggtc tcccgaagat aatgagcaga agcagctgtt cgtggaacaa cacaaacact
     6901 accttgatga gatcatcgag caaataagcg aattctccaa aagagtgatc ctcgccgacg
     6961 ctaacctcga taaggtgctt tctgcttaca ataagcacag ggataagccc atcagggagc
     7021 aggcagaaaa cattatccac ttgtttactc tgaccaactt gggcgcgcct gcagccttca
     7081 agtacttcga caccaccata gacagaaagc ggtacacctc tacaaaggag gtcctggacg
     7141 ccacactgat tcatcagtca attacggggc tctatgaaac aagaatcgac ctctctcagc
     7201 tcggtggaga caagcgtcct gctgctacta agaaagctgg tcaagctaag aaaaagaaat
     7261 aattaattaa ctaagccagt cggcaatcgg acgccttcga aacatgcaac actaatgttt
     7321 aattcagatt tcagcagaga caagctaaaa caccgacgag ttgtaatcat ttctgtgcgc
     7381 cagcatatat ttcttatata caacgtaata cataattatg taattctagc atctccccaa
     7441 cactcacata caaacaaaca aaaaatacaa acacacaaaa cgtatttacc cgcacgcatc
     7501 cttggcgagg ttgagtatga aacaaaaaca aaacttaatt tagagcaaag taattacacg
     7561 aataaattta ataaaaaaaa actataataa aaacgccaat catgttattt ccaaaaaaaa
     7621 aaaaaaaaaa aaanctagac gagatgtttg cgcttgtgtc gatgaatgta aagaattgca
     7681 actacataga tggtaattaa tttcagctgt aagaagtaca agtttttcat aattgtgaga
     7741 aaagtgcagt agtatcgctt aaagttcagt tttaatcagg gcatttagca gctcgaaccc
     7801 aatgtatttg taagcccaat aaacgtgtaa atgcctagag caaaatatca caactgtagc
     7861 ttttcatccg aaagataata atgaatagac tgacctgacg tttttcaaaa tgcgtaacta
     7921 gtcacgtaat aagtgtgcgt tgaatttatt cgcaaaaaca ttgcatattt tcggcaaagt
     7981 aaaattttgt tgcatacctt atcaaaaaat aagtgctgca tactttttag agaaaccaaa
     8041 taatttttta ttgcataccc gtttttaata aaatacattg cataccctct tttaataaaa
     8101 aatattgcat actttgacga aacaaatttt cgttgcatac ccaataaaag attattatat
     8161 tgcatacccg tttttaataa aatacattgc ataccctctt ttaataaaaa atattgcata
     8221 cgttgacgaa acaaattttc gttgcatacc caataaaaga ttattatatt gcataccttt
     8281 tcttgccata ccatttagcc gatcaattct agtatgtatg taagttaata aaacccattt
     8341 ttgcggaaag tagataaaaa aaacattttt tttttttact gcactggata tcattgaact
     8401 tatctgatca gttttaaatt tacttcgatc caagggtatt tgatgtacca ggttctttcg
     8461 attacctctc actcaaaatg acattccact caaagtcagc gctgtttgcc tccttctctg
     8521 tccacagaaa tatcgccgtc tctttcgccg ctgcgtccgc tatctctttc gccaccgttt
     8581 gtagcgttac gtagcgtcaa tgtccgcctt cagttgcatt ttgtcagcgg tttcgtgacg
     8641 aagctccaag cggtttacgc catcaattaa acacaaagtg ctgtgccaaa actcctctcg
     8701 cttcttattt ttgtttgttt tttgagtgat tggggtggtg attggttttg ggtgggtaag
     8761 caggggaaag tgtgaaaaat cccggcaatg ggccaagagg atcaggagct attaattcgc
     8821 ggaggcagca aacacccatc tgccgagcat ctgaacaatg tgagtagtac atgtgcatac
     8881 atcttaagtt cacttgatct ataggaactg cgattgcaac atcaaattgt ctgcggcgtg
     8941 agaactgcga cccacaaaaa tcccaaaccg caattgcaca aacaaatagt gacacgaaac
     9001 agattattct ggtagctgtt ctcgctatat aagacaattt ttgagatcat atcatgatca
     9061 agacatctaa aggcattcat tttcgactat attctttttt acaaaaaata taacaaccag
     9121 atattttaag ctgatcctag atgcacaaaa aataaataaa agtataaacc tacttcgtag
     9181 gatacttcgg ggtacttttt gttcggggtt agatgagcat aacgcttgta gttgatattt
     9241 gagatcccct atcattgcag ggtgacagcg gagcggcttc gcagagctgc attaaccagg
     9301 gcttcgggca ggccaaaaac tacggcacgc tccggccacc cagtccgccg gaggactccg
     9361 gttcagggag cggccaacta gccgagaacc tcacctatgc ctggcacaat atggacatct
     9421 ttggggcggt caatcagccg ggctccggat ggcggcagct ggtcaaccgg acacgcggac
     9481 tattctgcaa cgagcgacac ataccggcgc ccaggaaaca tttgctcaag aacggtgagt
     9541 ttctattcgc agtcggctga tctgtgtgaa atcttaataa agggtccaat taccaatttg
     9601 aaactcagtt tgcggcgtgg cctatccggg cgaacttttg gccgtgatgg gcagttccgg
     9661 tgccggaaag acgaccctgc tgaatgccct tgcctttcga tcgccgcagg gcatccaagt
     9721 atcgccatcc gggatgcgac tgctcaatgg ccaacctgtg gacgccaagg agatgcaggc
     9781 caggtgcgcc tatgtccagc aggatgacct ctttatcggc tccctaacgg ccagggaaca
     9841 cctgattttc caagccatgg tgcggatgcc acgacatctg acctatcggc agcgagtggc
     9901 ccgcgtggat caggtgatcc aggagctttc gctcagcaaa tgtcagcaca cgatcatcgg
     9961 tgtgcccggc agggtgaaag gtctgtccgg cggagaaagg aagcgtctgg cattcgcctc
    10021 cgaggctcta accgatccgc cgcttctgat ctgcgatgag cccacctccg gactggactc
    10081 ctttaccgcc cacagcgtcg tccaggtgct gaagaagctg tcgcagaagg gcaagaccgt
    10141 catcctgacc attcatcagc cgtcttccga gctgtttgag ctctttgaca agatccttct
    10201 gatggccgag ggcagggtag ctttcttggg cactcccagc gaagccgtcg acttcttttc
    10261 ctagtgagtt cgatgtgttt attaagggta tctagtatta cataacatct caactcctat
    10321 ccagcgtggg tgcccagtgt cctaccaact acaatccggc ggacttttac gtacaggtgt
    10381 tggccgttgt gcccggacgg gagatcgagt cccgtgatcg gatcgccaag atatgcgaca
    10441 attttgccat tagcaaagta gcccgggata tggagcagtt gttggccacc aaaaatctgg
    10501 agaagccact ggagcagccg gagaatgggt acacctacaa ggccacctgg ttcatgcagt
    10561 tccgggcggt cctgtggcga tcctggctgt cggtgctcaa ggaaccactc ctcgtaaaag
    10621 tgcgacttat tcagacaacg gtgagtggtt ccagtggaaa caaatgatat aacgcttaca
    10681 attcttggaa acaaattcgc tagattttag atagaattgc ctgattccac acccttctta
    10741 gtttttttca atgagatgta tagtttatag ttttgcagaa gataaataaa tttcatttaa
    10801 ctcgcgaata ttaatgagat gcgagtaaca ttttaatttg cagatggttg ccatcttgat
    10861 tggcctcatc tttttgggcc aacaactcac gcaagtgggt gtgatgaata tcaacggagc
    10921 catcttcctc ttcctgacca acatgacctt tcaaaacgtc tttgccacga taaatgtaag
    10981 tcatgtttag aatacatttg catttcaata atttactaac tttctaatga atcgattcga
    11041 tttaggtgtt cacctcagag ctgccagttt ttatgaggga ggcccgaagt cgactttatc
    11101 gctgtgacac atactttctg ggcaaaacga ttgccgaatt gccgcttttt ctcacagtgc
    11161 cactggtctt cacggcgatt gcctatccga tgatcggact gcgggccgga gtgctgcact
    11221 tcttcaactg cctggcgctg gtcactctgg tggccaatgt gtcaacgtcc ttcggatatc
    11281 taatatcctg cgccagctcc tcgacctcga tggcgctgtc tgtgggtccg ccggttatca
    11341 taccattcct gctctttggc ggcttcttct tgaactcggg ctcggtgcca gtatacctca
    11401 aatggttgtc gtacctctca tggttccgtt acgccaacga gggtctgctg attaaccaat
    11461 gggcggacgt ggagccgggc gaaattagct gcacatcgtc gaacaccacg tgccccagtt
    11521 cgggcaaggt catcctggag acgcttaact tctccgccgc cgatctgccg ctggactacg
    11581 tgggtctggc cattctcatc gtgagcttcc gggtgctcgc atatctggct ctaagacttc
    11641 gggcccgacg caaggagtag ccgacatata tccgaaataa ctgcttgttt ttttttttta
    11701 ccattattac catcgtgttt actgtttatt gccccctcaa aaagctaatg taattatatt
    11761 tgtgccaata aaaacaagat atgacctata gaatacaagt atttcccctt cgaacatccc
    11821 cacaagtaga ctttggattt gtcttctaac caaaagactt acacacctgc ataccttaca
    11881 tcaaaaactc gtttatcgct acataaaaca ccgggatata ttttttatat acatactttt
    11941 caaatcgcgc gccctcttca taattcacct ccaccacacc acgtttcgta gttgctcttt
    12001 cgctgtctcc cacccgctct ccgcaacaca ttcacctttt gttcgacgac cttggagcga
    12061 ctgtcgttag ttccgcgcga ttcggttcgc tcaaatggtt ccgagtggtt catttcgtct
    12121 caatagaaat tagtaataaa tatttgtatg tacaatttat ttgctccaat atatttgtat
    12181 atatttccct cacagctata tttattctaa tttaatatta tgacttttta aggtaatttt
    12241 ttgtgacctg ttcggagtga ttagcgttac aatttgaact gaaagtgaca tccagtgttt
    12301 gttccttgtg tagatgcatc tcaaaaaaat ggtgggcata atagtgttgt ttatatatat
    12361 caaaaataac aactataata ataagaatac atttaattta gaaaatgctt ggatttcact
    12421 ggaactagaa ttaattcggc tgctgctcta aacgacgcat ttcgtactcc aaagtacgaa
    12481 ttttttccct caagctctta ttttcattaa acaatgaaca ggacctaacg cacagtcacg
    12541 ttattgttta cataaatgat tttttttact attcaaactt actctgtttg tgtactccca
    12601 ctggtatagc cttcttttat cttttctggt tcaggctcta tcactttact aggtacggca
    12661 tctgcgttga gtcgcctcct tttaaatgtc tgaccttttg caggtgcagc cttccactgc
    12721 gaatcattaa agtgggtatc acaaatttgg gagttttcac caaggctgca cccaaggctc
    12781 tgctcccaca attttctctt aatagcacac ttcggcacgt gaattaattt tactccagtc
    12841 acagctttgc agcaaaattt gcaatatttc attttttttt attccacgta agggttaatg
    12901 ttttcaaaaa aaaattcgtc cgcacacaac ctttcctctc aacaagcaaa cgtgcactga
    12961 atttaagtgt atacttcggt aagcttcggc tatcgacggg accaccttat gttatttcat
    13021 catgggccag acccacgtag tccagcggca gatcggcggc ggagaagtta agcgtctcca
    13081 ggatgacctt gcccgaactg gggcacgtgg tgttcgacga tgtgcagcta atttcgcccg
    13141 gctccacgtc cgcccattgg ttaatcagca gaccctcgtt ggcgtaacgg aaccatgaga
    13201 ggtacgacaa ccatttgagg tatactggca ccgagcccga gttcaagaag aaggcgtttt
    13261 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
    13321 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
    13381 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
    13441 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
    13501 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
    13561 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
    13621 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
    13681 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
    13741 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
    13801 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
    13861 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
    13921 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
    13981 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
    14041 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
    14101 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
    14161 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
    14221 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
    14281 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
    14341 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
    14401 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
    14461 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
    14521 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
    14581 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
    14641 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
    14701 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
    14761 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
    14821 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
    14881 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
    14941 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
    15001 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
    15061 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
    15121 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
    15181 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
    15241 agattgtact gagagtgcac cattcgacat gcccgccgtg accgtcgaga acccgctgac
    15301 gctgccccgc gtatccgcac ccgccgacgc cgtcgcacgt cccgtgctca ccgtgaccac
    15361 cgcgcccagc ggtttcgagg gcgagggctt cccggtgcgc cgcgcgttcg ccgggatcaa
    15421 ctaccgccac ctcgacccgt tcatcatgat ggaccagatg ggtgaggtgg agtacgcgcc
    15481 cggggagccc aagggcacgc cctggcaccc gcaccgcggc ttcgagaccg tgacctacat
    15541 cgtcgatatg cggtgtgaaa taccgcaccg aatcgcgcgg aactaacgac agtcgctcca
    15601 aggtcgtcga acaaaaggtg aatgtgttgc ggagagcggg tgggagacag cgaaagagca
    15661 actacgaaac gtggtgtggt ggaggtgaat tatgaagagg gcgcgcgatt tgaaaagtat
    15721 gtatataaaa aatatatccc ggtgttttat gtagcgataa acgagttttt gatgtaaggt
    15781 atgcaggtgt gtaagtcttt tggttagaag acaaatccaa agtctacttg tggggatgtt
    15841 cgaaggggaa atacttgtat tctataggtc atatcttgtt tttattggca caaatataat
    15901 tacattagct ttttgagggg gcaataaaca gtaaacacga tggtaataat ggtaaaaaaa
    15961 aaaaacaagc agttatttcg gatatatgtc ggctactcct tgcgtcgggc ccgaagtctt
    16021 agagccagat atgcgagcac ccggaagctc acgatgagaa tggccagac
//
