LOCUS       sequence-101869-       13981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Fok1:dCas9 under control of nanos promoter and 3'UTR. For 
            germ line restricted high specificity genome engineering in 
            Drosophila melanogaster..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13981)
  TITLE     CRISPRflydesign
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13981)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pnos-Fok1:dCas9-nos
FEATURES             Location/Qualifiers
     source          1..13981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            666..4802
                     /gene="white"
                     /label=mini-white
                     /note="This modified version of the white gene lacks part 
                     of the first intron."
     primer_bind     complement(675..693)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(1145..1216,1618..1891,1966..2620,2682..2997,3218..3349,
                     3420..4034)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This modified version of the white gene lacks part 
                     of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     protein_bind    4809..4842
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             5791..5811
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5818..6405
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     CDS             join(6421..6444,6445..6447,6448..8931,8932..8937,8938..9003,
                     9004..9006,9007..10521)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(6421..6444,6445..6447,6448..8934,8935..8937,
                     8938..10521)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6421..10521
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             10534..10554
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    11505..11574
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(11793..11811)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11832..11848)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11832..11848)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11845..11867)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11856..11872
                     /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)."
     promoter        complement(join(11880..11886,11887..11904,11905..11910))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11925..11946
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12063..12080)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12234..12822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12314..12333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12993..13784,13785..13853))
                     /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     13616..13635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13854..13958)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtacgtaccg
      661 ggcccctagt atgtatgtaa gttaataaaa cccatttttg cggaaagtag ataaaaaaaa
      721 catttttttt ttttactgca ctggatatca ttgaacttat ctgatcagtt ttaaatttac
      781 ttcgatccaa gggtatttga tgtaccaggt tctttcgatt acctctcact caaaatgaca
      841 ttccactcaa agtcagcgct gtttgcctcc ttctctgtcc acagaaatat cgccgtctct
      901 ttcgccgctg cgtccgctat ctctttcgcc accgtttgta gcgttacgta gcgtcaatgt
      961 ccgccttcag ttgcattttg tcagcggttt cgtgacgaag ctccaagcgg tttacgccat
     1021 caattaaaca caaagtgctg tgccaaaact cctctcgctt cttatttttg tttgtttttt
     1081 gagtgattgg ggtggtgatt ggttttgggt gggtaagcag gggaaagtgt gaaaaatccc
     1141 ggcaatgggc caagaggatc aggagctatt aattcgcgga ggcagcaaac acccatctgc
     1201 cgagcatctg aacaatgtga gtagtacatg tgcatacatc ttaagttcac ttgatctata
     1261 ggaactgcga ttgcaacatc aaattgtctg cggcgtgaga actgcgaccc acaaaaatcc
     1321 caaaccgcaa ttgcacaaac aaatagtgac acgaaacaga ttattctggt agctgttctc
     1381 gctatataag acaatttttg agatcatatc atgatcaaga catctaaagg cattcatttt
     1441 cgactatatt cttttttaca aaaaatataa caaccagata ttttaagctg atcctagatg
     1501 cacaaaaaat aaataaaagt ataaacctac ttcgtaggat acttcggggt actttttgtt
     1561 cggggttaga tgagcataac gcttgtagtt gatatttgag atcccctatc attgcagggt
     1621 gacagcggag cggcttcgca gagctgcatt aaccagggct tcgggcaggc caaaaactac
     1681 ggcacgctcc ggccacccag tccgccggag gactccggtt cagggagcgg ccaactagcc
     1741 gagaacctca cctatgcctg gcacaatatg gacatctttg gggcggtcaa tcagccgggc
     1801 tccggatggc ggcagctggt caaccggaca cgcggactat tctgcaacga gcgacacata
     1861 ccggcgccca ggaaacattt gctcaagaac ggtgagtttc tattcgcagt cggctgatct
     1921 gtgtgaaatc ttaataaagg gtccaattac caatttgaaa ctcagtttgc ggcgtggcct
     1981 atccgggcga acttttggcc gtgatgggca gttccggtgc cggaaagacg accctgctga
     2041 atgcccttgc ctttcgatcg ccgcagggca tccaagtatc gccatccggg atgcgactgc
     2101 tcaatggcca acctgtggac gccaaggaga tgcaggccag gtgcgcctat gtccagcagg
     2161 atgacctctt tatcggctcc ctaacggcca gggaacacct gattttccag gccatggtgc
     2221 ggatgccacg acatctgacc tatcggcagc gagtggcccg cgtggatcag gtgatccagg
     2281 agctttcgct cagcaaatgt cagcacacga tcatcggtgt gcccggcagg gtgaaaggtc
     2341 tgtccggcgg agaaaggaag cgtctggcat tcgcctccga ggcactaacc gatccgccgc
     2401 ttctgatctg cgatgagccc acctccggac tggactcatt taccgcccac agcgtcgtcc
     2461 aggtgctgaa gaagctgtcg cagaagggca agaccgtcat cctgaccatt catcagccgt
     2521 cttccgagct gtttgagctc tttgacaaga tccttctgat ggccgagggc agggtagctt
     2581 tcttgggcac tcccagcgaa gccgtcgact tcttttccta gtgagttcga tgtgtttatt
     2641 aagggtatct agcattacat tacatctcaa ctcctatcca gcgtgggtgc ccagtgtcct
     2701 accaactaca atccggcgga cttttacgta caggtgttgg ccgttgtgcc cggacgggag
     2761 atcgagtccc gtgatcggat cgccaagata tgcgacaatt ttgctattag caaagtagcc
     2821 cgggatatgg agcagttgtt ggccaccaaa aatttggaga agccactgga gcagccggag
     2881 aatgggtaca cctacaaggc cacctggttc atgcagttcc gggcggtcct gtggcgatcc
     2941 tggctgtcgg tgctcaagga accactcctc gtaaaagtgc gacttattca gacaacggtg
     3001 agtggttcca gtggaaacaa atgatataac gcttacaatt cttggaaaca aattcgctag
     3061 attttagtta gaattgcctg attccacacc cttcttagtt tttttcaatg agatgtatag
     3121 tttatagttt tgcagaaaat aaataaattt catttaactc gcgaacatgt tgaagatatg
     3181 aatattaatg agatgcgagt aacattttaa tttgcagatg gttgccatct tgattggcct
     3241 catctttttg ggccaacaac tcacgcaagt gggcgtgatg aatatcaacg gagccatctt
     3301 cctcttcctg accaacatga cctttcaaaa cgtctttgcc acgataaatg taagtcttgt
     3361 ttagaataca tttgcatatt aataatttac taactttcta atgaatcgat tcgatttagg
     3421 tgttcacctc agagctgcca gtttttatga gggaggcccg aagtcgactt tatcgctgtg
     3481 acacatactt tctgggcaaa acgattgccg aattaccgct ttttctcaca gtgccactgg
     3541 tcttcacggc gattgcctat ccgatgatcg gactgcgggc cggagtgctg cacttcttca
     3601 actgcctggc gctggtcact ctggtggcca atgtgtcaac gtccttcgga tatctaatat
     3661 cctgcgccag ctcctcgacc tcgatggcgc tgtctgtggg tccgccggtt atcataccat
     3721 tcctgctctt tggcggcttc ttcttgaact cgggctcggt gccagtatac ctcaaatggt
     3781 tgtcgtacct ctcatggttc cgttacgcca acgagggtct gctgattaac caatgggcgg
     3841 acgtggagcc gggcgaaatt agctgcacat cgtcgaacac cacgtgcccc agttcgggca
     3901 aggtcatcct ggagacgctt aacttctccg ccgccgatct gccgctggac tacgtgggtc
     3961 tggccattct catcgtgagc ttccgggtgc tcgcatatct ggctctaaga cttcgggccc
     4021 gacgcaagga gtagccgaca tatatccgaa ataactgctt gttttttttt tttaccatta
     4081 ttaccatcgt gtttactgtt tattgccccc tcaaaaagct aatgtaatta tatttgtgcc
     4141 aataaaaaca agatatgacc tatagaatac aagtatttcc ccttcgaaca tccccacaag
     4201 tagactttgg atttgtcttc taaccaaaag acttacacac ctgcatacct tacatcaaaa
     4261 actcgtttat cgctacataa aacaccggga tatatttttt atatacatac ttttcaaatc
     4321 gcgcgccctc ttcataattc acctccacca caccacgttt cgtagttgct ctttcgctgt
     4381 ctcccacccg ctctccgcaa cacattcacc ttttgttcga cgaccttgga gcgactgtcg
     4441 ttagttccgc gcgattcggt tcgctcaaat ggttccgagt ggttcatttc gtctcaatag
     4501 aaattagtaa taaatatttg tatgtacaat ttatttgctc caatatattt gtatatattt
     4561 ccctcacagc tatatttatt ctaatttaat attatgactt tttaaggtaa ttttttgtga
     4621 cctgttcgga gtgattagcg ttacaatttg aactgaaagt gacatccagt gtttgttcct
     4681 tgtgtagatg catctcaaaa aaatggtggg cataatagtg ttgtttatat atatcaaaaa
     4741 taacaactat aataataaga atacatttaa tttagaaaat gcttggattt cactggaact
     4801 aggctagcat aacttcgtat aatgtatgct atacgaagtt atgctagcgg atccaagctt
     4861 cgaccgtttt aacctcgaaa tatgcacatg taaggacgga tgtgagcgaa cgccagtgat
     4921 gaccgggatc agaggtaacc taccatgcat ggtggggatt aggtgaccgt tcgcaggtag
     4981 tttgatcgga gcgaatgttc ggggggtctg gcgtcagagg ctctaaactt tatgtaattc
     5041 ctgccgcgaa acacgcacgt atcaagcagt cagctgttct cttcgttcag cgcgcgccgg
     5101 tgttgcaaaa cgagcgctct tcgccggcgg tggctcgtgc gatagttcgt tttgtcggta
     5161 atccgatgtt gccgcgccga tatcatgtga tgttgtcaca gtgcgcgaaa ttcgaatggt
     5221 ggtgtgcagt gattgtgttg tgacggcgag tggcgcgtgt gggtgcttag ttttgggaga
     5281 tgttttcgta tttttttgtt gataactcag gctttgttgc tgtgttgtag tactattttc
     5341 cattgcgcgg tgtccagctt ttaattagtg gcacatattc ttagcaagta aaaattattt
     5401 tgcatactat taaatttctt ataaattatt ttctaaaatt aagtttacct tttcaatttt
     5461 actaaaaata tcgatatatt tattatcgct ggaaaactac attattccac ctctaagcaa
     5521 gaaccgttag ttggcgcgta gctttaccac aaaattcctg gaattgccgt acgcttcgca
     5581 gttgtttcaa gttgtctaag ggacatacga ttttttttgc ctctgcgtca cgattttaac
     5641 ccaaaagcga gtttagttac atgtacatta ttattagata aagaagtatc gcgaatactt
     5701 cagttgaata aactgtgctt ggtttttggg tgaggatttg tggaaagtag agtgcgcgat
     5761 aaccgtaact ttcgacccgg attttcgatg cctaagaaga agcggaaggt gagcagccaa
     5821 cttgtgaagt ctgaactcga ggagaaaaaa tcagagttga gacacaagtt gaagtacgtg
     5881 ccacacgaat acatcgagct tatcgagatc gccagaaaca gtacccagga taggatcctt
     5941 gagatgaaag tcatggagtt ctttatgaag gtctacggtt atagaggaaa gcaccttggc
     6001 ggtagcagaa agcccgatgg cgccatctat actgtcggat ctcctatcga ttatggggtg
     6061 atcgtggata ccaaagctta ctcaggcggg tacaacttgc ccataggaca agccgacgag
     6121 atgcagcggt atgtcgaaga gaaccagacg cgcaacaagc acatcaaccc caatgaatgg
     6181 tggaaagtgt acccaagtag tgtgactgag ttcaagttcc tgtttgtctc cggccacttt
     6241 aagggcaatt ataaagctca gctcactaga ctcaatcaca tcacaaactg caacggagct
     6301 gtgttgtcag tggaggagct cctgattgga ggcgagatga tcaaagccgg cacccttaca
     6361 ctggaggagg tgcggcggaa gttcaacaat ggagagatca acttcggtgg cggtggatcc
     6421 gacaagaagt actccattgg gctcgctatc ggcacaaaca gcgtcggctg ggccgtcatt
     6481 acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac
     6541 agcataaaga agaacctcat tggcgccctc ctgttcgact ccggggagac ggccgaagcc
     6601 acgcggctca aaagaacagc acggcgcaga tatacccgca gaaagaatcg gatctgctac
     6661 ctgcaggaga tctttagtaa tgagatggct aaggtggatg actctttctt ccataggctg
     6721 gaggagtcct ttttggtgga ggaggataaa aagcacgagc gccacccaat ctttggcaat
     6781 atcgtggacg aggtggcgta ccatgaaaag tacccaacca tatatcatct gaggaagaag
     6841 cttgtagaca gtactgataa ggctgacttg cggttgatct atctcgcgct ggcgcatatg
     6901 atcaaatttc ggggacactt cctcatcgag ggggacctga acccagacaa cagcgatgtc
     6961 gacaaactct ttatccaact ggttcagact tacaatcagc ttttcgaaga gaacccgatc
     7021 aacgcatccg gagttgacgc caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg
     7081 ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga acggcctgtt tggtaatctt
     7141 atcgccctgt cactcgggct gacccccaac tttaaatcta acttcgacct ggccgaagat
     7201 gccaagcttc aactgagcaa agacacctac gatgatgatc tcgacaatct gctggcccag
     7261 atcggcgacc agtacgcaga cctttttttg gcggcaaaga acctgtcaga cgccattctg
     7321 ctgagtgata ttctgcgagt gaacacggag atcaccaaag ctccgctgag cgctagtatg
     7381 atcaagcgct atgatgagca ccaccaagac ttgactttgc tgaaggccct tgtcagacag
     7441 caactgcctg agaagtacaa ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga
     7501 tacattgacg gcggagcaag ccaggaggaa ttttacaaat ttattaagcc catcttggaa
     7561 aaaatggacg gcaccgagga gctgctggta aagcttaaca gagaagatct gttgcgcaaa
     7621 cagcgcactt tcgacaatgg aagcatcccc caccagattc acctgggcga actgcacgct
     7681 atcctcaggc ggcaagagga tttctacccc tttttgaaag ataacaggga aaagattgag
     7741 aaaatcctca catttcggat accctactat gtaggccccc tcgcccgggg aaattccaga
     7801 ttcgcgtgga tgactcgcaa atcagaagag accatcactc cctggaactt cgaggaagtc
     7861 gtggataagg gggcctctgc ccagtccttc atcgaaagga tgactaactt tgataaaaat
     7921 ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt acgagtactt cacagtttat
     7981 aacgagctca ccaaggtcaa atacgtcaca gaagggatga gaaagccagc attcctgtct
     8041 ggagagcaga agaaagctat cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg
     8101 aaacagctca aagaagacta tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc
     8161 ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc acgatctcct gaaaatcatt
     8221 aaagacaagg acttcctgga caatgaggag aacgaggaca ttcttgagga cattgtcctc
     8281 acccttacgt tgtttgaaga tagggagatg attgaagaac gcttgaaaac ttacgctcat
     8341 ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc gatatacagg atgggggcgg
     8401 ctgtcaagaa aactgatcaa tgggatccga gacaagcaga gtggaaagac aatcctggat
     8461 tttcttaagt ccgatggatt tgccaaccgg aacttcatgc agttgatcca tgatgactct
     8521 ctcaccttta aggaggacat ccagaaagca caagtttctg gccaggggga cagtcttcac
     8581 gagcacatcg ctaatcttgc aggtagccca gctatcaaaa agggaatact gcagaccgtt
     8641 aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata agcccgagaa tatcgttatc
     8701 gagatggccc gagagaacca aactacccag aagggacaga agaacagtag ggaaaggatg
     8761 aagaggattg aagagggtat aaaagaactg gggtcccaaa tccttaagga acacccagtt
     8821 gaaaacaccc agcttcagaa tgagaagctc tacctgtact acctgcagaa cggcagggac
     8881 atgtacgtgg atcaggaact ggacatcaat cggctctccg actacgacgt ggatgccatc
     8941 gtgccccagt cttttctcaa agatgattct attgataata aagtgttgac aagatccgat
     9001 aaaaatagag ggaagagtga taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat
     9061 tattggcggc agctgctgaa cgccaaactg atcacacaac ggaagttcga taatctgact
     9121 aaggctgaac gaggtggcct gtctgagttg gataaagccg gcttcatcaa aaggcagctt
     9181 gttgagacac gccagatcac caagcacgtg gcccaaattc tcgattcacg catgaacacc
     9241 aagtacgatg aaaatgacaa actgattcga gaggtgaaag ttattactct gaagtctaag
     9301 ctggtctcag atttcagaaa ggactttcag ttttataagg tgagagagat caacaattac
     9361 caccatgcgc atgatgccta cctgaatgca gtggtaggca ctgcacttat caaaaaatat
     9421 cccaagcttg aatctgaatt tgtttacgga gactataaag tgtacgatgt taggaaaatg
     9481 atcgcaaagt ctgagcagga aataggcaag gccaccgcta agtacttctt ttacagcaat
     9541 attatgaatt ttttcaagac cgagattaca ctggccaatg gagagattcg gaagcgacca
     9601 cttatcgaaa caaacggaga aacaggagaa atcgtgtggg acaagggtag ggatttcgcg
     9661 acagtccgga aggtcctgtc catgccgcag gtgaacatcg ttaaaaagac cgaagtacag
     9721 accggaggct tctccaagga aagtatcctc ccgaaaagga acagcgacaa gctgatcgca
     9781 cgcaaaaaag attgggaccc caagaaatac ggcggattcg attctcctac agtcgcttac
     9841 agtgtactgg ttgtggccaa agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag
     9901 gaactgctgg gcatcacaat catggagcga tcaagcttcg aaaaaaaccc catcgacttt
     9961 ctcgaggcga aaggatataa agaggtcaaa aaagacctca tcattaagct tcccaagtac
    10021 tctctctttg agcttgaaaa cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag
    10081 aaaggtaacg agctggcact gccctctaaa tacgttaatt tcttgtatct ggccagccac
    10141 tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga agcagctgtt cgtggaacaa
    10201 cacaaacact accttgatga gatcatcgag caaataagcg aattctccaa aagagtgatc
    10261 ctcgccgacg ctaacctcga taaggtgctt tctgcttaca ataagcacag ggataagccc
    10321 atcagggagc aggcagaaaa cattatccac ttgtttactc tgaccaactt gggcgcgcct
    10381 gcagccttca agtacttcga caccaccata gacagaaagc ggtacacctc tacaaaggag
    10441 gtcctggacg ccacactgat tcatcagtca attacggggc tctatgaaac aagaatcgac
    10501 ctctctcagc tcggtggaga cagcagggct gaccccaaga agaagaggaa ggtgtgagcg
    10561 aatccagctc tggagcagag gctctggcag cttttgcagc gtttatataa catgaaatat
    10621 atatacgcat tccgatcaaa gctgggttaa ccagatagat agtaacgttt aactagcgcc
    10681 tggcgcgttc gattttaaag agatttagag cgttatcccg tgcctataga tcttatagta
    10741 tagacaacga acgatcactc aaatccaagt caataattca agaatttatg tctgtttctg
    10801 tgaaagggaa actaattttg ttaaagaaga cttacaatat cgtaatactt gttcaatcgt
    10861 cgtggccgat agaaatatct tacaatccga aagttgatga atggaattgg tctgcaactg
    10921 gtcgccttca ttttgtaaaa tgttcgcttg cggccgaaaa atttcgatat atctacaatt
    10981 gatatacaat ctttactaaa ttttgaaaaa ggaacacttt gaatttcgaa ctgtcaatcg
    11041 tatcattaga atttaatcta aatttaaatc ttgctaaagg aaatagcaag gaacactttc
    11101 gtcgtcggct acgcattcat tgtaaaattt taaattttga cattccgcac tttttgatag
    11161 ataagcgaag agtattttta ttacatgtat cgcaagtatt catttcaaca cacatatcta
    11221 tatatatatg ttatatattt attcaatttt gtttaccatt gatcaatttt tcacacatga
    11281 aacaaccgcc agcattatat aattttttta tttttttaaa aaatgtgtac acatattctg
    11341 aaaatgaaaa attcaatggc tcgagtgcca aataaagaaa tggttacaat ttaaggaaac
    11401 aaatgtcctt cttgcgtttg aaacaactaa tccttttcgc cctcgcggcg tttctcgaaa
    11461 agggccagga aggatccact agtgtcgacg atgtaggtca cggtctcgaa gccgcggtgc
    11521 gggtgccagg gcgtgccctt gggctccccg ggcgcgtact ccacctcacc catctggtcc
    11581 atcatgatga acgggtcgag gtggcggtag ttgatcccgg cgaacgcgcg gcgcaccggg
    11641 aagccctcgc cctcgaaacc gctgggcgcg gtggtcacgg tgagcacggg acgtgcgacg
    11701 gcgtcggcgg gtgcggatac gcggggcagc gtcagcgggt tctcgacggt cacggcgggc
    11761 atgtcgacac tagttctagc cagcttttgt tccctttagt gagggttaat ttcgagcttg
    11821 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
    11881 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
    11941 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
    12001 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
    12061 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
    12121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
    12181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
    12241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
    12301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
    12361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
    12421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
    12481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
    12541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
    12601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
    12661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
    12721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
    12781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
    12841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
    12901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
    12961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
    13021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
    13081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
    13141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
    13201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
    13261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
    13321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
    13381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
    13441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
    13501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
    13561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
    13621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
    13681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
    13741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
    13801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
    13861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
    13921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
    13981 c
//
