LOCUS       sequence-331633-       14781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For germline transformation. Expresses Cas9 in germ cells and 
            ZsGreen marker in all cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14781)
  AUTHORS   Kandul NP, Belikoff EJ, Liu J, Buchman A, Li F, Yamamoto A, Yang T, 
            Shriner I, Scott MJ, Akbari OS
  TITLE     Genetically Encoded CRISPR Components Yield Efficient Gene Editing 
            in the Invasive Pest Drosophila suzukii.
  JOURNAL   CRISPR J. 2021 Oct;4(5):739-751. doi: 10.1089/crispr.2021.0032.
  PUBMED    34661429
REFERENCE   2  (bases 1 to 14781)
  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"; doi: 
            "10.1089/crispr.2021"; journalName: "CRISPR J"; date: "2021-10"; 
            volume: "4"; issue: "5"; pages: "739-751"
COMMENT     Sequence Label: pB[vasa-Cas9 PUb-ZsGreen]
FEATURES             Location/Qualifiers
     source          1..14781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(270..286)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     307..323
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2574..6677
                     /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             2574..6677
                     /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             2574..6677
                     /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             6690..6710
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8089..8784)
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein (Matz et al., 
                     1999)"
                     /label=ZsGreen
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIADYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     protein_bind    10923..11022
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     repeat_region   11286..11348
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(11478..11495)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11478..11494)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11487..11509)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      11636..12091
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11723..11742)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11933..11954
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        12117..12221
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12222..12290,12291..13082)
                     /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(12440..12459)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      13253..13841
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13742..13761
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13995..14012
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    14129..14150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(14165..14170,14171..14188,14189..14195)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14203..14219
                     /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     14208..14230
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     14227..14243
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14227..14243
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   14734..14768
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 ttgctctctc tttctaaata gcgcgaatcc gtcgctgtgc atttaggaca tctcagtcgc
       61 cgcttggagc tcccgtgagg cgtgcttgtc aatgcggtaa gtgtcactga ttttgaacta
      121 taacgaccgc gtgagtcaaa atgacgcatg attatctttt acgtgacttt taagatttaa
      181 ctcatacgat aattatattg ttatttcatg ttctacttac gtgataactt attatatata
      241 tattttcttg ttatagatat caagcttatc gataccgtcg acctcgaggg ggggcccccc
      301 ctcgatcgag gtcgacggta tcgataagct tgatatcgaa ttcctgcagc tggttgtagg
      361 tgcagttgcg cttcttgcac ttggcgcact tgaggagatc ggtcttggtg ccctgcacgg
      421 tggccagctg ggcatcgttg atggcctcct tgacgaactt ctcgcgcagc ttcttcatct
      481 cgtcgctggc catctcctcc ggcgtcattt tggccagctg cttggcagtg acggcgccgc
      541 acataaagtt gccgcgcaat ccaggattct tggggtcctt cagattggcc acgcgtgacc
      601 taatgcgatt cttgtacttc atatccgtgt tattgaactc ggagtaaatg gcatcctcca
      661 gttcggcggc catttcctcc ggctcaccgc atccctcggg cacttcgcca atcttcagtg
      721 cggtggccag catttcgcgg catttgatgc ggaccgcatc tgtcatgcca ccggagggaa
      781 acgaggtctg tgacgaggat gtcgagcctt tcttctcttt atccttggat gagctggatg
      841 aactggactt atccttgccg gagattgacg acgacgactt ggcggcactg gtggacttcg
      901 aggcactgct gttgttggag gagccctcct tagcggagct gttgttcggt gtggttggcg
      961 ccgggctggc caggaagcgt ttccagttct tgatcagcgt cttggccaga gcgatcacct
     1021 cgtcgtcctt gctactcttg cgcagctcgt ttacggtcat gccgatgcgc gttttggtca
     1081 gaatgtcgag attgatgtta agcgtttgca gggccttcag caggtccaga gcctgatcct
     1141 gtccctgtac ggtggatata tgtatatttg tgacatgatg cgcggcacgt tagctaatcc
     1201 cgatacttac tgtgccgtcg ctggccatct tgctcatctt cttttggatt cgaaacactt
     1261 cctcctccac gctcattttg gcccacttaa tttgatcaga ggttaggtta actggcgtcg
     1321 gctgtcggct ttaatcggtg ctgcgggtct gcgacagttg tttatgcgga agccgggcta
     1381 tttctttttt tccctccaat aaatgttgtt tactattttt ttcgtttctg tattcgcaga
     1441 acaagttgac agaacaccct aacgcgtttt agtgttaccg ggcggtggct tgtcaagaaa
     1501 taccttccca tggctaatac aattcccaac tggaagtagt ttgggtttca gaagtcaaca
     1561 gtgttaccgt attttaatgt acatctggct tttaagcgag tacacacaca tagcattgcg
     1621 atgtttttta aacttgcact gcttgcaagc aggggtccac tgtatttcaa tgaaagaaca
     1681 gcgtgttggg cgtttttgaa ttatactaat taccttcaat aacaattccc tatatatcac
     1741 ttagttttaa taaataaatc gtttgagtgt ggtctagaga tgatttatta tgatgaagtg
     1801 cggttgttgg ggacaagcgg gatacgtctg cgggtcaaat cggcggtggg acatagtgag
     1861 agggcaacgg cgaacaatca ccaacgcgcc ctggtacgcg agaaatgttg atatcgcaca
     1921 tgacctcagc atccaccttc agtttgagaa caaattaaat caggcacata ccagacaacc
     1981 aggagaacac tgaggaggcg tcatcaaaga tatctttgga cacgtggcat aaacaagcca
     2041 aactattatc taatcaatat ttttatacaa attttgtatg tttccatgtt atcttcgttg
     2101 ctcctgttag ttactacgtt tcatagccct taaattggtt gcttaacgta aaataaaatt
     2161 atattgtata aaaataaaga ggaattcacc tataacaaac gaaaaagtaa ataaataaga
     2221 acttccccat gttaaaaatg ccaccaccat ctttacatat aagtatgtac atatgaatga
     2281 atcacttagg ttgcttgaat attacaattt cattaatagc taaatcacat ttgatgtgtt
     2341 agtggaaaac ggctatatat ataaattatc gaaattgtga atatcgaatt gcgatagcac
     2401 aatgggaaat tccaccacta gatttttggt acttttaaca gatccttttc ggttttgcgt
     2461 tgcgcgaagt gatctgaact tatcaaagtt tgtaaggtaa tacataaagt gaaaaagaat
     2521 taatttgctc ttgaaaggca ggccaaatta aaaaaaaaat atcaatggta cccatggaca
     2581 agaagtactc cattgggctc gatatcggca caaacagcgt cggctgggcc gtcattacgg
     2641 acgagtacaa ggtgccgagc aaaaaattca aagttctggg caataccgat cgccacagca
     2701 taaagaagaa cctcattggc gccctcctgt tcgactccgg ggagacggcc gaagccacgc
     2761 ggctcaaaag aacagcacgg cgcagatata cccgcagaaa gaatcggatc tgctacctgc
     2821 aggagatctt tagtaatgag atggctaagg tggatgactc tttcttccat aggctggagg
     2881 agtccttttt ggtggaggag gataaaaagc acgagcgcca cccaatcttt ggcaatatcg
     2941 tggacgaggt ggcgtaccat gaaaagtacc caaccatata tcatctgagg aagaagcttg
     3001 tagacagtac tgataaggct gacttgcggt tgatctatct cgcgctggcg catatgatca
     3061 aatttcgggg acacttcctc atcgaggggg acctgaaccc agacaacagc gatgtcgaca
     3121 aactctttat ccaactggtt cagacttaca atcagctttt cgaagagaac ccgatcaacg
     3181 catccggagt tgacgccaaa gcaatcctga gcgctaggct gtccaaatcc cggcggctcg
     3241 aaaacctcat cgcacagctc cctggggaga agaagaacgg cctgtttggt aatcttatcg
     3301 ccctgtcact cgggctgacc cccaacttta aatctaactt cgacctggcc gaagatgcca
     3361 agcttcaact gagcaaagac acctacgatg atgatctcga caatctgctg gcccagatcg
     3421 gcgaccagta cgcagacctt tttttggcgg caaagaacct gtcagacgcc attctgctga
     3481 gtgatattct gcgagtgaac acggagatca ccaaagctcc gctgagcgct agtatgatca
     3541 agcgctatga tgagcaccac caagacttga ctttgctgaa ggcccttgtc agacagcaac
     3601 tgcctgagaa gtacaaggaa attttcttcg atcagtctaa aaatggctac gccggataca
     3661 ttgacggcgg agcaagccag gaggaatttt acaaatttat taagcccatc ttggaaaaaa
     3721 tggacggcac cgaggagctg ctggtaaagc ttaacagaga agatctgttg cgcaaacagc
     3781 gcactttcga caatggaagc atcccccacc agattcacct gggcgaactg cacgctatcc
     3841 tcaggcggca agaggatttc tacccctttt tgaaagataa cagggaaaag attgagaaaa
     3901 tcctcacatt tcggataccc tactatgtag gccccctcgc ccggggaaat tccagattcg
     3961 cgtggatgac tcgcaaatca gaagagacca tcactccctg gaacttcgag gaagtcgtgg
     4021 ataagggggc ctctgcccag tccttcatcg aaaggatgac taactttgat aaaaatctgc
     4081 ctaacgaaaa ggtgcttcct aaacactctc tgctgtacga gtacttcaca gtttataacg
     4141 agctcaccaa ggtcaaatac gtcacagaag ggatgagaaa gccagcattc ctgtctggag
     4201 agcagaagaa agctatcgtg gacctcctct tcaagacgaa ccggaaagtt accgtgaaac
     4261 agctcaaaga agactatttc aaaaagattg aatgtttcga ctctgttgaa atcagcggag
     4321 tggaggatcg cttcaacgca tccctgggaa cgtatcacga tctcctgaaa atcattaaag
     4381 acaaggactt cctggacaat gaggagaacg aggacattct tgaggacatt gtcctcaccc
     4441 ttacgttgtt tgaagatagg gagatgattg aagaacgctt gaaaacttac gctcatctct
     4501 tcgacgacaa agtcatgaaa cagctcaaga ggcgccgata tacaggatgg gggcggctgt
     4561 caagaaaact gatcaatggg atccgagaca agcagagtgg aaagacaatc ctggattttc
     4621 ttaagtccga tggatttgcc aaccggaact tcatgcagtt gatccatgat gactctctca
     4681 cctttaagga ggacatccag aaagcacaag tttctggcca gggggacagt cttcacgagc
     4741 acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg aatactgcag accgttaagg
     4801 tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc cgagaatatc gttatcgaga
     4861 tggcccgaga gaaccaaact acccagaagg gacagaagaa cagtagggaa aggatgaaga
     4921 ggattgaaga gggtataaaa gaactggggt cccaaatcct taaggaacac ccagttgaaa
     4981 acacccagct tcagaatgag aagctctacc tgtactacct gcagaacggc agggacatgt
     5041 acgtggatca ggaactggac atcaatcggc tctccgacta cgacgtggat catatcgtgc
     5101 cccagtcttt tctcaaagat gattctattg ataataaagt gttgacaaga tccgataaaa
     5161 atagagggaa gagtgataac gtcccctcag aagaagttgt caagaaaatg aaaaattatt
     5221 ggcggcagct gctgaacgcc aaactgatca cacaacggaa gttcgataat ctgactaagg
     5281 ctgaacgagg tggcctgtct gagttggata aagccggctt catcaaaagg cagcttgttg
     5341 agacacgcca gatcaccaag cacgtggccc aaattctcga ttcacgcatg aacaccaagt
     5401 acgatgaaaa tgacaaactg attcgagagg tgaaagttat tactctgaag tctaagctgg
     5461 tctcagattt cagaaaggac tttcagtttt ataaggtgag agagatcaac aattaccacc
     5521 atgcgcatga tgcctacctg aatgcagtgg taggcactgc acttatcaaa aaatatccca
     5581 agcttgaatc tgaatttgtt tacggagact ataaagtgta cgatgttagg aaaatgatcg
     5641 caaagtctga gcaggaaata ggcaaggcca ccgctaagta cttcttttac agcaatatta
     5701 tgaatttttt caagaccgag attacactgg ccaatggaga gattcggaag cgaccactta
     5761 tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa gggtagggat ttcgcgacag
     5821 tccggaaggt cctgtccatg ccgcaggtga acatcgttaa aaagaccgaa gtacagaccg
     5881 gaggcttctc caaggaaagt atcctcccga aaaggaacag cgacaagctg atcgcacgca
     5941 aaaaagattg ggaccccaag aaatacggcg gattcgattc tcctacagtc gcttacagtg
     6001 tactggttgt ggccaaagtg gagaaaggga agtctaaaaa actcaaaagc gtcaaggaac
     6061 tgctgggcat cacaatcatg gagcgatcaa gcttcgaaaa aaaccccatc gactttctcg
     6121 aggcgaaagg atataaagag gtcaaaaaag acctcatcat taagcttccc aagtactctc
     6181 tctttgagct tgaaaacggc cggaaacgaa tgctcgctag tgcgggcgag ctgcagaaag
     6241 gtaacgagct ggcactgccc tctaaatacg ttaatttctt gtatctggcc agccactatg
     6301 aaaagctcaa agggtctccc gaagataatg agcagaagca gctgttcgtg gaacaacaca
     6361 aacactacct tgatgagatc atcgagcaaa taagcgaatt ctccaaaaga gtgatcctcg
     6421 ccgacgctaa cctcgataag gtgctttctg cttacaataa gcacagggat aagcccatca
     6481 gggagcaggc agaaaacatt atccacttgt ttactctgac caacttgggc gcgcctgcag
     6541 ccttcaagta cttcgacacc accatagaca gaaagcggta cacctctaca aaggaggtcc
     6601 tggacgccac actgattcat cagtcaatta cggggctcta tgaaacaaga atcgacctct
     6661 ctcagctcgg tggagacagc agggctgacc ccaagaagaa gaggaaggtg tgataagttt
     6721 aaacgcgcgc ttcgatcacg tgggcaaagc ccccgaccga tgacgatgaa gatgacgccc
     6781 aggatggcac cgaagacgta gcggcctagc tcgagaatgt atggacatag atttcaaata
     6841 attaaatgta atgcagtaat tgatgtaatt agttaaataa gttagatatt aataacatat
     6901 taattatatg tattataacg catataataa taaaatgcat atttaggata tgcaagccat
     6961 tcgaattttc tattttaatt tccttttaca aagaaatgta taacaaaata taatttgaaa
     7021 aaatgttctg gctctaattc gatttctttt aagtattttg tgaactgctt ttaataacga
     7081 gcggttgcaa aacttaacag aaacgtgcac tttgatccca actaataccg tgtactatca
     7141 cgtgtttggt tttaagtact catccttcgt gtttcgttgt ttgttgtttg ccttgctgta
     7201 cacttggctc ttgcgctctc tcgctctccg ttggagccgg ctttttgaat gcatgcctcg
     7261 ccctgctcgc gcccagtcct cctcccccga aaagcatgcc gaccagcaaa tgttgccctt
     7321 ttgcgtttgc tgtctttgca gaagcaaaat caataactga gaaatccacc acactgctgc
     7381 tcttcgtgtt ggatccacta gaactcgaat cgctatccaa gccagctcca ttgtcggcat
     7441 cgtgctcatt ctattgacgc tggcagattt ggttttggcg ctatgggacc cgttcggtta
     7501 caacaacatg tttccgcgcg agtttcccga cgacatgtcg cgcacgttcc tgactgcgta
     7561 ctttgagagt ttcgacaaca ccacgtccag agaaatcata gagtttatgc ccgagttctt
     7621 ttcggaaatg gtcgaaacgg acgatgacgc cacgtttgaa tctctatttc atttattaga
     7681 ttatgtggca tctttagaag ttaattccga cggccaaatg ttaaacttgg aggagggtga
     7741 tgaaattgag gattttgacg aatctacttt ggtggggcaa gcgttagcca ctagctcgct
     7801 atacactcgc atggagttta tgcagtacac gtttaggcaa aacacactat tgtctatgaa
     7861 caaagaaaac aacaatttta atcaaataat catgggttta tttgcaacaa acacaattgt
     7921 ggcgtttaca gcatttgtta tacacacaga actcatattt tttatatttt tcgtaatctt
     7981 cctaatgatc acattttatt acataatcaa agaatcgtac gaatattata aaacaattga
     8041 tttgttattt taaaaacgat tcattctaga gggtttggct gcaagctttc agggcaatgc
     8101 agatccggat gcaatagcat gttctgtcag atgccatttc tgattcttag catcgctgcg
     8161 gtcttcacgg gtgagcttat gctggatgaa gtgccagtcc ggcatctttc ttggcacaga
     8221 ctttgctttg taaactgtgt cgaattggca ccgtaaacgc ccaccatcct tcagaaggag
     8281 gtacatggag acatcccctt tcaatatccc ctgcttaggt actggtatga tcttctcgca
     8341 ggatggctcc cagttatctg tcatcttttt catcacaggt ccatcagcag gaaaattcac
     8401 tccataaaat ttggactcat gatacatgca gttttcttca acactcactg ttatatctgc
     8461 attacatatg caaactgctc catcctcaaa gagaaaagac ctgtcccatg tataaccagc
     8521 aggacacgag ttcttgaaat agtcagctat gtcttgagga tattcagtga aaaccctgtt
     8581 tccgtacata aaggcagctg acaatatgtc ttcggcaaat ggcaatggtc caccttcgac
     8641 cacacacaga ttaatagcct gtttcccttt gaacggatat ccaatgccct ctcccgtgat
     8701 cacaaattta tgtccatcga cgcacccttc catacggtat ttcattgtca tttcttttgt
     8761 tagaccgtgc tttgactgag ccatggatcc ccgatctgca ttttggatta ttctgcgggc
     8821 agaaaataga gatgtggaaa attagtacga aatcaaatga gtttcgttga aattacaaaa
     8881 ctattgaaac taacttcctg gctggggaat aaaaatggga aacttattta tcgacgccaa
     8941 ctttgttgag aaacccctat taaccctcta cgaatattgg aacaaaggaa agcgaagaaa
     9001 cagggaacaa agggtagttg agaaaccttt gttccgttgc ttcgtcatcg ttttcgtgtt
     9061 gcgagtgtgt gcatgtatat atacacagct gaaatgcatg catacacatt attttgtgtg
     9121 tatatggtga cgtcacaact acaagcaata agaaattttc agacgtggct ttgtttaagc
     9181 aacctactct atttcagcta aaaataagtg gatttcgttg gtaaaatact tcaattaact
     9241 aaacaactaa ctaataacat gcacacaaat gctcgagtgc gttcgtgatt tctcgaattt
     9301 tcaaatgcgt cactgcgaat ttcacaattt gccaataaat cttggcgaaa atcaacacgc
     9361 aagttttatt tatagatttg tttgcgtttt gatgccaatt cattgggaaa acaagatgcg
     9421 tggctgccaa tttcttattt tgtaattacg tagagcgttg aataaaaaaa aaatggccga
     9481 acaaagacct tgaaatgcag tttttcttga aattactcaa cgtcttgttg ctcttattac
     9541 taattggtaa cagcgagtta aaaacttacg tttcttgtga ctttcgagaa tgttctttta
     9601 attgtacttt aatcaccaac aattaagtat aaatttttcg ctgattgcgc tttactttct
     9661 acttgtactt gctgctgcaa atgtcaattg gttttgaagg cgaccgttcg cgaacgctgt
     9721 ttatatacct tcggtgtccg ttgaaaatca ctaaaaaata ccgtagtgtt cgtaacactt
     9781 tagtacagag aaaaaaaatt gtgccgaaat gtttttgata cgtacgaata ccttgtatta
     9841 aaatttttta tgatttctgt gtatcacttt ttttttgtgt ttttcgttta aactcaccac
     9901 agtacaaaac aataaaatat ttttaagaca atttcaaatt gagacctttc tcgtactgac
     9961 ttgaccggct gaatgaggat ttctacctag acgacctact tcttaccatg acattgaatg
    10021 caatgccacc tttgatctaa acttacaaaa gtccaaggct tgttaggatt ggtgttttgg
    10081 aatttaagac ccttgttagt ttgcttttga aatagcactg tcttctctac cggctataat
    10141 tttgaaactc gcagcttgac tggaaattta aaaagtaatt ctgtgtaggt aaagggtgtt
    10201 ttaaaagtgt gatgtgttga gcgttgcggc aacgactgct atttatgtat atattttcaa
    10261 aacttattgt ttttgaagtg ttttaaatgg agctatctgg caacgctgcg cataatctta
    10321 cacaagcttt tcttaatcca tttttaagtg aaatttgttt atactctttc ggcaaataat
    10381 tgttaaatcg ctttaagtgg gcttacatct ggataagtaa tgaaaacctg catattataa
    10441 tattaaaaca tataatccac tgtgctttcc ccgtgtgtgg ccatatacct aaaaaagttt
    10501 attttcgcag agccccgcac ggtcacacta cggttcggcg attttcgatt ttggacagta
    10561 ctgattgcaa gcgcaccgaa agcaaaatgg agctggagat tttgaacgcg aagaacagca
    10621 agccgtacgg caaggtgaag gtgccctccg gcgccacgcc catcggcgat ctgcgcgccc
    10681 taattcacaa gaccctgaag cagaccccac acgcgaatcg ccagtcgctt cgtctggaac
    10741 tgaagggcaa aagcctgaaa gatacggaca cattggaatc tctgtcgctg cgttccggcg
    10801 acaagatcgg gtaccgtcga ctgcagaatt cgaagcttga gctcgagatc ccccgggctg
    10861 tactagtcgc gctcgcgcga ctgacggtcg taagcacccg cgtacgtgtc caccccggtc
    10921 acaacccctt gtgtcatgtc ggcgacccta cgcccccaac tgagagaact caaaggttac
    10981 cccagttggg gcactactcc cgaaaaccgc ttctgacctg ggaaaacgtg aagccccggg
    11041 gcatccgctg agggttgccg ccggggcttc ggtgtgtccg tcagtactag tcccgggctg
    11101 caggaattcg ataaaagttt tgttacttta tagaagaaat tttgagtttt tgtttttttt
    11161 aataaataaa taaacataaa taaattgttt gttgaattta ttattagtat gtaagtgtaa
    11221 atataataaa acttaatatc tattcaaatt aataaataaa cctcgatata cagaccgata
    11281 aaacacatgc gtcaatttta cacatgatta tctttaacgt acgtcacaat atgattatct
    11341 ttctagggtt aatctagctg cgtgttctgc agcgtgtcga gcatcttcat ctgctccatc
    11401 acgctgtaaa acacatttgc accgcgagtc tgcccgtcct ccacgggttc aaaaacgtga
    11461 atgaacgagg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
    11521 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
    11581 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
    11641 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
    11701 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
    11761 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
    11821 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
    11881 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
    11941 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
    12001 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
    12061 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
    12121 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
    12181 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
    12241 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
    12301 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
    12361 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
    12421 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
    12481 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
    12541 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
    12601 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
    12661 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
    12721 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
    12781 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
    12841 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
    12901 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
    12961 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
    13021 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
    13081 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
    13141 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
    13201 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
    13261 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
    13321 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
    13381 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
    13441 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
    13501 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
    13561 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
    13621 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
    13681 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
    13741 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
    13801 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
    13861 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
    13921 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
    13981 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
    14041 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
    14101 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
    14161 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
    14221 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcccgccgg gtaactcacg
    14281 gggtatccat gtccatttct gcggcatcca gccaggatac ccgtcctcgc tgacgtaata
    14341 tcccagcgcc gcaccgctgt cattaatctg cacaccggca cggcagttcc ggctgtcgcc
    14401 ggtattgttc gggttgctga tgcgcttcgg gctgaccatc cggaactgtg tccggaaaag
    14461 ccgcgacgaa ctggtatccc aggtggcctg aacgaacagt tcaccgttaa aggcgtgcat
    14521 ggccacacct tcccgaatca tcatggtaaa cgtgcgtttt cgctcaacgt caatgcagca
    14581 gcagtcatcc tcggcaaact ctttccatgc cgcttcaacc tcgcgggaaa aggcacgggc
    14641 ttcttcctcc ccgatgccca gatagcgcca gcttgggcga tgactgagcc ggaaaaaaga
    14701 cccgacgata tgatcctgat gcagctagat taaccctaga aagatagtct gcgtaaaatt
    14761 gacgcatgca ttcttgaaat a
//
