LOCUS       sequence-331621-       14140 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 14140)
  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 14140)
  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[nos-Cas9 PUb-ZsGreen]
FEATURES             Location/Qualifiers
     source          1..14140
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(312..328)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     345..361
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   644..678
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(1169..1185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1169..1185)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1182..1204)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1193..1209
                     /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(1217..1223,1224..1241,1242..1247))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1262..1283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1400..1417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1571..2159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1651..1670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2330..3121,3122..3190))
                     /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     2953..2972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3191..3295)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3321..3776)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3458..3479)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3670..3689
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3903..3925
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3917..3934
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3918..3934
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   4064..4126
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    4390..4489
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     CDS             6628..7323
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein (Matz et al., 
                     1999)"
                     /label=ZsGreen
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIADYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     regulatory      9301..9310
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9307..13410
                     /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             9307..13410
                     /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             9307..13410
                     /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             13423..13443
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 tatatatata tatatatata tatatatata tgttatatat ttattcaatt ttgtttacca
       61 ttgatcaatt tttcacacat gaaacaaccg ccagcattat ataatttttt tattttttta
      121 aaaaatgtgt acacatattc tgaaaatgaa aaattcaatg gctcgagtgc caaataaaga
      181 aatggttaca atttaaggaa acaaatgtcc ttcttgcgtt tgaaacaact aatccttttc
      241 gccctcgcgg cgtttctcga aaagggccag gaagatgcca tcgcatgcac ttggctttcc
      301 accgttggta tcgataccgt cgacctcgag ggggggcccc ccctcgaggt cgacggtatc
      361 gataagcttg atatctataa caagaaaata tatatataat aagttatcac gtaagtagaa
      421 catgaaataa caatataatt atcgtatgag ttaaatctta aaagtcacgt aaaagataat
      481 catgcgtcat tttgactcac gcggtcgtta tagttcaaaa tcagtgacac ttaccgcatt
      541 gacaagcacg cctcacggga gctccaagcg gcgactgaga tgtcctaaat gcacagcgac
      601 ggattcgcgc tatttagaaa gagagagcaa tatttcaaga atgcatgcgt caattttacg
      661 cagactatct ttctagggtt aatctagctg catcaggatc atatcgtcgg gtcttttttc
      721 cggctcagtc atcgcccaag ctggcgctat ctgggcatcg gggaggaaga agcccgtgcc
      781 ttttcccgcg aggttgaagc ggcatggaaa gagtttgccg aggatgactg ctgctgcatt
      841 gacgttgagc gaaaacgcac gtttaccatg atgattcggg aaggtgtggc catgcacgcc
      901 tttaacggtg aactgttcgt tcaggccacc tgggatacca gttcgtcgcg gcttttccgg
      961 acacagttcc ggatggtcag cccgaagcgc atcagcaacc cgaacaatac cggcgacagc
     1021 cggaactgcc gtgccggtgt gcagattaat gacagcggtg cggcgctggg atattacgtc
     1081 agcgaggacg ggtatcctgg ctggatgccg cagaaatgga catggatacc ccgtgagtta
     1141 cccggcgggc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     1201 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     1261 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     1321 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     1381 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1441 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1501 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1561 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1621 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1681 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1741 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1801 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1861 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1921 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1981 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2041 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2101 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2161 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2221 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2281 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2341 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2401 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2461 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     2521 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     2581 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     2641 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     2701 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     2761 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     2821 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     2881 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     2941 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3001 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3061 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3121 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3181 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3241 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3301 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     3361 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     3421 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     3481 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     3541 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     3601 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3661 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3721 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     3781 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     3841 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     3901 ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt
     3961 tgaacccgtg gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca
     4021 gatgaagatg ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca
     4081 tattgtgacg tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct
     4141 gtatatcgag gtttatttat taatttgaat agatattaag ttttattata tttacactta
     4201 catactaata ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac
     4261 aaaaactcaa aatttcttct ataaagtaac aaaactttta tcgaattcct gcagcccggg
     4321 actagtactg acggacacac cgaagccccg gcggcaaccc tcagcggatg ccccggggct
     4381 tcacgttttc ccaggtcaga agcggttttc gggagtagtg ccccaactgg ggtaaccttt
     4441 gagttctctc agttgggggc gtagggtcgc cgacatgaca caaggggttg tgaccggggt
     4501 ggacacgtac gcgggtgctt acgaccgtca gtcgcgcgag cgcgactagt acagcccggg
     4561 ggatctcgag ctcaagcttc gaattctgca gtcgacggta cccgatcttg tcgccggaac
     4621 gcagcgacag agattccaat gtgtccgtat ctttcaggct tttgcccttc agttccagac
     4681 gaagcgactg gcgattcgcg tgtggggtct gcttcagggt cttgtgaatt agggcgcgca
     4741 gatcgccgat gggcgtggcg ccggagggca ccttcacctt gccgtacggc ttgctgttct
     4801 tcgcgttcaa aatctccagc tccattttgc tttcggtgcg cttgcaatca gtactgtcca
     4861 aaatcgaaaa tcgccgaacc gtagtgtgac cgtgcggggc tctgcgaaaa taaacttttt
     4921 taggtatatg gccacacacg gggaaagcac agtggattat atgttttaat attataatat
     4981 gcaggttttc attacttatc cagatgtaag cccacttaaa gcgatttaac aattatttgc
     5041 cgaaagagta taaacaaatt tcacttaaaa atggattaag aaaagcttgt gtaagattat
     5101 gcgcagcgtt gccagatagc tccatttaaa acacttcaaa aacaataagt tttgaaaata
     5161 tatacataaa tagcagtcgt tgccgcaacg ctcaacacat cacactttta aaacaccctt
     5221 tacctacaca gaattacttt ttaaatttcc agtcaagctg cgagtttcaa aattatagcc
     5281 ggtagagaag acagtgctat ttcaaaagca aactaacaag ggtcttaaat tccaaaacac
     5341 caatcctaac aagccttgga cttttgtaag tttagatcaa aggtggcatt gcattcaatg
     5401 tcatggtaag aagtaggtcg tctaggtaga aatcctcatt cagccggtca agtcagtacg
     5461 agaaaggtct caatttgaaa ttgtcttaaa aatattttat tgttttgtac tgtggtgagt
     5521 ttaaacgaaa aacacaaaaa aaaagtgata cacagaaatc ataaaaaatt ttaatacaag
     5581 gtattcgtac gtatcaaaaa catttcggca caattttttt tctctgtact aaagtgttac
     5641 gaacactacg gtatttttta gtgattttca acggacaccg aaggtatata aacagcgttc
     5701 gcgaacggtc gccttcaaaa ccaattgaca tttgcagcag caagtacaag tagaaagtaa
     5761 agcgcaatca gcgaaaaatt tatacttaat tgttggtgat taaagtacaa ttaaaagaac
     5821 attctcgaaa gtcacaagaa acgtaagttt ttaactcgct gttaccaatt agtaataaga
     5881 gcaacaagac gttgagtaat ttcaagaaaa actgcatttc aaggtctttg ttcggccatt
     5941 ttttttttat tcaacgctct acgtaattac aaaataagaa attggcagcc acgcatcttg
     6001 ttttcccaat gaattggcat caaaacgcaa acaaatctat aaataaaact tgcgtgttga
     6061 ttttcgccaa gatttattgg caaattgtga aattcgcagt gacgcatttg aaaattcgag
     6121 aaatcacgaa cgcactcgag catttgtgtg catgttatta gttagttgtt tagttaattg
     6181 aagtatttta ccaacgaaat ccacttattt ttagctgaaa tagagtaggt tgcttaaaca
     6241 aagccacgtc tgaaaatttc ttattgcttg tagttgtgac gtcaccatat acacacaaaa
     6301 taatgtgtat gcatgcattt cagctgtgta tatatacatg cacacactcg caacacgaaa
     6361 acgatgacga agcaacggaa caaaggtttc tcaactaccc tttgttccct gtttcttcgc
     6421 tttcctttgt tccaatattc gtagagggtt aataggggtt tctcaacaaa gttggcgtcg
     6481 ataaataagt ttcccatttt tattccccag ccaggaagtt agtttcaata gttttgtaat
     6541 ttcaacgaaa ctcatttgat ttcgtactaa ttttccacat ctctattttc tgcccgcaga
     6601 ataatccaaa atgcagatcg gggatccatg gctcagtcaa agcacggtct aacaaaagaa
     6661 atgacaatga aataccgtat ggaagggtgc gtcgatggac ataaatttgt gatcacggga
     6721 gagggcattg gatatccgtt caaagggaaa caggctatta atctgtgtgt ggtcgaaggt
     6781 ggaccattgc catttgccga agacatattg tcagctgcct ttatgtacgg aaacagggtt
     6841 ttcactgaat atcctcaaga catagctgac tatttcaaga actcgtgtcc tgctggttat
     6901 acatgggaca ggtcttttct ctttgaggat ggagcagttt gcatatgtaa tgcagatata
     6961 acagtgagtg ttgaagaaaa ctgcatgtat catgagtcca aattttatgg agtgaatttt
     7021 cctgctgatg gacctgtgat gaaaaagatg acagataact gggagccatc ctgcgagaag
     7081 atcataccag tacctaagca ggggatattg aaaggggatg tctccatgta cctccttctg
     7141 aaggatggtg ggcgtttacg gtgccaattc gacacagttt acaaagcaaa gtctgtgcca
     7201 agaaagatgc cggactggca cttcatccag cataagctca cccgtgaaga ccgcagcgat
     7261 gctaagaatc agaaatggca tctgacagaa catgctattg catccggatc tgcattgccc
     7321 tgaaagcttg cagccaaacc ctctagaatg aatcgttttt aaaataacaa atcaattgtt
     7381 ttataatatt cgtacgattc tttgattatg taataaaatg tgatcattag gaagattacg
     7441 aaaaatataa aaaatatgag ttctgtgtgt ataacaaatg ctgtaaacgc cacaattgtg
     7501 tttgttgcaa ataaacccat gattatttga ttaaaattgt tgttttcttt gttcatagac
     7561 aatagtgtgt tttgcctaaa cgtgtactgc ataaactcca tgcgagtgta tagcgagcta
     7621 gtggctaacg cttgccccac caaagtagat tcgtcaaaat cctcaatttc atcaccctcc
     7681 tccaagttta acatttggcc gtcggaatta acttctaaag atgccacata atctaataaa
     7741 tgaaatagag attcaaacgt ggcgtcatcg tccgtttcga ccatttccga aaagaactcg
     7801 ggcataaact ctatgatttc tctggacgtg gtgttgtcga aactctcaaa gtacgcagtc
     7861 aggaacgtgc gcgacatgtc gtcgggaaac tcgcgcggaa acatgttgtt gtaaccgaac
     7921 gggtcccata gcgccaaaac caaatctgcc agcgtcaata gaatgagcac gatgccgaca
     7981 atggagctgg cttggatagc gattcgagtt aaacgcgccg atttcagggc atccgattct
     8041 tccagcttct gggcctcctc acggaacttt cggatattgt ccttgatctc cttgttctta
     8101 tccatctccg ccttcatgtt gtcgaagaac tgcgagaaga agccggccct gcgacctggc
     8161 gcactataaa agcggggctg cggagtaaag tcatgactta ggccggttgc gccatggggt
     8221 tttatgagcc aaactctaac ctgctgctgc tgcaggttct gcgaggcctg ctggcacgtg
     8281 aagagacagg cgcgatcccg cacaagggca gctattctgt actgcaagga gaaaaaatca
     8341 ttgaaagctt cgaccgtttt aacctcgaaa tatgcacatg taaggacgga tgtgagcgaa
     8401 cgccagtgat gaccgggatc agaggtaacc taccatggtg gggattaggt gaccgttcgc
     8461 aggtagtttg atcggagcga atgttcgggg ggtctggcgt cagaggctct aaactttatg
     8521 taattcctgc cgcgaaacac gcacgtatca agcagtcagc tgttctcttc gttcagcgcg
     8581 cgccggtgtt gcaaaacgag cgctcttcgc cggcggtggc tcgtgcgata gttcgttttg
     8641 tcggtaatcc gatgttgccg cgccgatatc atgtgatgtt gtcacagtgc gcgaaattcg
     8701 aatggtggtg tgcagtgatt gtgttgtgac ggcgagtggc gcgtgtgggt gcttagtttt
     8761 gggagatgtt ttcgtatttt tttgttgata actcaggctt tgttgctgtg ttgtagtact
     8821 attttccatt gcgcggtgtc cagcttttaa ttagtggcac atattcttag caagtaaaaa
     8881 ttattttgca tactattaaa tttcttataa attattttct aaaattaagt ttaccttttc
     8941 aattttacta aaaatatcga tatatttatt atcgctggaa aactacatta ttccacctct
     9001 aagcaagaac cgttagttgg cgcgtagctt taccacaaaa ttcctggaat tgccgtacgc
     9061 ttcgcagttg tttcaagttg tctaagggac atacgatttt ttttgcctct gcgtcacgat
     9121 tttaacccaa aagcgagttt agttacatgt acattattat tagataaaga agtatcgcga
     9181 atacttcagt tgaataaact gtgcttggtt tttgggtgag gatttgtgga aagtagagtg
     9241 cgcgataacc gtaactttcg acccggattt tcgccactag ttctagagga tcgaaccctt
     9301 gccaccatgg acaagaagta ctccattggg ctcgatatcg gcacaaacag cgtcggctgg
     9361 gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc
     9421 gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggagacg
     9481 gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg
     9541 atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc
     9601 cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc
     9661 tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg
     9721 aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg
     9781 gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac
     9841 agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag
     9901 aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa
     9961 tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa cggcctgttt
    10021 ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa cttcgacctg
    10081 gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct cgacaatctg
    10141 ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa cctgtcagac
    10201 gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc tccgctgagc
    10261 gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct gaaggccctt
    10321 gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc taaaaatggc
    10381 tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt tattaagccc
    10441 atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag agaagatctg
    10501 ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca cctgggcgaa
    10561 ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga taacagggaa
    10621 aagattgaga aaatcctcac atttcggata ccctactatg taggccccct cgcccgggga
    10681 aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc ctggaacttc
    10741 gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat gactaacttt
    10801 gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta cgagtacttc
    10861 acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag aaagccagca
    10921 ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac gaaccggaaa
    10981 gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt cgactctgtt
    11041 gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca cgatctcctg
    11101 aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat tcttgaggac
    11161 attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg cttgaaaact
    11221 tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg atatacagga
    11281 tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag tggaaagaca
    11341 atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca gttgatccat
    11401 gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg ccagggggac
    11461 agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa gggaatactg
    11521 cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat
    11581 atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa gaacagtagg
    11641 gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat ccttaaggaa
    11701 cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta cctgcagaac
    11761 ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga ctacgacgtg
    11821 gatcatatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa agtgttgaca
    11881 agatccgata aaaatagagg gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa
    11941 atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg gaagttcgat
    12001 aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagccgg cttcatcaaa
    12061 aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct cgattcacgc
    12121 atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt tattactctg
    12181 aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt gagagagatc
    12241 aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac tgcacttatc
    12301 aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt gtacgatgtt
    12361 aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt
    12421 tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg agagattcgg
    12481 aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga caagggtagg
    12541 gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc
    12601 gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag
    12661 ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga ttctcctaca
    12721 gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa
    12781 agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga aaaaaacccc
    12841 atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat cattaagctt
    12901 cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc
    12961 gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt cttgtatctg
    13021 gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa gcagctgttc
    13081 gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga attctccaaa
    13141 agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa taagcacagg
    13201 gataagccca tcagggagca ggcagaaaac attatccact tgtttactct gaccaacttg
    13261 ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg gtacacctct
    13321 acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct ctatgaaaca
    13381 agaatcgacc tctctcagct cggtggagac agcagggctg accccaagaa gaagaggaag
    13441 gtgtgaaagg gttcgatccc taccggtgag agggcgaatc cagctctgga gcagaggctc
    13501 tggcagcttt tgcagcgttt atataacatg aaatatatat acgcattccg atcaaagctg
    13561 ggttaaccag atagatagat agtaacgttt aaatagcgcc tggcgcgttc gattttaaag
    13621 agatttagag cgttatcccg tgcctataga tcttatagta tagacaacga acgatcactc
    13681 aaatccaagt caataattca agaatttatg tctgtttctg tgaaagggaa actaattttg
    13741 ttaaagaaga cttacaatat cgtaatactt gttcaatcgt cgtggccgat agaaatatct
    13801 tacaatccga aagttgatga atggaattgg tctgcaactg gtcgccttca tttcgtaaaa
    13861 tgttcgcttg cggccgaaaa atttcgatat atctacaatt gatctacaat ctttactaaa
    13921 ttttgaaaaa ggaacacttt gaatttcgaa ctgtcaatcg tatcattaga atttaatcta
    13981 aatttaaatc ttgctaaagg aaatagcaag gaacactttc gtcgtcggct acgcattcat
    14041 tgtaaaattt taaattttga cattccgcac tttttgatag ataagcgaag agtattttta
    14101 ttacatgtat cgcaagtatt catttcaaca cacatatcta
//
