LOCUS       sequence-331098-       15602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 in Drosophila late germline cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15602)
  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 15602)
  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: BicC.Ca9.R
FEATURES             Location/Qualifiers
     source          1..15602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..82
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     276..298
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     290..307
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     291..307
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   1011..1073
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    2107..2176
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     CDS             5221..5286
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5293..5313
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5338..9438
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5338..9438
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9439..9486
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9487..9540
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(9541..9543,9544..9546,9547..10260)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTSKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9847..9866)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     10194..10215
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             join(11495..11497,11498..11500,11501..12175)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(11678..11698)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     11896..11915
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     12090..12113
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    12289..12410
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12326..12345)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12380..12399
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   13078..13112
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(13293..13309)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13293..13309)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13306..13328)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13317..13333
                     /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(13341..13347,13348..13365,13366..13371))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13386..13407
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13524..13541)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13695..14283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13775..13794)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14454..15245,15246..15314))
                     /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     15077..15096
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15315..15419)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     15487..15505
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(15543..15565)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
       61 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
      121 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
      181 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
      241 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
      301 ggccagtgcc aagctttgtt taaaaatata acaaaattgt gatcccacaa aatgaagtgg
      361 ggcaaaatca aataattaat agtgtccgta aacttgttgg tcttcaactt tttgaggaac
      421 acgttggacg gcaaatccgt gactataaca caagttgatt taataatttt agccaacacg
      481 tcgggctgcg tgttttttgc cgacgcgtct gtgtacacgt tgattaactg gtcgattaaa
      541 ctgttgaaat aatttaattt ttggttcttc tttaaatctg tgatgaaatt ttttaaaata
      601 actttaaatt cttcattggt aaaaaatgcc acgttttgca acttgtgagg gtctaatatg
      661 aggtcaaact cagtaggagt tttatccaaa aaagaaaaca tgattacgtc tgtacacgaa
      721 cgcgtattaa cgcagagtgc aaagtataag agggttaaaa aatatatttt acgcaccata
      781 tacgcatcgg gttgatatcg ttaatatgga tcaatttgaa cagttgatta acgtgtctct
      841 gctcaagtct ttgatcaaaa cgcaaatcga cgaaaatgtg tcggacaata tcaagtcgat
      901 gagcgaaaaa ctaaaaaggc tagaatacga caatctcaca gacagcgttg agatatacgg
      961 tattcacgac agcaggctga ataataaaaa aattagaaac tattatttaa ccctagaaag
     1021 ataatcatat tgtgacgtac gttaaagata atcatgcgta aaattgacgc atgtgtttta
     1081 tcggtctgta tatcgaggtt tatttattaa tttgaataga tattaagttt tattatattt
     1141 acacttacat actaataata aattcaacaa acaatttatt tatgtttatt tatttattaa
     1201 aaaaaaacaa aaactcaaaa tttcttctat aaagtaacaa aacttttaaa cattctctct
     1261 tttacaaaaa taaacttatt ttgtacttta aaaacagtca tgttgtatta taaaataagt
     1321 aattagctta acttatacat aatagaaaca aattatactt attagtcagt cagaaacaac
     1381 tttggcacat atcaatatta tgctctcgac aaataacttt tttgcatttt ttgcacgatg
     1441 catttgcctt tcgccttatt ttagaggggc agtaagtaca gtaagtacgt tttttcatta
     1501 ctggctcttc agtactgtca tctgatgtac caggcacttc atttggcaaa atattagaga
     1561 tattatcgcg caaatatctc ttcaaagtag gagcttctaa acgcttacgc ataaacgatg
     1621 acgtcaggct catgtaaagg tttctcataa attttttgcg actttgaacc ttttctccct
     1681 tgctactgac attatggctg tatataataa aagaatttat gcaggcaatg tttatcattc
     1741 cgtacaataa tgccataggc cacctattcg tcttcctact gcaggtcatc acagaacaca
     1801 tttggtctag cgtgtccact ccgcctttag tttgattata atacataacc atttgcggtt
     1861 taccggtact ttcgttgata gaagcatcct catcacaaga tgataataag tataccatct
     1921 tagctggctt cggtttatat gagacgagag taaggggtcc gtcaaaacaa aacatcgatg
     1981 ttcccactgg cctggagcga ctgtttttca gtacttccgg tatctcgcgt ttgtttgatc
     2041 gcacggttcc cacaatggtt aattcgagct cgcccggggt cctaggtcga cgatgtaggt
     2101 cacggtctcg aagccgcggt gcgggtgcca gggcgtgccc ttgggctccc cgggcgcgta
     2161 ctccacctca cccatctggt ccatcatgat gaacgggtcg aggtggcggt agttgatccc
     2221 ggcgaacgcg cggcgcaccg ggaagccctc gccctcgaaa ccgctgggcg cggtggtcac
     2281 ggtgagcacg ggacgtgcga cggcgtcggc gggtgcggat acgcggggca gcgtcagcgg
     2341 gttctcgacg gtcacggcgg gcatgtcgac gcggccgcat aattatataa taataaactg
     2401 catgccgcca gatttttgta atcaaatcgt gttgtgaaag ttgtcttccc taatttatgc
     2461 ccattacatt aaaacattga caggggattt taattttaat ttcagtatgg tttcaattat
     2521 ggccaatata tataataaac tataaccata tgtataagaa tgtaaaaagc tagttttatg
     2581 tacataaaaa tgggacaaac attttgatgt tactgaataa atattaaaag tattaaccaa
     2641 gaaatatgaa agtaggataa aaaaacagtg aatgtttcca gctgctagga aaaacgataa
     2701 gtcttagatc tataaagaga atcaaaaacg cggtcaaatg catagcacat aatatattca
     2761 tatccccaat aaataacccg aaatgtgtaa agccagaata aatgtaataa taagattgaa
     2821 gaaaacaaaa tagtgatgta aaatagcagc aaggaaataa agtaacccct attcaataac
     2881 tattgctgac gtcttttact cgtacatgtc atcatttaat tacgtaattt aagtttggtt
     2941 tatttaaagt ttgatttccg gttgtacaag ggtgagcgta cttttaaaac ccactgtata
     3001 taaatacttt ggtaaagtgc aggttgggga gtttcagaag cttgaaaagc tccgtttccc
     3061 gttataaagc tattgctgaa tatccgttat ccagttggct ggcagctttt taaggttgtt
     3121 cgtgcactgg cttcactccc cacagaacac agagcttttg gatctgaagg tcagttggga
     3181 ctgggaccca catgccgagc ttagtttgat agtcagcgcg ctttaatgac gacaactgca
     3241 cggcgcgcac aacccattaa taatatatta attcaaaaga gctataaaat attaaaaaat
     3301 attcaaatat aactaactaa cgcgcagtgc acgattttct gacaacaatt cagtttcgct
     3361 tttcatgaga ctccattgca ttttcctttg tgtttttcac acacgtggtg tttttctgtt
     3421 actcgcactt cgatagtggt tatttttttt ggttacattg aaaaaataca ccgactgcca
     3481 gccgaattcc cattcgaacc ttaatttgac caattgacgg aaaattaatt gcgcccaaaa
     3541 ttaattagca agcggatata caattttttt acgtgaaaag agcgtgttat ttgtcgaaaa
     3601 gattttacaa ttgattgtgt ttaagtaaat taacctgcga tatctatgtg tatgtgctac
     3661 gatccgaatt tctgacatta cggtaagtgg cgtgtttttg gcgggaataa agtcggtttc
     3721 tctttgtgcc gtatttctct accgtttttt ctggcgccaa aatgaatgaa aacggttggc
     3781 aacatttcaa acttaacatt gcaacattct aaccgctgtt aagcgtttgc tctgcaaatt
     3841 gttgtagcta tgttggagtt ctggaacacc acctctccct gcgactttgt tttgtttttc
     3901 gctctcgaca tatggaatcg aatctcttat tccgggagcg gttcattgac ccagcaacaa
     3961 caatattcgc cagattttct cctgccagcg atttgcggaa ttcatccgct caacagttag
     4021 aagtaaatgc cgtcgagcag cgggcaaaat cgatcgacga aatcgagagc cgcgttcagt
     4081 tgccttcaag ccgtacttag aagaagtgct ggtcccccag aaatcagaga gatttttccc
     4141 aacttgcatt tttccaatac tctgacaaag cccctcagac cgtcgcaatc acacagctat
     4201 atactgatat atagatctct attgcgcaga tcacacctta aatgcctttc gtcgagagat
     4261 ttcaacaaat acgtaatttt gttatgaacg tctgcctcgt cgaggccatc caattcgtcc
     4321 aggacctctc gcacgccatc tgtctggacc tccagggcct atttaatctc aagtagagat
     4381 caccgagcga tctcccagcg gtaagttacg ccattggttt ttgggcacaa ggaagtgaaa
     4441 gtgaagacac gggtgaaatt acggttttcg ggatctcgag agaaagggga aagtacgaac
     4501 ctcatggcag cagcactcca ttttgagagg ttatgtgaaa gaacacacaa gaaagtttgg
     4561 aattttcgac aggcggcgaa ccaataataa tgttgttatc cataataata ttaattagtg
     4621 aatttttatt tcaccttttc gcggcattat tctcacaaat ttataagaca gattttcaaa
     4681 ttaagacaga aagtttttta atgttagtga aaagtactta ttccagttga tagttaaata
     4741 ctatttaccg caaaaaatta aaagagaact aaatttaaag gtaaaatcgt tttttttttt
     4801 ggaaagtgca ttgcttaaat tatggtaccg actatcaaaa taaaaataat taatttttat
     4861 atggtgtttc aatattcact ttaaattacc tatgactcgc tttctaacca ctctcttctt
     4921 attacgaaca tcaattttct tgttacatca tttcggacct ttcatttttc atcaccagct
     4981 catcccatca attcagatac actttacttc attaagtagc agcaatcatc taacaaagct
     5041 gaccaaacaa cggaccggct tccagaatcc ttaactgaga cggacgtttt gcttggtttt
     5101 taaaactaaa cgactttttg gtacccacac ctgttgacct ctgacgaatt tctcgatcca
     5161 ctgaccgatc cactgatcca gtgatcatca tcatcatccg aattccacag tttaaacatg
     5221 gactataagg accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac
     5281 gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac
     5341 aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc
     5401 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
     5461 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
     5521 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
     5581 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
     5641 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
     5701 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
     5761 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
     5821 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
     5881 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
     5941 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
     6001 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt
     6061 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
     6121 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
     6181 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     6241 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     6301 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     6361 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     6421 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     6481 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     6541 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     6601 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     6661 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     6721 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     6781 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     6841 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     6901 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     6961 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     7021 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     7081 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     7141 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     7201 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     7261 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     7321 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     7381 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     7441 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
     7501 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     7561 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa
     7621 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     7681 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     7741 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     7801 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg
     7861 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     7921 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     7981 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     8041 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     8101 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     8161 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     8221 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     8281 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     8341 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc
     8401 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc
     8461 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     8521 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     8581 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     8641 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga
     8701 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct
     8761 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     8821 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     8881 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     8941 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag
     9001 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     9061 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     9121 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     9181 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     9241 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     9301 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg
     9361 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg
     9421 tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag
     9481 aaaaaggagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct
     9541 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     9601 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     9661 ggcaagctga ccctgaagtt catctgcacc accagcaagc tgcccgtgcc ctggcccacc
     9721 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     9781 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     9841 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     9901 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     9961 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
    10021 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
    10081 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
    10141 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
    10201 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
    10261 ttaattaact agaatgaatc gtttttaaaa taacaaatca attgttttat aatattcgta
    10321 cgattctttg attatgtaat aaaatgtgat cattaggaag attacgaaaa atataaaaaa
    10381 tatgagttct gtgtgtataa caaatgctgt aaacgccaca attgtgtttg ttgcaaataa
    10441 acccatgatt atttgattaa aattgttgtt ttctttgttc atagacaata gtgtgttttg
    10501 cctaaacgtg tactgcataa actccatgcg agtgtatagc gagctagtgg ctaacgcttg
    10561 ccccaccaaa gtagattcgt caaaatcctc aatttcatca ccctcctcca agtttaacat
    10621 ttggccgtcg gaattaactt ctaaagatgc cacataatct aataaatgaa atagagattc
    10681 aaacgtggcg tcatcgtccg tttcgaccat ttccgaaaag aactcgggca taaactctat
    10741 gatttctctg gacgtggtgt tgtcgaaact ctcaaagtac gcagtcagga acgtgcgcga
    10801 catgtcgtcg ggaaactcgc gcggaaacat gttgttgtaa ccgaacgggt cccatagcgc
    10861 caaaaccaaa tctgccagcg tcaatagaat gagcacgatg ccgacaatgg agctggcttg
    10921 gatagcgatt cgagttaacg gccggggcgc gcccatactc ggtggcctcc ccaccaccaa
    10981 cttttttgca ctgcaaaaaa acacgctttt gcacgcgggc ccatacatag tacaaactct
    11041 acgtttcgta gactatttta cataaatagt ctacaccgtt gtatacgctc caaatacact
    11101 accacacatt gaaccttttt gcagtgcaaa aaagtacgtg tcggcagtca cgtaggccgg
    11161 ccttatcggg tcgcgtcctg tcacgtacga atcacattat cggaccggac gagtgttgtc
    11221 ttatcgtgac aggacgccag cttcctgtgt tgctaaccgc agccggacgc aactccttat
    11281 cggaacagga cgcgcctcca tatcagccgc gcgttatctc atgcgcgtga ccggacacga
    11341 ggcgcccgtc ccgcttatcg cgcctataaa tacagcccgc aacgatctgg taaacacagt
    11401 tgaacagcat ctgttacagc gacacaacat gagccggtcc aacaacgcca acgcgcccac
    11461 gccatccaac cgccgccgca acctgtctct ggtgatggtg cgctcctcca agaacgtcat
    11521 caaggagttc atgcgcttca aggtgcgcat ggagggcacc gtgaacggcc acgagttcga
    11581 gatcgagggc gagggcgagg gccgccccta cgagggccac aacaccgtga agctgaaggt
    11641 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt tccagtacgg
    11701 ctccaaggtg tacgtgaagc accccgccga catccccgac tacaagaagc tgtccttccc
    11761 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
    11821 ccaggactcc tccctgcagg acggctgctt catctacaag gtgaagttca tcggcgtgaa
    11881 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga
    11941 gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc cacaaggccc tgaagctgaa
    12001 ggacggcggc cactacctgg tggagttcaa gtccatctac atggccaaga agcccgtgca
    12061 gctgcccggc tactactacg tggactccaa gctggacatc acctcccaca acgaggacta
    12121 caccatcgtg gagcagtacg agcgcaccga gggccgccac cacctgttcc tgtaggactc
    12181 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
    12241 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
    12301 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
    12361 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aagcttctag
    12421 gccggccgat ctcggatctg acaatgttca gtgcagagac tcggctacgc ctcgtggact
    12481 ttgaagttga ccaacaatgt ttattcttac ctctaatagt cctctgtggc aaggtcaaga
    12541 ttctgttaga agccaatgaa gaacctggtt gttcaataac attttgttcg tctaatattt
    12601 cactaccgct tgacgttggc tgcacttcat gtacctcatc tataaacgct tcttctgtat
    12661 cgctctggac gtcatcttca cttacgtgat ctgatatttc actgtcagaa tcctcaccaa
    12721 caagctcgtc atcgctttgc agaagagcag agaggatatg ctcatcgtct aaagaactac
    12781 ccattttatt atatattagt cacgatatct ataacaagaa aatatatata taataagtta
    12841 tcacgtaagt agaacatgaa ataacaatat aattatcgta tgagttaaat cttaaaagtc
    12901 acgtaaaaga taatcatgcg tcattttgac tcacgcggtc gttatagttc aaaatcagtg
    12961 acacttaccg cattgacaag cacgcctcac gggagctcca agcggcgact gagatgtcct
    13021 aaatgcacag cgacggattc gcgctattta gaaagagaga gcaatatttc aagaatgcat
    13081 gcgtcaattt tacgcagact atctttctag ggttaaaaaa gatttgcgct ttactcgacc
    13141 taaactttaa acacgtcata gaatcttcgt ttgacaaaaa ccacattgtg gccaagctgt
    13201 gtgacgcgac gcgcgctaaa gaatggcaaa ccaagtcgcg cgagcgtcga ctctagagga
    13261 tccccgggta ccgagctcga attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt
    13321 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
    13381 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
    13441 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
    13501 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
    13561 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
    13621 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
    13681 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
    13741 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
    13801 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
    13861 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
    13921 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
    13981 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
    14041 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
    14101 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
    14161 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
    14221 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
    14281 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
    14341 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
    14401 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
    14461 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
    14521 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
    14581 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
    14641 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
    14701 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
    14761 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
    14821 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
    14881 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
    14941 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
    15001 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
    15061 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
    15121 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
    15181 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
    15241 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
    15301 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
    15361 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
    15421 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
    15481 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
    15541 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
    15601 ag
//
