LOCUS       sequence-360678-       10809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sp.Cas9 and gRNA yeast expression vector for HDR-based gRNA 
            introduction. Selection = URA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10809)
  AUTHORS   Bean BDM, Whiteway M, Martin VJJ
  TITLE     The MyLo CRISPR-Cas9 Toolkit: A Markerless Yeast Localization and 
            Overexpression CRISPR-Cas9 Toolkit.
  JOURNAL   G3 (Bethesda). 2022 Jun 16. pii: 6609175. doi: 
            10.1093/g3journal/jkac154.
  PUBMED    35708612
REFERENCE   2  (bases 1 to 10809)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "G3 
            (Bethesda). 2022 Jun 16. pii: 6609175. doi: 
            10.1093/g3journal/jkac154."
COMMENT     Sequence Label: pBBK09 pCAS-Tyr-[gRNA: BsaI GFP dropout]
            (BsmB...
FEATURES             Location/Qualifiers
     source          1..10809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(962..1009)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     RBS             1515..1526
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1533..2246
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2258..2329
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2345..2372
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        2386..2461
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2595..2937
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2948..3745
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="SKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELLE
                     LVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQY
                     SAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGEY
                     TKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDDV
                     VSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     terminator      3760..3957
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(4108..4899,4900..4968))
                     /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     4731..4750
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4969..5073)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5776..9879
                     /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             5776..9879
                     /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             9889..9909
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9925..9948
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      9952..10141
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(10198..10786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10278..10297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca
       61 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta
      121 ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt
      181 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta
      241 ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc
      301 tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca
      361 gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg
      421 tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc
      481 gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt
      541 ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa
      601 aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt
      661 ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga
      721 gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag
      781 cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag caaagagata
      841 cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagcccg ttacagtccg
      901 gtgcgttttt ggttttttga aagtgcgtct tcagagcgct tttggttttc aaaagcgctc
      961 tgaagttcct atactttcta gagaatagga acttcggaat aggaacttca aagcgtttcc
     1021 gaaaacgagc gcttccgaaa atgcaacgcg agctgcgcac atacagctca ctgttcacgt
     1081 cgcacctata tctgcgtgtt gcctgtatat atatatacat gagaagaacg gcatagtgcg
     1141 tgtttatgct taaatgcgta tatgtgttat gtagtatact ctttcttcaa caattaaata
     1201 ctctcggtag ccaagttggt ttaaggcgca agactgtaat ttatcactac gaaatcttga
     1261 gatcgggcgt tcgactcgcc cccgggagag atggccggca tggtcccagc ctcctcgctg
     1321 gcgccggctg ggcaacacct tcgggtggcg aatgggactc gagaccgaaa gtgaaacgtg
     1381 atttcatgcg tcattttgaa cattttgtaa atcttattta ataatgtgtg cggcaattca
     1441 catttaattt atgaatgttt tcttaacatc gcggcaactc aagaaacggc aggttcggat
     1501 cttagctact agagaaagag gagaaatact agatgcgtaa aggcgaagag ctgttcactg
     1561 gtgtcgtccc tattctggtg gaactggatg gtgatgtcaa cggtcataag ttttccgtgc
     1621 gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac gctgaagttc atctgtacta
     1681 ctggtaaact gccggttcct tggccgactc tggtaacgac gctgacttat ggtgttcagt
     1741 gctttgctcg ttatccggac catatgaagc agcatgactt cttcaagtcc gccatgccgg
     1801 aaggctatgt gcaggaacgc acgatttcct ttaaggatga cggcacgtac aaaacgcgtg
     1861 cggaagtgaa atttgaaggc gataccctgg taaaccgcat tgagctgaaa ggcattgact
     1921 ttaaagagga cggcaatatc ctgggccata agctggaata caattttaac agccacaatg
     1981 tttacatcac cgccgataaa caaaaaaatg gcattaaagc gaattttaaa attcgccaca
     2041 acgtggagga tggcagcgtg cagctggctg atcactacca gcaaaacact ccaatcggtg
     2101 atggtcctgt tctgctgcca gacaatcact atctgagcac gcaaagcgtt ctgtctaaag
     2161 atccgaacga gaaacgcgat catatggttc tgctggagtt cgtaaccgca gcgggcatca
     2221 cgcatggtat ggatgaactg tacaaatgac caggcatcaa ataaaacgaa aggctcagtc
     2281 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
     2341 cactggctca ccttcgggtg ggcctttctg cgtttatagg tctccgtttt agagctagaa
     2401 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     2461 ctttttttat tttttgtcac tattgttatg taaaatgcca cctctgacag tatggaacgc
     2521 aaacttctgt ctagtggata taggtctaga gatctgttta gcttgcctcg tccccgccgg
     2581 gtcacccggc cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca
     2641 gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat
     2701 catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc
     2761 tgcagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg
     2821 cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac
     2881 ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaacaat
     2941 ggagacgtcg aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc
     3001 caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg
     3061 taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa
     3121 aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc
     3181 attatccgcc aagtacaatt ttttactctt cgaggacaga aaatttgctg acattggtaa
     3241 tacagtcaaa ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac
     3301 gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga
     3361 agtaacaaag gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct
     3421 atctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt
     3481 tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat
     3541 tatgacaccc ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac agtatagaac
     3601 cgtggatgat gtggtttcta caggatctga cattattatt gttggaagag gactatttgc
     3661 aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata
     3721 tttgagaaga tgcggccagc aaaaccgtct ctaatcagta ctgacaataa aaagattctt
     3781 gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca
     3841 aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta
     3901 agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg
     3961 tcgattcgat actaacgccg ccatccagtg tcgaaaacga gctctcgaga acccttaatc
     4021 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4081 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4141 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4201 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggg acccacgctc
     4261 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4321 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4381 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4441 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4501 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4561 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4621 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4681 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4741 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4801 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4861 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4921 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4981 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5041 tatttagaaa aataaacaaa taggggttcg tcgacagtcg aacaagaagc aggcaaagtt
     5101 tagagcactg cccctccgca ctcaaaaaag aaaaaactag gaggaaaata aaattctcaa
     5161 ccacacaaac acataaacac atacaaatac aaatacaagc ttatttactt gacatcgcgc
     5221 gatcttccac tattcagcgc cgtccgccct ctctcgtgtt ttttgtttac gcgacaacta
     5281 tgcgaaatcc ggagcaacgg gcaaccgttt ggggaaagac cacacccacg cgcgatcgcc
     5341 atggcaacga ggtcgcacac gccccacacc cagacctccc tgcgagcggg catgggtaca
     5401 atgtccccgt tgccacagac accacttcgt agcacagcgc agagcgtagc gtgttgttgc
     5461 tgctgacaaa agaaaatttt tcttagcaaa gcaaaggagg ggaagcacgg gcagatagca
     5521 ccgtaccata cccttggaaa ctcgaaatga acgaagcagg aaatgagaga atgagagttt
     5581 tgtaggtata tatagcggta gtgtttgcgc gttaccatca tcttctggat ctatctattg
     5641 ttcttttcct catcactttc ccctttttcg ctcttcttct tgtcttttat ttctttcttt
     5701 tttttaattg ttccctcgat tggctatcta ccaaagaatc caaacttaat acacgtattt
     5761 atttgtccaa ttaccatgga taagaaatac tcaataggct tagatatcgg cacaaatagc
     5821 gtcggatggg cggtgatcac tgatgaatat aaggttccgt ctaaaaagtt caaggttctg
     5881 ggaaatacag accgccacag tatcaaaaaa aatcttatag gggctctttt atttgacagt
     5941 ggagagacag cggaagcgac tcgactcaaa cggacagctc gtagaaggta tacacgtcgg
     6001 aagaatcgta tttgttatct acaggagatt ttttcaaatg agatggcgaa agtagatgat
     6061 agtttctttc atcgacttga agagtctttt ttggtggaag aagacaagaa gcatgaacgt
     6121 catcctattt ttggaaatat agtagatgaa gttgcttatc atgagaaata tccaactatc
     6181 tatcatctgc gaaaaaaatt ggtagattct actgataaag cggatttgcg cttaatctat
     6241 ttggccttag cgcatatgat taagtttcgt ggtcattttt tgattgaggg agatttaaat
     6301 cctgataata gtgatgtgga caaactattt atccagttgg tacaaaccta caatcaatta
     6361 tttgaagaaa accctattaa cgcaagtgga gtagatgcta aagcgattct ttctgcacga
     6421 ttgagtaaat caagacgatt agaaaatctc attgctcagc tccccggtga gaagaaaaat
     6481 ggcttatttg ggaatctcat tgctttgtca ttgggtttga cccctaattt taaatcaaat
     6541 tttgatttgg cagaagatgc taaattacag ctttcaaaag atacttacga tgatgattta
     6601 gataatttat tggcgcaaat tggagatcaa tatgctgatt tgtttttggc agctaagaat
     6661 ttatcagatg ctattttact ttcagatatc ctaagagtaa atactgaaat aactaaggct
     6721 cccctatcag cttcaatgat taaacgctac gatgaacatc atcaagactt gactctttta
     6781 aaagctttag ttcgacaaca acttccagaa aagtataaag aaatcttttt tgatcaatca
     6841 aaaaacggat atgcaggtta tattgatggg ggagctagcc aagaagaatt ttataaattt
     6901 atcaaaccaa ttttagaaaa aatggatggt actgaggaat tattggtgaa actaaatcgt
     6961 gaagatttgc tgcgcaagca acggaccttt gacaacggct ctattcccca tcaaattcac
     7021 ttgggtgagc tgcatgctat tttgagaaga caagaagact tttatccatt tttaaaagac
     7081 aatcgtgaga agattgaaaa aatcttgact tttcgaattc cttattatgt tggtccattg
     7141 gcgcgtggca atagtcgttt tgcatggatg actcggaagt ctgaagaaac aattacccca
     7201 tggaattttg aagaagttgt cgataaaggt gcttcagctc aatcatttat tgaacgcatg
     7261 acaaactttg ataaaaatct tccaaatgaa aaagtactac caaaacatag tttgctttat
     7321 gagtatttta cggtttataa cgaattgaca aaggtcaaat atgttactga aggaatgcga
     7381 aaaccagcat ttctttcagg tgaacagaag aaagccattg ttgatttact cttcaaaaca
     7441 aatcgaaaag taaccgttaa gcaattaaaa gaagattatt tcaaaaaaat agaatgtttt
     7501 gatagtgttg aaatttcagg agttgaagat agatttaatg cttcattagg tacctaccat
     7561 gatttgctaa aaattattaa agataaagat tttttggata atgaagaaaa tgaagatatc
     7621 ttagaggata ttgttttaac attgacctta tttgaagata gggagatgat tgaggaaaga
     7681 cttaaaacat atgctcacct ctttgatgat aaggtgatga aacagcttaa acgtcgccgt
     7741 tatactggtt ggggacgttt gtctcgaaaa ttgattaatg gtattaggga taagcaatct
     7801 ggcaaaacaa tattagattt tttgaaatca gatggttttg ccaatcgcaa ttttatgcag
     7861 ctgatccatg atgatagttt gacatttaaa gaagacattc aaaaagcaca agtgtctgga
     7921 caaggcgata gtttacatga acatattgca aatttagctg gtagccctgc tattaaaaaa
     7981 ggtattttac agactgtaaa agttgttgat gaattggtca aagtaatggg gcggcataag
     8041 ccagaaaata tcgttattga aatggcacgt gaaaatcaga caactcaaaa gggccagaaa
     8101 aattcgcgag agcgtatgaa acgaatcgaa gaaggtatca aagaattagg aagtcagatt
     8161 cttaaagagc atcctgttga aaatactcaa ttgcaaaatg aaaagctcta tctctattat
     8221 ctccaaaatg gaagagacat gtatgtggac caagaattag atattaatcg tttaagtgat
     8281 tatgatgtcg atcacattgt tccacaaagt ttccttaaag acgattcaat agacaataag
     8341 gtcttaacgc gttctgataa aaatcgtggt aaatcggata acgttccaag tgaagaagta
     8401 gtcaaaaaga tgaaaaacta ttggagacaa cttctaaacg ccaagttaat cactcaacgt
     8461 aagtttgata atttaacgaa agctgaacgt ggaggtttga gtgaacttga taaagctggt
     8521 tttatcaaac gccaattggt tgaaactcgc caaatcacta agcatgtggc acaaattttg
     8581 gatagtcgca tgaatactaa atacgatgaa aatgataaac ttattcgaga ggttaaagtg
     8641 attaccttaa aatctaaatt agtttctgac ttccgaaaag atttccaatt ctataaagta
     8701 cgtgagatta acaattacca tcatgcccat gatgcgtatc taaatgccgt cgttggaact
     8761 gctttgatta agaaatatcc aaaacttgaa tcggagtttg tctatggtga ttataaagtt
     8821 tatgatgttc gtaaaatgat tgctaagtct gagcaagaaa taggcaaagc aaccgcaaaa
     8881 tatttctttt actctaatat catgaacttc ttcaaaacag aaattacact tgcaaatgga
     8941 gagattcgca aacgccctct aatcgaaact aatggggaaa ctggagaaat tgtctgggat
     9001 aaagggcgag attttgccac agtgcgcaaa gtattgtcca tgccccaagt caatattgtc
     9061 aagaaaacag aagtacagac aggcggattc tccaaggagt caattttacc aaaaagaaat
     9121 tcggacaagc ttattgctcg taaaaaagac tgggatccaa aaaaatatgg tggttttgat
     9181 agtccaacgg tagcttattc agtcctagtg gttgctaagg tggaaaaagg gaaatcgaag
     9241 aagttaaaat ccgttaaaga gttactaggg atcacaatta tggaaagaag ttcctttgaa
     9301 aaaaatccga ttgacttttt agaagctaaa ggatataagg aagttaaaaa agacttaatc
     9361 attaaactac ctaaatatag tctttttgag ttagaaaacg gtcgtaaacg gatgctggct
     9421 agtgccggag aattacaaaa aggaaatgag ctggctctgc caagcaaata tgtgaatttt
     9481 ttatatttag ctagtcatta tgaaaagttg aagggtagtc cagaagataa cgaacaaaaa
     9541 caattgtttg tggagcagca taagcattat ttagatgaga ttattgagca aatcagtgaa
     9601 ttttctaagc gtgttatttt agcagatgcc aatttagata aagttcttag tgcatataac
     9661 aaacatagag acaaaccaat acgtgaacaa gcagaaaata ttattcattt atttacgttg
     9721 acgaatcttg gagctcccgc tgcttttaaa tattttgata caacaattga tcgtaaacga
     9781 tatacgtcta caaaagaagt tttagatgcc actcttatcc atcaatccat cactggtctt
     9841 tatgaaacac gcattgattt gagtcagcta ggaggtgacg gtggagggcc aaaaaagaaa
     9901 agaaaagttg aagatgcttc tgctcatcat caccatcacc atcaccatta aatccgctct
     9961 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt tttttatagt
    10021 tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacagacg
    10081 cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg gacgctcgaa
    10141 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
    10201 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
    10261 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
    10321 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
    10381 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
    10441 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
    10501 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
    10561 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
    10621 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
    10681 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
    10741 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
    10801 cttttctac
//
