LOCUS       sequence-360676-       11088 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sp.Cas9 and gRNA yeast expression vector for HDR-based gRNA 
            introduction. Selection = LEU2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11088)
  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 11088)
  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: pBBK07 pCAS-Tyr-[gRNA: BsaI GFP dropout] (LEU2...
FEATURES             Location/Qualifiers
     source          1..11088
                     /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             2939..4033
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     terminator      4039..4236
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(4387..5178,5179..5247))
                     /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     5010..5029
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5248..5352)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6055..10158
                     /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             6055..10158
                     /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             10168..10188
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10204..10227
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      10231..10420
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(10477..11065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10557..10576)
                     /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 gtctgcccct aagaagatcg tcgttttgcc aggtgaccac gttggtcaag aaatcacagc
     3001 cgaagccatt aaggttctta aagctatttc tgatgttcgt tccaatgtca agttcgattt
     3061 cgaaaatcat ttaattggtg gtgctgctat cgatgctaca ggtgttccac ttccagatga
     3121 ggcgctggaa gcctccaaga aggctgatgc cgttttgtta ggtgctgtgg gtggtcctaa
     3181 atggggaacc ggtagtgtta gacctgaaca aggtttacta aaaatccgta aagaacttca
     3241 attgtacgcc aacttaagac catgtaactt tgcatccgac tctcttttag acttatctcc
     3301 aatcaagcca caatttgcta aaggtactga cttcgttgtt gtcagagaat tagtgggagg
     3361 tatttacttt ggtaagagaa aggaggacga tggtgatggt gtcgcttggg atagtgaaca
     3421 atacaccgtt ccagaagtgc aaagaatcac aagaatggcc gctttcatgg ccctacaaca
     3481 tgagccacca ttgcctattt ggtccttgga taaagctaat gttttggcct cttcaagatt
     3541 atggagaaaa actgtggagg aaaccatcaa gaacgaattt cctacattga aggttcaaca
     3601 tcaattgatt gattctgccg ccatgatcct agttaagaac ccaacccacc taaatggtat
     3661 tataatcacc agcaacatgt ttggtgatat catctccgat gaagcctccg ttatcccagg
     3721 ttccttgggt ttgttgccat ctgcgtcctt ggcctctttg ccagacaaga acaccgcatt
     3781 tggtttgtac gaaccatgcc acggttctgc tccagatttg ccaaagaata aggtcaaccc
     3841 tatcgccact atcttgtctg ctgcaatgat gttgaaattg tcattgaact tgcctgaaga
     3901 aggtaaggcc attgaagatg cagttaaaaa ggttttggat gcaggtatca gaactggtga
     3961 tttaggtggt tccaacagta ccaccgaagt cggtgatgct gtcgccgaag aagttaagaa
     4021 aatccttgct taatcagtac tgacaataaa aagattcttg ttttcaagaa cttgtcattt
     4081 gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt
     4141 ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca
     4201 tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata ctaacgccgc
     4261 catccagtgt cgaaaacgag ctctcgagaa cccttaatca aaaaggatct tcacctagat
     4321 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4381 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4441 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4501 tggccccagt gctgcaatga taccgcggga cccacgctca ccggctccag atttatcagc
     4561 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4621 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4681 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4741 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4801 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4861 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4921 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4981 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5041 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5101 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5161 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5221 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5281 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5341 aggggttcgt cgacagtcga acaagaagca ggcaaagttt agagcactgc ccctccgcac
     5401 tcaaaaaaga aaaaactagg aggaaaataa aattctcaac cacacaaaca cataaacaca
     5461 tacaaataca aatacaagct tatttacttg acatcgcgcg atcttccact attcagcgcc
     5521 gtccgccctc tctcgtgttt tttgtttacg cgacaactat gcgaaatccg gagcaacggg
     5581 caaccgtttg gggaaagacc acacccacgc gcgatcgcca tggcaacgag gtcgcacacg
     5641 ccccacaccc agacctccct gcgagcgggc atgggtacaa tgtccccgtt gccacagaca
     5701 ccacttcgta gcacagcgca gagcgtagcg tgttgttgct gctgacaaaa gaaaattttt
     5761 cttagcaaag caaaggaggg gaagcacggg cagatagcac cgtaccatac ccttggaaac
     5821 tcgaaatgaa cgaagcagga aatgagagaa tgagagtttt gtaggtatat atagcggtag
     5881 tgtttgcgcg ttaccatcat cttctggatc tatctattgt tcttttcctc atcactttcc
     5941 cctttttcgc tcttcttctt gtcttttatt tctttctttt ttttaattgt tccctcgatt
     6001 ggctatctac caaagaatcc aaacttaata cacgtattta tttgtccaat taccatggat
     6061 aagaaatact caataggctt agatatcggc acaaatagcg tcggatgggc ggtgatcact
     6121 gatgaatata aggttccgtc taaaaagttc aaggttctgg gaaatacaga ccgccacagt
     6181 atcaaaaaaa atcttatagg ggctctttta tttgacagtg gagagacagc ggaagcgact
     6241 cgactcaaac ggacagctcg tagaaggtat acacgtcgga agaatcgtat ttgttatcta
     6301 caggagattt tttcaaatga gatggcgaaa gtagatgata gtttctttca tcgacttgaa
     6361 gagtcttttt tggtggaaga agacaagaag catgaacgtc atcctatttt tggaaatata
     6421 gtagatgaag ttgcttatca tgagaaatat ccaactatct atcatctgcg aaaaaaattg
     6481 gtagattcta ctgataaagc ggatttgcgc ttaatctatt tggccttagc gcatatgatt
     6541 aagtttcgtg gtcatttttt gattgaggga gatttaaatc ctgataatag tgatgtggac
     6601 aaactattta tccagttggt acaaacctac aatcaattat ttgaagaaaa ccctattaac
     6661 gcaagtggag tagatgctaa agcgattctt tctgcacgat tgagtaaatc aagacgatta
     6721 gaaaatctca ttgctcagct ccccggtgag aagaaaaatg gcttatttgg gaatctcatt
     6781 gctttgtcat tgggtttgac ccctaatttt aaatcaaatt ttgatttggc agaagatgct
     6841 aaattacagc tttcaaaaga tacttacgat gatgatttag ataatttatt ggcgcaaatt
     6901 ggagatcaat atgctgattt gtttttggca gctaagaatt tatcagatgc tattttactt
     6961 tcagatatcc taagagtaaa tactgaaata actaaggctc ccctatcagc ttcaatgatt
     7021 aaacgctacg atgaacatca tcaagacttg actcttttaa aagctttagt tcgacaacaa
     7081 cttccagaaa agtataaaga aatctttttt gatcaatcaa aaaacggata tgcaggttat
     7141 attgatgggg gagctagcca agaagaattt tataaattta tcaaaccaat tttagaaaaa
     7201 atggatggta ctgaggaatt attggtgaaa ctaaatcgtg aagatttgct gcgcaagcaa
     7261 cggacctttg acaacggctc tattccccat caaattcact tgggtgagct gcatgctatt
     7321 ttgagaagac aagaagactt ttatccattt ttaaaagaca atcgtgagaa gattgaaaaa
     7381 atcttgactt ttcgaattcc ttattatgtt ggtccattgg cgcgtggcaa tagtcgtttt
     7441 gcatggatga ctcggaagtc tgaagaaaca attaccccat ggaattttga agaagttgtc
     7501 gataaaggtg cttcagctca atcatttatt gaacgcatga caaactttga taaaaatctt
     7561 ccaaatgaaa aagtactacc aaaacatagt ttgctttatg agtattttac ggtttataac
     7621 gaattgacaa aggtcaaata tgttactgaa ggaatgcgaa aaccagcatt tctttcaggt
     7681 gaacagaaga aagccattgt tgatttactc ttcaaaacaa atcgaaaagt aaccgttaag
     7741 caattaaaag aagattattt caaaaaaata gaatgttttg atagtgttga aatttcagga
     7801 gttgaagata gatttaatgc ttcattaggt acctaccatg atttgctaaa aattattaaa
     7861 gataaagatt ttttggataa tgaagaaaat gaagatatct tagaggatat tgttttaaca
     7921 ttgaccttat ttgaagatag ggagatgatt gaggaaagac ttaaaacata tgctcacctc
     7981 tttgatgata aggtgatgaa acagcttaaa cgtcgccgtt atactggttg gggacgtttg
     8041 tctcgaaaat tgattaatgg tattagggat aagcaatctg gcaaaacaat attagatttt
     8101 ttgaaatcag atggttttgc caatcgcaat tttatgcagc tgatccatga tgatagtttg
     8161 acatttaaag aagacattca aaaagcacaa gtgtctggac aaggcgatag tttacatgaa
     8221 catattgcaa atttagctgg tagccctgct attaaaaaag gtattttaca gactgtaaaa
     8281 gttgttgatg aattggtcaa agtaatgggg cggcataagc cagaaaatat cgttattgaa
     8341 atggcacgtg aaaatcagac aactcaaaag ggccagaaaa attcgcgaga gcgtatgaaa
     8401 cgaatcgaag aaggtatcaa agaattagga agtcagattc ttaaagagca tcctgttgaa
     8461 aatactcaat tgcaaaatga aaagctctat ctctattatc tccaaaatgg aagagacatg
     8521 tatgtggacc aagaattaga tattaatcgt ttaagtgatt atgatgtcga tcacattgtt
     8581 ccacaaagtt tccttaaaga cgattcaata gacaataagg tcttaacgcg ttctgataaa
     8641 aatcgtggta aatcggataa cgttccaagt gaagaagtag tcaaaaagat gaaaaactat
     8701 tggagacaac ttctaaacgc caagttaatc actcaacgta agtttgataa tttaacgaaa
     8761 gctgaacgtg gaggtttgag tgaacttgat aaagctggtt ttatcaaacg ccaattggtt
     8821 gaaactcgcc aaatcactaa gcatgtggca caaattttgg atagtcgcat gaatactaaa
     8881 tacgatgaaa atgataaact tattcgagag gttaaagtga ttaccttaaa atctaaatta
     8941 gtttctgact tccgaaaaga tttccaattc tataaagtac gtgagattaa caattaccat
     9001 catgcccatg atgcgtatct aaatgccgtc gttggaactg ctttgattaa gaaatatcca
     9061 aaacttgaat cggagtttgt ctatggtgat tataaagttt atgatgttcg taaaatgatt
     9121 gctaagtctg agcaagaaat aggcaaagca accgcaaaat atttctttta ctctaatatc
     9181 atgaacttct tcaaaacaga aattacactt gcaaatggag agattcgcaa acgccctcta
     9241 atcgaaacta atggggaaac tggagaaatt gtctgggata aagggcgaga ttttgccaca
     9301 gtgcgcaaag tattgtccat gccccaagtc aatattgtca agaaaacaga agtacagaca
     9361 ggcggattct ccaaggagtc aattttacca aaaagaaatt cggacaagct tattgctcgt
     9421 aaaaaagact gggatccaaa aaaatatggt ggttttgata gtccaacggt agcttattca
     9481 gtcctagtgg ttgctaaggt ggaaaaaggg aaatcgaaga agttaaaatc cgttaaagag
     9541 ttactaggga tcacaattat ggaaagaagt tcctttgaaa aaaatccgat tgacttttta
     9601 gaagctaaag gatataagga agttaaaaaa gacttaatca ttaaactacc taaatatagt
     9661 ctttttgagt tagaaaacgg tcgtaaacgg atgctggcta gtgccggaga attacaaaaa
     9721 ggaaatgagc tggctctgcc aagcaaatat gtgaattttt tatatttagc tagtcattat
     9781 gaaaagttga agggtagtcc agaagataac gaacaaaaac aattgtttgt ggagcagcat
     9841 aagcattatt tagatgagat tattgagcaa atcagtgaat tttctaagcg tgttatttta
     9901 gcagatgcca atttagataa agttcttagt gcatataaca aacatagaga caaaccaata
     9961 cgtgaacaag cagaaaatat tattcattta tttacgttga cgaatcttgg agctcccgct
    10021 gcttttaaat attttgatac aacaattgat cgtaaacgat atacgtctac aaaagaagtt
    10081 ttagatgcca ctcttatcca tcaatccatc actggtcttt atgaaacacg cattgatttg
    10141 agtcagctag gaggtgacgg tggagggcca aaaaagaaaa gaaaagttga agatgcttct
    10201 gctcatcatc accatcacca tcaccattaa atccgctcta accgaaaagg aaggagttag
    10261 acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt
    10321 atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat
    10381 actgaaaacc ttgcttgaga aggttttggg acgctcgaag tgagcaaaag gccagcaaaa
    10441 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    10501 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    10561 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    10621 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    10681 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    10741 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    10801 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    10861 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
    10921 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    10981 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    11041 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctac
//
