LOCUS       sequence-360675-       10885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sp.Cas9 and gRNA yeast expression vector for Golden Gate pre-cloning
            of gRNA. Selection = SplitNourseothrycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10885)
  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 10885)
  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: pBBK06 pCAS-Tyr-[gRNA: BsaI GFP dropout]
            (Spli...
FEATURES             Location/Qualifiers
     source          1..10885
                     /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"
     terminator      3836..4033
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(4184..4975,4976..5044))
                     /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     4807..4826
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5045..5149)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5852..9955
                     /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             5852..9955
                     /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             9965..9985
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10001..10024
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      10028..10217
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(10274..10862)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10354..10373)
                     /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 gggtactacc ttagatgata cagcctacag atacagaaca tcagtccctg gtgatgctga
     3001 agcaattgag gctttagacg gttcattcac caccgacacc gtctttagag taaccgccac
     3061 cggtgatgga tttaccttaa gagaagtccc agtcgaccct ccattaacta aagtctttcc
     3121 agatgatgaa tctgatgacg aaagcgacga cggagaagat ggtgacccag attcaagaac
     3181 tttcgtagca tacggtgatg acggtgattt ggctggtttt gtagtcgttt cttattcagg
     3241 ttggaataga aggttgaccg ttgaagatat agaagtcgcc ccagagcata gaggtcatgg
     3301 tgtaggaaga gctttgatgg gtttggctac agaatttgca agagagagag gagccggtca
     3361 tttatggttg acgtcgctgt cctgcaggat ttaccttaag agaagtccca gtcgaccctc
     3421 cattaactaa agtctttcca gatgatgaat ctgatgacga aagcgacgac ggagaagatg
     3481 gtgacccaga ttcaagaact ttcgtagcat acggtgatga cggtgatttg gctggttttg
     3541 tagtcgtttc ttattcaggt tggaatagaa ggttgaccgt tgaagatata gaagtcgccc
     3601 cagagcatag aggtcatggt gtaggaagag ctttgatggg tttggctaca gaatttgcaa
     3661 gagagagagg agccggtcat ttatggttag aagttactaa tgttaacgcc cctgctatcc
     3721 atgcttatag aagaatgggt ttcacattat gtggtttaga tactgcttta tatgatggaa
     3781 cagcatctga cggtgaacag gccttgtata tgtctatgcc ttgcccttaa tcagtactga
     3841 caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt atattgtagt
     3901 tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg acatcatctg
     3961 cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct
     4021 ggtcgctata ctgctgtcga ttcgatacta acgccgccat ccagtgtcga aaacgagctc
     4081 tcgagaaccc ttaatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4141 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4201 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4261 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4321 cgcgggaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4381 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4441 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4501 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4561 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4621 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4681 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4741 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4801 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4861 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4921 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4981 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5041 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5101 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttcgtcga cagtcgaaca
     5161 agaagcaggc aaagtttaga gcactgcccc tccgcactca aaaaagaaaa aactaggagg
     5221 aaaataaaat tctcaaccac acaaacacat aaacacatac aaatacaaat acaagcttat
     5281 ttacttgaca tcgcgcgatc ttccactatt cagcgccgtc cgccctctct cgtgtttttt
     5341 gtttacgcga caactatgcg aaatccggag caacgggcaa ccgtttgggg aaagaccaca
     5401 cccacgcgcg atcgccatgg caacgaggtc gcacacgccc cacacccaga cctccctgcg
     5461 agcgggcatg ggtacaatgt ccccgttgcc acagacacca cttcgtagca cagcgcagag
     5521 cgtagcgtgt tgttgctgct gacaaaagaa aatttttctt agcaaagcaa aggaggggaa
     5581 gcacgggcag atagcaccgt accataccct tggaaactcg aaatgaacga agcaggaaat
     5641 gagagaatga gagttttgta ggtatatata gcggtagtgt ttgcgcgtta ccatcatctt
     5701 ctggatctat ctattgttct tttcctcatc actttcccct ttttcgctct tcttcttgtc
     5761 ttttatttct ttcttttttt taattgttcc ctcgattggc tatctaccaa agaatccaaa
     5821 cttaatacac gtatttattt gtccaattac catggataag aaatactcaa taggcttaga
     5881 tatcggcaca aatagcgtcg gatgggcggt gatcactgat gaatataagg ttccgtctaa
     5941 aaagttcaag gttctgggaa atacagaccg ccacagtatc aaaaaaaatc ttataggggc
     6001 tcttttattt gacagtggag agacagcgga agcgactcga ctcaaacgga cagctcgtag
     6061 aaggtataca cgtcggaaga atcgtatttg ttatctacag gagatttttt caaatgagat
     6121 ggcgaaagta gatgatagtt tctttcatcg acttgaagag tcttttttgg tggaagaaga
     6181 caagaagcat gaacgtcatc ctatttttgg aaatatagta gatgaagttg cttatcatga
     6241 gaaatatcca actatctatc atctgcgaaa aaaattggta gattctactg ataaagcgga
     6301 tttgcgctta atctatttgg ccttagcgca tatgattaag tttcgtggtc attttttgat
     6361 tgagggagat ttaaatcctg ataatagtga tgtggacaaa ctatttatcc agttggtaca
     6421 aacctacaat caattatttg aagaaaaccc tattaacgca agtggagtag atgctaaagc
     6481 gattctttct gcacgattga gtaaatcaag acgattagaa aatctcattg ctcagctccc
     6541 cggtgagaag aaaaatggct tatttgggaa tctcattgct ttgtcattgg gtttgacccc
     6601 taattttaaa tcaaattttg atttggcaga agatgctaaa ttacagcttt caaaagatac
     6661 ttacgatgat gatttagata atttattggc gcaaattgga gatcaatatg ctgatttgtt
     6721 tttggcagct aagaatttat cagatgctat tttactttca gatatcctaa gagtaaatac
     6781 tgaaataact aaggctcccc tatcagcttc aatgattaaa cgctacgatg aacatcatca
     6841 agacttgact cttttaaaag ctttagttcg acaacaactt ccagaaaagt ataaagaaat
     6901 cttttttgat caatcaaaaa acggatatgc aggttatatt gatgggggag ctagccaaga
     6961 agaattttat aaatttatca aaccaatttt agaaaaaatg gatggtactg aggaattatt
     7021 ggtgaaacta aatcgtgaag atttgctgcg caagcaacgg acctttgaca acggctctat
     7081 tccccatcaa attcacttgg gtgagctgca tgctattttg agaagacaag aagactttta
     7141 tccattttta aaagacaatc gtgagaagat tgaaaaaatc ttgacttttc gaattcctta
     7201 ttatgttggt ccattggcgc gtggcaatag tcgttttgca tggatgactc ggaagtctga
     7261 agaaacaatt accccatgga attttgaaga agttgtcgat aaaggtgctt cagctcaatc
     7321 atttattgaa cgcatgacaa actttgataa aaatcttcca aatgaaaaag tactaccaaa
     7381 acatagtttg ctttatgagt attttacggt ttataacgaa ttgacaaagg tcaaatatgt
     7441 tactgaagga atgcgaaaac cagcatttct ttcaggtgaa cagaagaaag ccattgttga
     7501 tttactcttc aaaacaaatc gaaaagtaac cgttaagcaa ttaaaagaag attatttcaa
     7561 aaaaatagaa tgttttgata gtgttgaaat ttcaggagtt gaagatagat ttaatgcttc
     7621 attaggtacc taccatgatt tgctaaaaat tattaaagat aaagattttt tggataatga
     7681 agaaaatgaa gatatcttag aggatattgt tttaacattg accttatttg aagataggga
     7741 gatgattgag gaaagactta aaacatatgc tcacctcttt gatgataagg tgatgaaaca
     7801 gcttaaacgt cgccgttata ctggttgggg acgtttgtct cgaaaattga ttaatggtat
     7861 tagggataag caatctggca aaacaatatt agattttttg aaatcagatg gttttgccaa
     7921 tcgcaatttt atgcagctga tccatgatga tagtttgaca tttaaagaag acattcaaaa
     7981 agcacaagtg tctggacaag gcgatagttt acatgaacat attgcaaatt tagctggtag
     8041 ccctgctatt aaaaaaggta ttttacagac tgtaaaagtt gttgatgaat tggtcaaagt
     8101 aatggggcgg cataagccag aaaatatcgt tattgaaatg gcacgtgaaa atcagacaac
     8161 tcaaaagggc cagaaaaatt cgcgagagcg tatgaaacga atcgaagaag gtatcaaaga
     8221 attaggaagt cagattctta aagagcatcc tgttgaaaat actcaattgc aaaatgaaaa
     8281 gctctatctc tattatctcc aaaatggaag agacatgtat gtggaccaag aattagatat
     8341 taatcgttta agtgattatg atgtcgatca cattgttcca caaagtttcc ttaaagacga
     8401 ttcaatagac aataaggtct taacgcgttc tgataaaaat cgtggtaaat cggataacgt
     8461 tccaagtgaa gaagtagtca aaaagatgaa aaactattgg agacaacttc taaacgccaa
     8521 gttaatcact caacgtaagt ttgataattt aacgaaagct gaacgtggag gtttgagtga
     8581 acttgataaa gctggtttta tcaaacgcca attggttgaa actcgccaaa tcactaagca
     8641 tgtggcacaa attttggata gtcgcatgaa tactaaatac gatgaaaatg ataaacttat
     8701 tcgagaggtt aaagtgatta ccttaaaatc taaattagtt tctgacttcc gaaaagattt
     8761 ccaattctat aaagtacgtg agattaacaa ttaccatcat gcccatgatg cgtatctaaa
     8821 tgccgtcgtt ggaactgctt tgattaagaa atatccaaaa cttgaatcgg agtttgtcta
     8881 tggtgattat aaagtttatg atgttcgtaa aatgattgct aagtctgagc aagaaatagg
     8941 caaagcaacc gcaaaatatt tcttttactc taatatcatg aacttcttca aaacagaaat
     9001 tacacttgca aatggagaga ttcgcaaacg ccctctaatc gaaactaatg gggaaactgg
     9061 agaaattgtc tgggataaag ggcgagattt tgccacagtg cgcaaagtat tgtccatgcc
     9121 ccaagtcaat attgtcaaga aaacagaagt acagacaggc ggattctcca aggagtcaat
     9181 tttaccaaaa agaaattcgg acaagcttat tgctcgtaaa aaagactggg atccaaaaaa
     9241 atatggtggt tttgatagtc caacggtagc ttattcagtc ctagtggttg ctaaggtgga
     9301 aaaagggaaa tcgaagaagt taaaatccgt taaagagtta ctagggatca caattatgga
     9361 aagaagttcc tttgaaaaaa atccgattga ctttttagaa gctaaaggat ataaggaagt
     9421 taaaaaagac ttaatcatta aactacctaa atatagtctt tttgagttag aaaacggtcg
     9481 taaacggatg ctggctagtg ccggagaatt acaaaaagga aatgagctgg ctctgccaag
     9541 caaatatgtg aattttttat atttagctag tcattatgaa aagttgaagg gtagtccaga
     9601 agataacgaa caaaaacaat tgtttgtgga gcagcataag cattatttag atgagattat
     9661 tgagcaaatc agtgaatttt ctaagcgtgt tattttagca gatgccaatt tagataaagt
     9721 tcttagtgca tataacaaac atagagacaa accaatacgt gaacaagcag aaaatattat
     9781 tcatttattt acgttgacga atcttggagc tcccgctgct tttaaatatt ttgatacaac
     9841 aattgatcgt aaacgatata cgtctacaaa agaagtttta gatgccactc ttatccatca
     9901 atccatcact ggtctttatg aaacacgcat tgatttgagt cagctaggag gtgacggtgg
     9961 agggccaaaa aagaaaagaa aagttgaaga tgcttctgct catcatcacc atcaccatca
    10021 ccattaaatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta
    10081 tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
    10141 tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg
    10201 ttttgggacg ctcgaagtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
    10261 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
    10321 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
    10381 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
    10441 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
    10501 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
    10561 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
    10621 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
    10681 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
    10741 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
    10801 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
    10861 aagaagatcc tttgatcttt tctac
//
