LOCUS       sequence-360679-       11215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sp.Cas9 and gRNA yeast expression vector for Golden Gate pre-cloning
            of gRNA. Selection = SplitURA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11215)
  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 11215)
  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: pBBK10 pCAS-Tyr-[gRNA: BsaI GFP dropout]
            (Spli...
FEATURES             Location/Qualifiers
     source          1..11215
                     /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      4166..4363
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(join(4514..5305,5306..5374))
                     /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     5137..5156
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5375..5479)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6182..10285
                     /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             6182..10285
                     /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             10295..10315
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10331..10354
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      10358..10547
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(10604..11192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10684..10703)
                     /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 gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct
     3001 atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac
     3061 caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca
     3121 tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc
     3181 cgccaagtac aattttttac tcttcgagga cagaaaattt gctgacattg gtaatacagt
     3241 caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc
     3301 acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac
     3361 aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac
     3421 tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg
     3481 ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac
     3541 gtcgctgcct gcaggatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
     3601 ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt aatacagtca
     3661 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
     3721 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
     3781 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
     3841 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
     3901 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     3961 ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga accgtggatg
     4021 atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     4081 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     4141 gatgcggcca gcaaaactaa tcagtactga caataaaaag attcttgttt tcaagaactt
     4201 gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat
     4261 ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt
     4321 aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta
     4381 acgccgccat ccagtgtcga aaacgagctc tcgagaaccc ttaatcaaaa aggatcttca
     4441 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4501 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4561 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4621 taccatctgg ccccagtgct gcaatgatac cgcgggaccc acgctcaccg gctccagatt
     4681 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4741 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4801 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4861 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4921 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4981 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5041 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5101 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5161 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5221 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5281 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5341 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5401 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5461 aacaaatagg ggttcgtcga cagtcgaaca agaagcaggc aaagtttaga gcactgcccc
     5521 tccgcactca aaaaagaaaa aactaggagg aaaataaaat tctcaaccac acaaacacat
     5581 aaacacatac aaatacaaat acaagcttat ttacttgaca tcgcgcgatc ttccactatt
     5641 cagcgccgtc cgccctctct cgtgtttttt gtttacgcga caactatgcg aaatccggag
     5701 caacgggcaa ccgtttgggg aaagaccaca cccacgcgcg atcgccatgg caacgaggtc
     5761 gcacacgccc cacacccaga cctccctgcg agcgggcatg ggtacaatgt ccccgttgcc
     5821 acagacacca cttcgtagca cagcgcagag cgtagcgtgt tgttgctgct gacaaaagaa
     5881 aatttttctt agcaaagcaa aggaggggaa gcacgggcag atagcaccgt accataccct
     5941 tggaaactcg aaatgaacga agcaggaaat gagagaatga gagttttgta ggtatatata
     6001 gcggtagtgt ttgcgcgtta ccatcatctt ctggatctat ctattgttct tttcctcatc
     6061 actttcccct ttttcgctct tcttcttgtc ttttatttct ttcttttttt taattgttcc
     6121 ctcgattggc tatctaccaa agaatccaaa cttaatacac gtatttattt gtccaattac
     6181 catggataag aaatactcaa taggcttaga tatcggcaca aatagcgtcg gatgggcggt
     6241 gatcactgat gaatataagg ttccgtctaa aaagttcaag gttctgggaa atacagaccg
     6301 ccacagtatc aaaaaaaatc ttataggggc tcttttattt gacagtggag agacagcgga
     6361 agcgactcga ctcaaacgga cagctcgtag aaggtataca cgtcggaaga atcgtatttg
     6421 ttatctacag gagatttttt caaatgagat ggcgaaagta gatgatagtt tctttcatcg
     6481 acttgaagag tcttttttgg tggaagaaga caagaagcat gaacgtcatc ctatttttgg
     6541 aaatatagta gatgaagttg cttatcatga gaaatatcca actatctatc atctgcgaaa
     6601 aaaattggta gattctactg ataaagcgga tttgcgctta atctatttgg ccttagcgca
     6661 tatgattaag tttcgtggtc attttttgat tgagggagat ttaaatcctg ataatagtga
     6721 tgtggacaaa ctatttatcc agttggtaca aacctacaat caattatttg aagaaaaccc
     6781 tattaacgca agtggagtag atgctaaagc gattctttct gcacgattga gtaaatcaag
     6841 acgattagaa aatctcattg ctcagctccc cggtgagaag aaaaatggct tatttgggaa
     6901 tctcattgct ttgtcattgg gtttgacccc taattttaaa tcaaattttg atttggcaga
     6961 agatgctaaa ttacagcttt caaaagatac ttacgatgat gatttagata atttattggc
     7021 gcaaattgga gatcaatatg ctgatttgtt tttggcagct aagaatttat cagatgctat
     7081 tttactttca gatatcctaa gagtaaatac tgaaataact aaggctcccc tatcagcttc
     7141 aatgattaaa cgctacgatg aacatcatca agacttgact cttttaaaag ctttagttcg
     7201 acaacaactt ccagaaaagt ataaagaaat cttttttgat caatcaaaaa acggatatgc
     7261 aggttatatt gatgggggag ctagccaaga agaattttat aaatttatca aaccaatttt
     7321 agaaaaaatg gatggtactg aggaattatt ggtgaaacta aatcgtgaag atttgctgcg
     7381 caagcaacgg acctttgaca acggctctat tccccatcaa attcacttgg gtgagctgca
     7441 tgctattttg agaagacaag aagactttta tccattttta aaagacaatc gtgagaagat
     7501 tgaaaaaatc ttgacttttc gaattcctta ttatgttggt ccattggcgc gtggcaatag
     7561 tcgttttgca tggatgactc ggaagtctga agaaacaatt accccatgga attttgaaga
     7621 agttgtcgat aaaggtgctt cagctcaatc atttattgaa cgcatgacaa actttgataa
     7681 aaatcttcca aatgaaaaag tactaccaaa acatagtttg ctttatgagt attttacggt
     7741 ttataacgaa ttgacaaagg tcaaatatgt tactgaagga atgcgaaaac cagcatttct
     7801 ttcaggtgaa cagaagaaag ccattgttga tttactcttc aaaacaaatc gaaaagtaac
     7861 cgttaagcaa ttaaaagaag attatttcaa aaaaatagaa tgttttgata gtgttgaaat
     7921 ttcaggagtt gaagatagat ttaatgcttc attaggtacc taccatgatt tgctaaaaat
     7981 tattaaagat aaagattttt tggataatga agaaaatgaa gatatcttag aggatattgt
     8041 tttaacattg accttatttg aagataggga gatgattgag gaaagactta aaacatatgc
     8101 tcacctcttt gatgataagg tgatgaaaca gcttaaacgt cgccgttata ctggttgggg
     8161 acgtttgtct cgaaaattga ttaatggtat tagggataag caatctggca aaacaatatt
     8221 agattttttg aaatcagatg gttttgccaa tcgcaatttt atgcagctga tccatgatga
     8281 tagtttgaca tttaaagaag acattcaaaa agcacaagtg tctggacaag gcgatagttt
     8341 acatgaacat attgcaaatt tagctggtag ccctgctatt aaaaaaggta ttttacagac
     8401 tgtaaaagtt gttgatgaat tggtcaaagt aatggggcgg cataagccag aaaatatcgt
     8461 tattgaaatg gcacgtgaaa atcagacaac tcaaaagggc cagaaaaatt cgcgagagcg
     8521 tatgaaacga atcgaagaag gtatcaaaga attaggaagt cagattctta aagagcatcc
     8581 tgttgaaaat actcaattgc aaaatgaaaa gctctatctc tattatctcc aaaatggaag
     8641 agacatgtat gtggaccaag aattagatat taatcgttta agtgattatg atgtcgatca
     8701 cattgttcca caaagtttcc ttaaagacga ttcaatagac aataaggtct taacgcgttc
     8761 tgataaaaat cgtggtaaat cggataacgt tccaagtgaa gaagtagtca aaaagatgaa
     8821 aaactattgg agacaacttc taaacgccaa gttaatcact caacgtaagt ttgataattt
     8881 aacgaaagct gaacgtggag gtttgagtga acttgataaa gctggtttta tcaaacgcca
     8941 attggttgaa actcgccaaa tcactaagca tgtggcacaa attttggata gtcgcatgaa
     9001 tactaaatac gatgaaaatg ataaacttat tcgagaggtt aaagtgatta ccttaaaatc
     9061 taaattagtt tctgacttcc gaaaagattt ccaattctat aaagtacgtg agattaacaa
     9121 ttaccatcat gcccatgatg cgtatctaaa tgccgtcgtt ggaactgctt tgattaagaa
     9181 atatccaaaa cttgaatcgg agtttgtcta tggtgattat aaagtttatg atgttcgtaa
     9241 aatgattgct aagtctgagc aagaaatagg caaagcaacc gcaaaatatt tcttttactc
     9301 taatatcatg aacttcttca aaacagaaat tacacttgca aatggagaga ttcgcaaacg
     9361 ccctctaatc gaaactaatg gggaaactgg agaaattgtc tgggataaag ggcgagattt
     9421 tgccacagtg cgcaaagtat tgtccatgcc ccaagtcaat attgtcaaga aaacagaagt
     9481 acagacaggc ggattctcca aggagtcaat tttaccaaaa agaaattcgg acaagcttat
     9541 tgctcgtaaa aaagactggg atccaaaaaa atatggtggt tttgatagtc caacggtagc
     9601 ttattcagtc ctagtggttg ctaaggtgga aaaagggaaa tcgaagaagt taaaatccgt
     9661 taaagagtta ctagggatca caattatgga aagaagttcc tttgaaaaaa atccgattga
     9721 ctttttagaa gctaaaggat ataaggaagt taaaaaagac ttaatcatta aactacctaa
     9781 atatagtctt tttgagttag aaaacggtcg taaacggatg ctggctagtg ccggagaatt
     9841 acaaaaagga aatgagctgg ctctgccaag caaatatgtg aattttttat atttagctag
     9901 tcattatgaa aagttgaagg gtagtccaga agataacgaa caaaaacaat tgtttgtgga
     9961 gcagcataag cattatttag atgagattat tgagcaaatc agtgaatttt ctaagcgtgt
    10021 tattttagca gatgccaatt tagataaagt tcttagtgca tataacaaac atagagacaa
    10081 accaatacgt gaacaagcag aaaatattat tcatttattt acgttgacga atcttggagc
    10141 tcccgctgct tttaaatatt ttgatacaac aattgatcgt aaacgatata cgtctacaaa
    10201 agaagtttta gatgccactc ttatccatca atccatcact ggtctttatg aaacacgcat
    10261 tgatttgagt cagctaggag gtgacggtgg agggccaaaa aagaaaagaa aagttgaaga
    10321 tgcttctgct catcatcacc atcaccatca ccattaaatc cgctctaacc gaaaaggaag
    10381 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
    10441 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta
    10501 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaagtga gcaaaaggcc
    10561 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
    10621 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
    10681 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
    10741 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
    10801 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
    10861 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
    10921 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
    10981 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
    11041 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
    11101 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
    11161 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctac
//
