LOCUS       sequence-368820-        8982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sp.Cas9 and gRNA yeast expression vector for HDR-based gRNA 
            introduction. Selection = Hygromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8982)
  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 8982)
  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: pBBK95 pCAS-Tyr-[gRNA: 2xBsaI+NotI cut sites] ...
FEATURES             Location/Qualifiers
     source          1..8982
                     /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)."
     misc_RNA        1402..1477
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     gene            1611..3186
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        1611..1954
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1955..2983
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             1955..2983
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2989..3186
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             3949..8052
                     /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             3949..8052
                     /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             8062..8082
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8098..8121
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      8125..8314
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(8371..8959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8451..8470)
                     /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 aatgggactt tcgagaccaa ggtattgttt
     1381 gcggccgctt tacaggtctc cgttttagag ctagaaatag caagttaaaa taaggctagt
     1441 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttattttt tgtcactatt
     1501 gttatgtaaa atgccacctc tgacagtatg gaacgcaaac ttctgtctag tggatatagg
     1561 tctagagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg
     1621 cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc
     1681 gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata cattttgatg
     1741 gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg
     1801 ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag
     1861 gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat
     1921 acagttctca catcacatcc gaacataaac aaccatgggt aaaaagcctg aactcaccgc
     1981 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
     2041 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
     2101 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     2161 atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac
     2221 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
     2281 gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag
     2341 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     2401 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     2461 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     2521 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     2581 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     2641 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     2701 cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg
     2761 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     2821 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     2881 aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
     2941 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa taatcagtac tgacaataaa
     3001 aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta
     3061 ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat
     3121 gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct
     3181 atactgctgt cgattcgata ctaacgccgc catccagtgt cgaaaacgag ctctcgagaa
     3241 cccttaatag tcgaacaaga agcaggcaaa gtttagagca ctgcccctcc gcactcaaaa
     3301 aagaaaaaac taggaggaaa ataaaattct caaccacaca aacacataaa cacatacaaa
     3361 tacaaataca agcttattta cttgacatcg cgcgatcttc cactattcag cgccgtccgc
     3421 cctctctcgt gttttttgtt tacgcgacaa ctatgcgaaa tccggagcaa cgggcaaccg
     3481 tttggggaaa gaccacaccc acgcgcgatc gccatggcaa cgaggtcgca cacgccccac
     3541 acccagacct ccctgcgagc gggcatgggt acaatgtccc cgttgccaca gacaccactt
     3601 cgtagcacag cgcagagcgt agcgtgttgt tgctgctgac aaaagaaaat ttttcttagc
     3661 aaagcaaagg aggggaagca cgggcagata gcaccgtacc atacccttgg aaactcgaaa
     3721 tgaacgaagc aggaaatgag agaatgagag ttttgtaggt atatatagcg gtagtgtttg
     3781 cgcgttacca tcatcttctg gatctatcta ttgttctttt cctcatcact ttcccctttt
     3841 tcgctcttct tcttgtcttt tatttctttc ttttttttaa ttgttccctc gattggctat
     3901 ctaccaaaga atccaaactt aatacacgta tttatttgtc caattaccat ggataagaaa
     3961 tactcaatag gcttagatat cggcacaaat agcgtcggat gggcggtgat cactgatgaa
     4021 tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca cagtatcaaa
     4081 aaaaatctta taggggctct tttatttgac agtggagaga cagcggaagc gactcgtctc
     4141 aaacggacag ctcgtagaag gtatacacgt cggaagaatc gtatttgtta tctacaggag
     4201 attttttcaa atgagatggc gaaagtagat gatagtttct ttcatcgact tgaagagtct
     4261 tttttggtgg aagaagacaa gaagcatgaa cgtcatccta tttttggaaa tatagtagat
     4321 gaagttgctt atcatgagaa atatccaact atctatcatc tgcgaaaaaa attggtagat
     4381 tctactgata aagcggattt gcgcttaatc tatttggcct tagcgcatat gattaagttt
     4441 cgtggtcatt ttttgattga gggagattta aatcctgata atagtgatgt ggacaaacta
     4501 tttatccagt tggtacaaac ctacaatcaa ttatttgaag aaaaccctat taacgcaagt
     4561 ggagtagatg ctaaagcgat tctttctgca cgattgagta aatcaagacg attagaaaat
     4621 ctcattgctc agctccccgg tgagaagaaa aatggcttat ttgggaatct cattgctttg
     4681 tcattgggtt tgacccctaa ttttaaatca aattttgatt tggcagaaga tgctaaatta
     4741 cagctttcaa aagatactta cgatgatgat ttagataatt tattggcgca aattggagat
     4801 caatatgctg atttgttttt ggcagctaag aatttatcag atgctatttt actttcagat
     4861 atcctaagag taaatactga aataactaag gctcccctat cagcttcaat gattaaacgc
     4921 tacgatgaac atcatcaaga cttgactctt ttaaaagctt tagttcgaca acaacttcca
     4981 gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg ttatattgat
     5041 gggggagcta gccaagaaga attttataaa tttatcaaac caattttaga aaaaatggat
     5101 ggtactgagg aattattggt gaaactaaat cgtgaagatt tgctgcgcaa gcaacggacc
     5161 tttgacaacg gctctattcc ccatcaaatt cacttgggtg agctgcatgc tattttgaga
     5221 agacaagaag acttttatcc atttttaaaa gacaatcgtg agaagattga aaaaatcttg
     5281 acttttcgaa ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg ttttgcatgg
     5341 atgactcgga agtctgaaga aacaattacc ccatggaatt ttgaagaagt tgtcgataaa
     5401 ggtgcttcag ctcaatcatt tattgaacgc atgacaaact ttgataaaaa tcttccaaat
     5461 gaaaaagtac taccaaaaca tagtttgctt tatgagtatt ttacggttta taacgaattg
     5521 acaaaggtca aatatgttac tgaaggaatg cgaaaaccag catttctttc aggtgaacag
     5581 aagaaagcca ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt taagcaatta
     5641 aaagaagatt atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc aggagttgaa
     5701 gatagattta atgcttcatt aggtacctac catgatttgc taaaaattat taaagataaa
     5761 gattttttgg ataatgaaga aaatgaagat atcttagagg atattgtttt aacattgacc
     5821 ttatttgaag atagggagat gattgaggaa agacttaaaa catatgctca cctctttgat
     5881 gataaggtga tgaaacagct taaacgtcgc cgttatactg gttggggacg tttgtctcga
     5941 aaattgatta atggtattag ggataagcaa tctggcaaaa caatattaga ttttttgaaa
     6001 tcagatggtt ttgccaatcg caattttatg cagctgatcc atgatgatag tttgacattt
     6061 aaagaagaca ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca tgaacatatt
     6121 gcaaatttag ctggtagccc tgctattaaa aaaggtattt tacagactgt aaaagttgtt
     6181 gatgaattgg tcaaagtaat ggggcggcat aagccagaaa atatcgttat tgaaatggca
     6241 cgtgaaaatc agacaactca aaagggccag aaaaattcgc gagagcgtat gaaacgaatc
     6301 gaagaaggta tcaaagaatt aggaagtcag attcttaaag agcatcctgt tgaaaatact
     6361 caattgcaaa atgaaaagct ctatctctat tatctccaaa atggaagaga catgtatgtg
     6421 gaccaagaat tagatattaa tcgtttaagt gattatgatg tcgatcacat tgttccacaa
     6481 agtttcctta aagacgattc aatagacaat aaggtcttaa cgcgttctga taaaaatcgt
     6541 ggtaaatcgg ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa ctattggaga
     6601 caacttctaa acgccaagtt aatcactcaa cgtaagtttg ataatttaac gaaagctgaa
     6661 cgtggaggtt tgagtgaact tgataaagct ggttttatca aacgccaatt ggttgaaact
     6721 cgccaaatca ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac taaatacgat
     6781 gaaaatgata aacttattcg agaggttaaa gtgattacct taaaatctaa attagtttct
     6841 gacttccgaa aagatttcca attctataaa gtacgtgaga ttaacaatta ccatcatgcc
     6901 catgatgcgt atctaaatgc cgtcgttgga actgctttga ttaagaaata tccaaaactt
     6961 gaatcggagt ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat gattgctaag
     7021 tctgagcaag aaataggcaa agcaaccgca aaatatttct tttactctaa tatcatgaac
     7081 ttcttcaaaa cagaaattac acttgcaaat ggagagattc gcaaacgccc tctaatcgaa
     7141 actaatgggg aaactggaga aattgtctgg gataaagggc gagattttgc cacagtgcgc
     7201 aaagtattgt ccatgcccca agtcaatatt gtcaagaaaa cagaagtaca gacaggcgga
     7261 ttctccaagg agtcaatttt accaaaaaga aattcggaca agcttattgc tcgtaaaaaa
     7321 gactgggatc caaaaaaata tggtggtttt gatagtccaa cggtagctta ttcagtccta
     7381 gtggttgcta aggtggaaaa agggaaatcg aagaagttaa aatccgttaa agagttacta
     7441 gggatcacaa ttatggaaag aagttccttt gaaaaaaatc cgattgactt tttagaagct
     7501 aaaggatata aggaagttaa aaaagactta atcattaaac tacctaaata tagtcttttt
     7561 gagttagaaa acggtcgtaa acggatgctg gctagtgccg gagaattaca aaaaggaaat
     7621 gagctggctc tgccaagcaa atatgtgaat tttttatatt tagctagtca ttatgaaaag
     7681 ttgaagggta gtccagaaga taacgaacaa aaacaattgt ttgtggagca gcataagcat
     7741 tatttagatg agattattga gcaaatcagt gaattttcta agcgtgttat tttagcagat
     7801 gccaatttag ataaagttct tagtgcatat aacaaacata gagacaaacc aatacgtgaa
     7861 caagcagaaa atattattca tttatttacg ttgacgaatc ttggagctcc cgctgctttt
     7921 aaatattttg atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga agttttagat
     7981 gccactctta tccatcaatc catcactggt ctttatgaaa cacgcattga tttgagtcag
     8041 ctaggaggtg acggtggagg gccaaaaaag aaaagaaaag ttgaagatgc ttctgctcat
     8101 catcaccatc accatcacca ttaaatccgc tctaaccgaa aaggaaggag ttagacaacc
     8161 tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat
     8221 atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa
     8281 aaccttgctt gagaaggttt tgggacgctc gaagtgagca aaaggccagc aaaaggccag
     8341 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     8401 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     8461 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     8521 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     8581 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     8641 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     8701 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     8761 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     8821 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     8881 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     8941 cagaaaaaaa ggatctcaag aagatccttt gatcttttct ac
//
