LOCUS       sequence-193261-       10958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GAL1 driven Cas9. gRNA can be cloned into BplI sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10958)
  AUTHORS   Anand R, Beach A, Li K, Haber J
  TITLE     Rad51-mediated double-strand break repair and mismatch correction of
            divergent substrates.
  JOURNAL   Nature. 2017 Apr 20;544(7650):377-380. doi: 10.1038/nature22046. 
            Epub 2017 Apr 12.
  PUBMED    28405019
REFERENCE   2  (bases 1 to 10958)
  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"; doi: 
            "10.1038/nature22046"; journalName: "Nature"; date: "2017-04-20- 
            20"; volume: "544"; issue: "7650"; pages: "377-380"
COMMENT     Sequence Label: bRA66
FEATURES             Location/Qualifiers
     source          1..10958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     gene            complement(284..1859)
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     terminator      complement(284..481)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(487..1515)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             complement(487..1515)
                     /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"
     promoter        complement(1516..1859)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        1993..2434
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1993..2110
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2405..2428
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2454..6557
                     /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             2454..6557
                     /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             6570..6590
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6594..6614
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6618..6638
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6639..6704
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      6723..6970
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6739..6757)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(7030..7485)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7167..7188)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7379..7398
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7611..7633
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7625..7642
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7626..7642
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7649..7668
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7649..7667
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7700..7716
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7700..7716
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        7748..8016
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        8044..8118
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      8123..8142
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        complement(8185..8203)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8224..8240)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8224..8240)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8237..8259)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8248..8264
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8272..8278,8279..8296,8297..8302))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8317..8338
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8455..8472)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8626..9214)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8706..8725)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9385..10176,10177..10245))
                     /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     10008..10027
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10246..10350)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    10387..10890
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     10931..10949
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcgaattcg
      241 agctcgttta aactggatgg cggcgttagt atcgaatcga cagcagtata gcgaccagca
      301 ttcacatacg attgacgcat gatattactt tctgcgcact taacttcgca tctgggcaga
      361 tgatgtcgag gcgaaaaaaa atataaatca cgctaacatt tgattaaaat agaacaacta
      421 caatataaaa aaactataca aatgacaagt tcttgaaaac aagaatcttt ttattgtcag
      481 tactgattat tcctttgccc tcggacgagt gctggggcgt cggtttccac tatcggcgag
      541 tacttctaca cagccatcgg tccagacggc cgcgcttctg cgggcgattt gtgtacgccc
      601 gacagtcccg gctccggatc ggacgattgc gtcgcatcga ccctgcgccc aagctgcatc
      661 atcgaaattg ccgtcaacca agctctgata gagttggtca agaccaatgc ggagcatata
      721 cgcccggagc cgcggcgatc ctgcaagctc cggatgcctc cgctcgaagt agcgcgtctg
      781 ctgctccata caagccaacc acggcctcca gaagaagatg ttggcgacct cgtattggga
      841 atccccgaac atcgcctcgc tccagtcaat gaccgctgtt atgcggccat tgtccgtcag
      901 gacattgttg gagccgaaat ccgcgtgcac gaggtgccgg acttcggggc agtcctcggc
      961 ccaaagcatc agctcatcga gagcctgcgc gacggacgca ctgacggtgt cgtccatcac
     1021 agtttgccag tgatacacat ggggatcagc aatcgcgcat atgaaatcac gccatgtagt
     1081 gtattgaccg attccttgcg gtccgaatgg gccgaacccg ctcgtctggc taagatcggc
     1141 cgcagcgatc gcatccatgg cctccgcgac cggctgcaga acagcgggca gttcggtttc
     1201 aggcaggtct tgcaacgtga caccctgtgc acggcgggag atgcaatagg tcaggctctc
     1261 gctgaattcc ccaatgtcaa gcacttccgg aatcgggagc gcggccgatg caaagtgccg
     1321 ataaacataa cgatctttgt agaaaccatc ggcgcagcta tttacccgca ggacatatcc
     1381 acgccctcct acatcgaagc tgaaagcacg agattcttcg ccctccgaga gctgcatcag
     1441 gtcggagacg ctgtcgaact tttcgatcag aaacttctcg acagacgtcg cggtgagttc
     1501 aggcttttta cccatggttg tttatgttcg gatgtgatgt gagaactgta tcctagcaag
     1561 attttaaaag gaagtatatg aaagaagaac ctcagtggca aatcctaacc ttttatattt
     1621 ctctacaggg gcgcggcgtg gggacaattc aacgcgtctg tgaggggagc gtttccctgc
     1681 tcgcaggtcc gcagcgagga gccgtaattt ttgcttcgcg ccgtgcggcc atcaaaatgt
     1741 atggatgcaa atgattatac atggggatgt atgggctaaa tgtacgggcg acagtcacat
     1801 catgcccctg agctgcgcac gtcaagactg tcaaggaggg tattctgggc ctccatgtcg
     1861 ctggccgggt gacccggcgg ggacgaggca agctaaacag atctgtaaag agccccatta
     1921 tcttagccta aaaaaacctt ctctttggaa ctttcagtaa tacgcttaac tgctcattgc
     1981 tatattgaag tacggattag aagccgccga gcgggtgaca gccctccgaa ggaagactct
     2041 cctccgtgcg tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg cctcgcgccg
     2101 cactgctccg aacaataaag attctacaat actagctttt atggttatga agaggaaaaa
     2161 ttggcagtaa cctggcccca caaaccttca aatgaacgaa tcaaattaac aaccatagga
     2221 tgataatgcg attagttttt tagccttatt tctggggtaa ttaatcagcg aagcgatgat
     2281 ttttgatcta ttaacagata tataaatgca aaaactgcat aaccacttta actaatactt
     2341 tcaacatttt cggtttgtat tacttcttat tcaaatgtaa taaaagtatc aacaaaaaat
     2401 tgttaatata cctctatact ttaacgtcaa ggagaaaaaa cccggatctc aaaatggata
     2461 aaaagtatag tattggttta gatattggta ctaactctgt gggttgggca gttatcactg
     2521 acgaatataa agttccatca aagaaattta aggtgttagg taacactgac agacactcaa
     2581 taaaaaagaa tcttatcggt gctcttttgt tcgactccgg tgaaactgcc gaggctacac
     2641 gtttaaaaag aacagcaaga agaagatata cccgtagaaa aaatagaata tgttatttac
     2701 aagaaatctt ttctaatgaa atggctaaag ttgatgattc ctttttccat agattggaag
     2761 agtcattttt ggttgaagaa gacaaaaagc atgagagaca tccaatcttt gggaatatag
     2821 ttgatgaagt ggcttaccat gaaaaatatc ctaccattta tcatttaaga aagaaattgg
     2881 tagattcaac tgataaagct gaccttagat taatctattt agcacttgcc catatgatta
     2941 aatttagagg tcattttttg attgaaggtg atttgaaccc agataattct gacgtggata
     3001 aattatttat tcaattagtc caaacctaca accaattatt tgaggaaaat ccaattaatg
     3061 ctagtggtgt cgatgccaaa gctatattat cagccagatt atcaaaatct agacgtttgg
     3121 aaaatttgat tgcccaattg ccaggagaaa aaaagaatgg attatttgga aacttgatcg
     3181 cattatcatt gggtttgaca ccaaatttta aatctaattt tgatttagct gaagatgcta
     3241 aattacaatt atcaaaagac acctatgacg acgatttgga caatttactt gctcaaattg
     3301 gtgatcaata tgcagatttg ttcttagctg ctaaaaactt atctgatgct attttgttgt
     3361 ctgatatttt gagagtgaac acagaaataa ccaaagctcc attatcagca tctatgatca
     3421 aacgttatga tgaacaccat caggatttga ctttattgaa agctttggtg agacaacaat
     3481 tgccagagaa gtataaagaa atctttttcg atcaatctaa aaacgggtat gcaggttata
     3541 ttgatggggg tgcctcccaa gaggaatttt acaaatttat aaaacctatt ttagaaaaga
     3601 tggatgggac tgaggaactt ttggtcaaat tgaacagaga agatttgtta cgtaaacaga
     3661 gaacttttga taatggtagt atacctcacc aaattcattt gggtgagttg catgcaattt
     3721 taagaagaca agaagatttt tatccatttt taaaagataa tagagaaaaa atcgagaaaa
     3781 ttttaacctt tagaattcca tactatgttg ggcctttggc tagaggtaat tcaagatttg
     3841 cctggatgac acgtaaatca gaagaaacta ttaccccttg gaattttgaa gaggttgttg
     3901 ataaaggagc atcagcacag agttttattg aaagaatgac caatttcgat aaaaacttac
     3961 caaatgaaaa agttttacca aaacattcct tgttatacga atattttact gtttacaatg
     4021 aacttacaaa ggttaaatat gttactgaag gtatgcgtaa gccagccttt ttatctggag
     4081 aacagaaaaa ggcaatagtt gatttattgt ttaaaacaaa tagaaaagtt actgttaaac
     4141 aattaaaaga agattacttt aagaaaattg aatgttttga ttcagttgaa atcagtggtg
     4201 ttgaagacag atttaatgct agtttaggaa cttaccatga tttacttaaa attatcaaag
     4261 ataaagattt cttggataac gaagaaaatg aagacatttt agaagacatt gttttaacct
     4321 taactttatt cgaagataga gagatgattg aagaacgttt gaagacttat gcacatttgt
     4381 ttgacgataa agtgatgaaa cagttgaaaa gaagacgtta tactggatgg ggtagattgt
     4441 ctcgtaaatt gatcaatgga attagagata aacaaagtgg taaaactatc ttggactttt
     4501 tgaaatctga cggatttgct aatagaaatt tcatgcaatt gatccacgac gatagtttga
     4561 catttaaaga agacatccaa aaggcccaag tgagtgggca aggtgattca ttacatgaac
     4621 atattgcaaa tttagccgga tctcctgcta ttaagaaagg gatattacaa actgttaaag
     4681 ttgtggatga attagtgaaa gtaatgggaa gacataaacc tgaaaacatt gtcattgaga
     4741 tggcaagaga aaatcaaact acacaaaaag gacagaaaaa tagtagagaa cgtatgaaaa
     4801 gaatagaaga gggtattaaa gaattgggta gtcaaatatt gaaagaacac ccagtggaaa
     4861 atacccagtt gcaaaatgaa aaattatatc tttactacct tcaaaatgga cgtgatatgt
     4921 atgttgatca ggaattagat ataaatagac tttcagatta tgatgtagat catatagttc
     4981 cacaatcttt cttgaaagat gattccatag acaataaagt attaactaga agtgataaaa
     5041 atagaggtaa aagtgataat gtcccaagtg aggaagtcgt caaaaagatg aaaaattact
     5101 ggcgtcaact tttgaatgct aaattaatta ctcaaagaaa atttgataat ttgactaaag
     5161 cagaaagagg tgggctttct gaattagata aagccgggtt cattaaaaga caattggtcg
     5221 aaactagaca aattactaaa catgttgccc aaattttaga ttcccgtatg aacactaagt
     5281 atgacgaaaa tgataagtta atacgtgagg ttaaagtcat tactttaaaa tcaaaacttg
     5341 tctctgattt cagaaaggat ttccaattct ataaagttag agaaattaat aattatcatc
     5401 atgctcatga tgcatatttg aatgctgtag ttggaactgc tttaatcaag aaatacccta
     5461 aattagaatc tgaatttgta tatggtgatt acaaagtcta tgatgttaga aagatgattg
     5521 ctaaatcaga acaagaaatt ggtaaagcta cagctaaata cttcttttac tctaacatta
     5581 tgaatttctt taaaacagaa attactttgg caaacggtga aattagaaaa agacctctta
     5641 ttgaaacaaa tggtgagact ggagagatag tttgggacaa agggcgtgat ttcgctactg
     5701 ttagaaaagt tttatcaatg ccacaagtta acattgtaaa gaaaacagag gttcaaactg
     5761 gtggtttctc aaaagaaagt attttgccta aaagaaatag tgataaattg attgccagaa
     5821 aaaaggattg ggatccaaag aaatatggtg gtttcgactc accaaccgta gcctattctg
     5881 ttttggttgt ggcaaaggtt gaaaagggta aaagtaaaaa gcttaaatca gtaaaagaac
     5941 ttttgggtat tacaataatg gaaagaagtt cctttgaaaa gaaccctatt gattttttgg
     6001 aagctaaagg ttataaggaa gtaaagaagg acttaataat caaattgcct aaatattctt
     6061 tatttgaatt agaaaatggg agaaaaagaa tgttggcttc tgctggagaa ttgcaaaagg
     6121 gtaatgaatt agcattgcct tccaaatatg ttaacttctt gtatttagct tcacactatg
     6181 aaaagttgaa agggtcacca gaagataacg agcaaaaaca attatttgtt gaacaacaca
     6241 aacactactt agatgagatt atagaacaaa ttagtgaatt cagtaaaaga gtgatattag
     6301 ctgatgcaaa tttagataaa gttttgtcag cctataacaa acatagagat aagccaatta
     6361 gagaacaagc agaaaacatt attcacttat ttacccttac caatttagga gcacctgctg
     6421 ctttcaagta ttttgataca acaattgatc gtaaaagata tacctcaaca aaagaagtct
     6481 tagacgccac cttaattcat caatcaatca ctggattgta tgagacaaga attgatttgt
     6541 ctcaattggg tggtgatgaa ggggctgatc ctaagaagaa aagaaaagtt gatccaaaga
     6601 aaaagcgtaa ggtggatcct aagaaaaaga gaaaggttga ctacaaagac catgacggtg
     6661 attataaaga tcatgacatc gactacaagg atgacgatga caagtgataa tctcttctcg
     6721 agtcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac
     6781 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat
     6841 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt
     6901 gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc
     6961 tttaatttgc ggccggtact gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     7021 cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     7081 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     7141 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     7201 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     7261 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     7321 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     7381 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     7441 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag
     7501 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     7561 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     7621 acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg aattggagct
     7681 ccaccgcggt ggcggccgct ctagaactag tggatccccc gggctgcagg aattcgatat
     7741 caagctttct ttgaaaagat aatgtatgat tatgctttca ctcatattta tacagaaact
     7801 tgatgttttc tttcgagtat atacaaggtg attacatgta cgtttgaagt acaactctag
     7861 attttgtagt gccctcttgg gctagcggta aaggtgcgca ttttttcaca ccctacaatg
     7921 ttctgttcaa aagattttgg tcaaacgctg tagaagtgaa agttggtgcg catgtttcgg
     7981 cgttcgaaac ttctccgcag tgaaagataa atgatcctag atgcgagata tcctcatgcc
     8041 taggttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
     8101 agtggcaccg agtcggtggt gctttttttg ttttttatgt ctaccgtcga cctcgagggg
     8161 gggcccggta cccagctttt gttcccttta gtgagggtta atttcgagct tggcgtaatc
     8221 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     8281 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     8341 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     8401 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     8461 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     8521 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     8581 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     8641 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     8701 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     8761 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     8821 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     8881 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     8941 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     9001 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     9061 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     9121 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     9181 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     9241 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     9301 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     9361 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     9421 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     9481 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     9541 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     9601 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     9661 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     9721 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     9781 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     9841 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     9901 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     9961 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
    10021 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
    10081 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
    10141 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
    10201 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
    10261 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
    10321 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggtc
    10381 cttttcatca cgtgctataa aaataattat aatttaaatt ttttaatata aatatataaa
    10441 ttaaaaatag aaagtaaaaa aagaaattaa agaaaaaata gtttttgttt tccgaagatg
    10501 taaaagactc tagggggatc gccaacaaat actacctttt atcttgctct tcctgctctc
    10561 aggtattaat gccgaattgt ttcatcttgt ctgtgtagaa gaccacacac gaaaatcctg
    10621 tgattttaca ttttacttat cgttaatcga atgtatatct atttaatctg cttttcttgt
    10681 ctaataaata tatatgtaaa gtacgctttt tgttgaaatt ttttaaacct ttgtttattt
    10741 ttttttcttc attccgtaac tcttctacct tctttattta ctttctaaaa tccaaataca
    10801 aaacataaaa ataaataaac acagagtaaa ttcccaaatt attccatcat taaaagatac
    10861 gaggcgcgtg taagttacag gcaagcgatc cgtctaagaa accattatta tcatgacatt
    10921 aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
