LOCUS       sequence-193243-       11380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 driven by PGK1 promoter. gRNA can be cloned into the BplI 
            sites. HPH marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11380)
  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 11380)
  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: bRA89
FEATURES             Location/Qualifiers
     source          1..11380
                     /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..1858)
                     /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..1858)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2876..6979
                     /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             2876..6979
                     /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             6992..7012
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7016..7036
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7040..7060
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7061..7126
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      7145..7392
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7161..7179)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(7452..7907)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7589..7610)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7801..7820
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8033..8055
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8047..8064
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8048..8064
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8071..8090
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8071..8089
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     8122..8138
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8122..8138
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        8170..8438
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        8466..8540
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      8545..8564
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        complement(8607..8625)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8646..8662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8646..8662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8659..8681)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8670..8686
                     /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(8694..8700,8701..8718,8719..8724))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8739..8760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8877..8894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9048..9636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9128..9147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9807..10598,10599..10667))
                     /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     10430..10449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10668..10772)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    10809..11312
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     11353..11371
                     /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 tcaaggaggt attctgggcc tccatgtcgc
     1861 tggccgggtg acccggcggg gaaagtacct tcaaagaatg gggtcttatc ttgttttgca
     1921 agtaccactg agcaggataa taatagaaat gataatatac tatagtagag ataacgtcga
     1981 tgacttccca tactgtaatt gcttttagtt gtgtattttt agtgtgcaag tttctgtaaa
     2041 tcgattaatt tttttttctt tcctcttttt attaacctta atttttattt tagattcctg
     2101 acttcaactc aagacgcaca gatattataa catctgcata ataggcattt gcaagaatta
     2161 ctcgtgagta aggaaagagt gaggaactat cgcatacctg catttaaaga tgccgatttg
     2221 ggcgcgaatc ctttattttg gcttcaccct catactatta tcagggccag aaaaaggaag
     2281 tgtttccctc cttcttgaat tgatgttacc ctcataaagc acgtggcctc ttatcgagaa
     2341 agaaattacc gtcgctcgtg atttgtttgc aaaaagaaca aaactgaaaa aacccagaca
     2401 cgctcgactt cctgtcttcc tattgattgc agcttccaat ttcgtcacac aacaaggtcc
     2461 tagcgacggc tcacaggttt tgtaacaagc aatcgaaggt tctggaatgg cgggaaaggg
     2521 tttagtacca catgctatga tgcccactgt gatctccaga gcaaagttcg ttcgatcgta
     2581 ctgttactct ctctctttca aacagaattg tccgaatcgt gtgacaacaa cagcctgttc
     2641 tcacacactc ttttcttcta accaaggggg tggtttagtt tagtagaacc tcgtgaaact
     2701 tacatttaca tatatataaa cttgcataaa ttggtcaatg caagaaatac atatttggtc
     2761 ttttctaatt cgtagttttt caagttctta gatgctttct ttttctcttt tttacagatc
     2821 atcaaggaag taattatcta ctttttacaa caaatataaa acaccggatc tcaaaatgga
     2881 taaaaagtat agtattggtt tagatattgg tactaactct gtgggttggg cagttatcac
     2941 tgacgaatat aaagttccat caaagaaatt taaggtgtta ggtaacactg acagacactc
     3001 aataaaaaag aatcttatcg gtgctctttt gttcgactcc ggtgaaactg ccgaggctac
     3061 acgtttaaaa agaacagcaa gaagaagata tacccgtaga aaaaatagaa tatgttattt
     3121 acaagaaatc ttttctaatg aaatggctaa agttgatgat tcctttttcc atagattgga
     3181 agagtcattt ttggttgaag aagacaaaaa gcatgagaga catccaatct ttgggaatat
     3241 agttgatgaa gtggcttacc atgaaaaata tcctaccatt tatcatttaa gaaagaaatt
     3301 ggtagattca actgataaag ctgaccttag attaatctat ttagcacttg cccatatgat
     3361 taaatttaga ggtcattttt tgattgaagg tgatttgaac ccagataatt ctgacgtgga
     3421 taaattattt attcaattag tccaaaccta caaccaatta tttgaggaaa atccaattaa
     3481 tgctagtggt gtcgatgcca aagctatatt atcagccaga ttatcaaaat ctagacgttt
     3541 ggaaaatttg attgcccaat tgccaggaga aaaaaagaat ggattatttg gaaacttgat
     3601 cgcattatca ttgggtttga caccaaattt taaatctaat tttgatttag ctgaagatgc
     3661 taaattacaa ttatcaaaag acacctatga cgacgatttg gacaatttac ttgctcaaat
     3721 tggtgatcaa tatgcagatt tgttcttagc tgctaaaaac ttatctgatg ctattttgtt
     3781 gtctgatatt ttgagagtga acacagaaat aaccaaagct ccattatcag catctatgat
     3841 caaacgttat gatgaacacc atcaggattt gactttattg aaagctttgg tgagacaaca
     3901 attgccagag aagtataaag aaatcttttt cgatcaatct aaaaacgggt atgcaggtta
     3961 tattgatggg ggtgcctccc aagaggaatt ttacaaattt ataaaaccta ttttagaaaa
     4021 gatggatggg actgaggaac ttttggtcaa attgaacaga gaagatttgt tacgtaaaca
     4081 gagaactttt gataatggta gtatacctca ccaaattcat ttgggtgagt tgcatgcaat
     4141 tttaagaaga caagaagatt tttatccatt tttaaaagat aatagagaaa aaatcgagaa
     4201 aattttaacc tttagaattc catactatgt tgggcctttg gctagaggta attcaagatt
     4261 tgcctggatg acacgtaaat cagaagaaac tattacccct tggaattttg aagaggttgt
     4321 tgataaagga gcatcagcac agagttttat tgaaagaatg accaatttcg ataaaaactt
     4381 accaaatgaa aaagttttac caaaacattc cttgttatac gaatatttta ctgtttacaa
     4441 tgaacttaca aaggttaaat atgttactga aggtatgcgt aagccagcct ttttatctgg
     4501 agaacagaaa aaggcaatag ttgatttatt gtttaaaaca aatagaaaag ttactgttaa
     4561 acaattaaaa gaagattact ttaagaaaat tgaatgtttt gattcagttg aaatcagtgg
     4621 tgttgaagac agatttaatg ctagtttagg aacttaccat gatttactta aaattatcaa
     4681 agataaagat ttcttggata acgaagaaaa tgaagacatt ttagaagaca ttgttttaac
     4741 cttaacttta ttcgaagata gagagatgat tgaagaacgt ttgaagactt atgcacattt
     4801 gtttgacgat aaagtgatga aacagttgaa aagaagacgt tatactggat ggggtagatt
     4861 gtctcgtaaa ttgatcaatg gaattagaga taaacaaagt ggtaaaacta tcttggactt
     4921 tttgaaatct gacggatttg ctaatagaaa tttcatgcaa ttgatccacg acgatagttt
     4981 gacatttaaa gaagacatcc aaaaggccca agtgagtggg caaggtgatt cattacatga
     5041 acatattgca aatttagccg gatctcctgc tattaagaaa gggatattac aaactgttaa
     5101 agttgtggat gaattagtga aagtaatggg aagacataaa cctgaaaaca ttgtcattga
     5161 gatggcaaga gaaaatcaaa ctacacaaaa aggacagaaa aatagtagag aacgtatgaa
     5221 aagaatagaa gagggtatta aagaattggg tagtcaaata ttgaaagaac acccagtgga
     5281 aaatacccag ttgcaaaatg aaaaattata tctttactac cttcaaaatg gacgtgatat
     5341 gtatgttgat caggaattag atataaatag actttcagat tatgatgtag atcatatagt
     5401 tccacaatct ttcttgaaag atgattccat agacaataaa gtattaacta gaagtgataa
     5461 aaatagaggt aaaagtgata atgtcccaag tgaggaagtc gtcaaaaaga tgaaaaatta
     5521 ctggcgtcaa cttttgaatg ctaaattaat tactcaaaga aaatttgata atttgactaa
     5581 agcagaaaga ggtgggcttt ctgaattaga taaagccggg ttcattaaaa gacaattggt
     5641 cgaaactaga caaattacta aacatgttgc ccaaatttta gattcccgta tgaacactaa
     5701 gtatgacgaa aatgataagt taatacgtga ggttaaagtc attactttaa aatcaaaact
     5761 tgtctctgat ttcagaaagg atttccaatt ctataaagtt agagaaatta ataattatca
     5821 tcatgctcat gatgcatatt tgaatgctgt agttggaact gctttaatca agaaataccc
     5881 taaattagaa tctgaatttg tatatggtga ttacaaagtc tatgatgtta gaaagatgat
     5941 tgctaaatca gaacaagaaa ttggtaaagc tacagctaaa tacttctttt actctaacat
     6001 tatgaatttc tttaaaacag aaattacttt ggcaaacggt gaaattagaa aaagacctct
     6061 tattgaaaca aatggtgaga ctggagagat agtttgggac aaagggcgtg atttcgctac
     6121 tgttagaaaa gttttatcaa tgccacaagt taacattgta aagaaaacag aggttcaaac
     6181 tggtggtttc tcaaaagaaa gtattttgcc taaaagaaat agtgataaat tgattgccag
     6241 aaaaaaggat tgggatccaa agaaatatgg tggtttcgac tcaccaaccg tagcctattc
     6301 tgttttggtt gtggcaaagg ttgaaaaggg taaaagtaaa aagcttaaat cagtaaaaga
     6361 acttttgggt attacaataa tggaaagaag ttcctttgaa aagaacccta ttgatttttt
     6421 ggaagctaaa ggttataagg aagtaaagaa ggacttaata atcaaattgc ctaaatattc
     6481 tttatttgaa ttagaaaatg ggagaaaaag aatgttggct tctgctggag aattgcaaaa
     6541 gggtaatgaa ttagcattgc cttccaaata tgttaacttc ttgtatttag cttcacacta
     6601 tgaaaagttg aaagggtcac cagaagataa cgagcaaaaa caattatttg ttgaacaaca
     6661 caaacactac ttagatgaga ttatagaaca aattagtgaa ttcagtaaaa gagtgatatt
     6721 agctgatgca aatttagata aagttttgtc agcctataac aaacatagag ataagccaat
     6781 tagagaacaa gcagaaaaca ttattcactt atttaccctt accaatttag gagcacctgc
     6841 tgctttcaag tattttgata caacaattga tcgtaaaaga tatacctcaa caaaagaagt
     6901 cttagacgcc accttaattc atcaatcaat cactggattg tatgagacaa gaattgattt
     6961 gtctcaattg ggtggtgatg aaggggctga tcctaagaag aaaagaaaag ttgatccaaa
     7021 gaaaaagcgt aaggtggatc ctaagaaaaa gagaaaggtt gactacaaag accatgacgg
     7081 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtgat aatctcttct
     7141 cgagtcatgt aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta
     7201 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt
     7261 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc
     7321 gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag
     7381 gctttaattt gcggccggta ctgcggtgtg aaataccgca cagatgcgta aggagaaaat
     7441 accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     7501 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     7561 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     7621 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     7681 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     7741 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     7801 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     7861 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc
     7921 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     7981 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     8041 cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag
     8101 ctccaccgcg gtggcggccg ctctagaact agtggatccc ccgggctgca ggaattcgat
     8161 atcaagcttt ctttgaaaag ataatgtatg attatgcttt cactcatatt tatacagaaa
     8221 cttgatgttt tctttcgagt atatacaagg tgattacatg tacgtttgaa gtacaactct
     8281 agattttgta gtgccctctt gggctagcgg taaaggtgcg cattttttca caccctacaa
     8341 tgttctgttc aaaagatttt ggtcaaacgc tgtagaagtg aaagttggtg cgcatgtttc
     8401 ggcgttcgaa acttctccgc agtgaaagat aaatgatcct agatgcgaga tatcctcatg
     8461 cctaggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     8521 aaagtggcac cgagtcggtg gtgctttttt tgttttttat gtctaccgtc gacctcgagg
     8581 gggggcccgg tacccagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa
     8641 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     8701 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     8761 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     8821 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     8881 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     8941 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     9001 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     9061 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     9121 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     9181 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     9241 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     9301 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     9361 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     9421 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     9481 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     9541 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     9601 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     9661 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     9721 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     9781 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     9841 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     9901 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     9961 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
    10021 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
    10081 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
    10141 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
    10201 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
    10261 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
    10321 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
    10381 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
    10441 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
    10501 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
    10561 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
    10621 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
    10681 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
    10741 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggg
    10801 tccttttcat cacgtgctat aaaaataatt ataatttaaa ttttttaata taaatatata
    10861 aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga
    10921 tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct cttcctgctc
    10981 tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aagaccacac acgaaaatcc
    11041 tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc tgcttttctt
    11101 gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac ctttgtttat
    11161 ttttttttct tcattccgta actcttctac cttctttatt tactttctaa aatccaaata
    11221 caaaacataa aaataaataa acacagagta aattcccaaa ttattccatc attaaaagat
    11281 acgaggcgcg tgtaagttac aggcaagcga tccgtctaag aaaccattat tatcatgaca
    11341 ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
