LOCUS       sequence-193434-       11106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-one CRISPR plasmid for integrating, deleting, or replacing a 
            gene in S. cerevisiae. High-copy, URA3 plasmid contains human 
            codon-optimized Cas9 and a guide RNA targeting CAN1y sequence 
            GATACGTTCTCTATGGAGGA in yeast chromosome 6..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11106)
  AUTHORS   Reider Apel A, d'Espaux L, Wehrs M, Sachs D, Li RA, Tong GJ, Garber 
            M, Nnadi O, Zhuang W, Hillson NJ, Keasling JD, Mukhopadhyay A
  TITLE     A Cas9-based toolkit to program gene expression in Saccharomyces 
            cerevisiae.
  JOURNAL   Nucleic Acids Res. 2017 Jan 9;45(1):496-508. doi: 
            10.1093/nar/gkw1023. Epub 2016 Nov 28.
  PUBMED    27899650
REFERENCE   2  (bases 1 to 11106)
  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: 
            "10.1093/nar/gkw1023"; journalName: "Nucleic Acids Res"; date: 
            "2017-01-9- 9"; volume: "45"; issue: "1"; pages: "496-508"
COMMENT     Sequence Label: p426_Cas9_gRNA-CAN1y
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..40)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(194..782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(274..293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(953..1744,1745..1813))
                     /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     1576..1595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1814..1918)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1945..3287
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(2908..2955)
                     /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)."
     primer_bind     3328..3346
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3384..3406)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3506..3525
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3549..3764
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3765..4568
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(4699..5154)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4836..4857)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5048..5067
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5280..5302
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5294..5311
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5295..5311
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5321..5340
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5321..5339
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(5358..5605)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(5621..5641)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(5654..9757)
                     /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
                     FLVEEDKKNERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIYLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     promoter        complement(9767..10471)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     misc_RNA        complement(10557..10632)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(10859..10879)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(10859..10877)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10898..10914)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10898..10914)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10911..10933)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10922..10938
                     /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(10946..10952,10953..10970,10971..10976))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10991..11012
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
       61 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      121 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      181 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      241 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      301 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      361 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      421 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      481 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      541 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      601 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
      661 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      721 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      781 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      841 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      901 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
      961 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1021 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1081 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1141 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1201 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1261 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1321 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1381 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1441 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1501 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1561 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1621 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1681 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1741 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1801 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1861 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1921 catttccccg aaaagtgcca cctgaacgaa gcatctgtgc ttcattttgt agaacaaaaa
     1981 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     2041 aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac
     2101 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag
     2161 aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt
     2221 tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt
     2281 ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt
     2341 aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc
     2401 acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca
     2461 tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag
     2521 cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata
     2581 tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt
     2641 cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg
     2701 tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata
     2761 gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca
     2821 atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcttcag
     2881 agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt
     2941 cggaatagga acttcaaagc gtttccgaaa acgagcgctt ccgaaaatgc aacgcgagct
     3001 gcgcacatac agctcactgt tcacgtcgca cctatatctg cgtgttgcct gtatatatat
     3061 atacatgaga agaacggcat agtgcgtgtt tatgcttaaa tgcgtactta tatgcgtcta
     3121 tttatgtagg atgaaaggta gtctagtacc tcctgtgata ttatcccatt ccatgcgggg
     3181 tatcgtatgc ttccttcagc actacccttt agctgttcta tatgctgcca ctcctcaatt
     3241 ggattagtct catccttcaa tgctatcatt tcctttgata ttggatcata ctaagaaacc
     3301 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     3361 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     3421 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     3481 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     3541 accacctttt caattcatca tttttttttt attctttttt ttgatttcgg tttccttgaa
     3601 atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta
     3661 gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa
     3721 cccaactgca cagaacaaaa acctgcagga aacgaagata aatcatgtcg aaagctacat
     3781 ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc
     3841 acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg
     3901 agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga
     3961 ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt
     4021 ttttactctt cgaagacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact
     4081 ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg
     4141 gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag
     4201 gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta
     4261 agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa
     4321 gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt
     4381 tagatgacaa gggagacgca ttgggtcaac agtatagaac cgtggatgat gtggtctcta
     4441 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg
     4501 tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc
     4561 aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt
     4621 caatttaatt atatcagtta ttaccctatg cggtgtgaaa taccgcacag atgcgtaagg
     4681 agaaaatacc gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4741 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4801 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4861 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4921 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4981 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     5041 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     5101 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc
     5161 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5221 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     5281 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg
     5341 cgaattgggt accggccgca aattaaagcc ttcgagcgtc ccaaaacctt ctcaagcaag
     5401 gttttcagta taatgttaca tgcgtacacg cgtctgtaca gaaaaaaaag aaaaatttga
     5461 aatataaata acgttcttaa tactaacata actataaaaa aataaatagg gacctagact
     5521 tcaggttgtc taactccttc cttttcggtt agagcggatg tggggggagg gcgtgaacgt
     5581 aagcgtgaca taactaatta catgactcga gaagagatca caccttcctc ttcttcttgg
     5641 ggtcagccct gctgtctcca ccgagctgag agaggtcgat tcttgtttca tagagccccg
     5701 taattgactg atgaatcagt gtggcgtcca ggacctcctt tgtagaggtg taccgctttc
     5761 tgtctatggt ggtgtcgaag tacttgaagg ctgcaggcgc gcccaagttg gtcagagtaa
     5821 acaagtagat aatgttttct gcctgctccc tgatgggctt atccctgtgc ttattgtaag
     5881 cagaaagcac cttatcgagg ttagcgtcgg cgaggatcac tcttttggag aattcgctta
     5941 tttgctcgat gatctcatca aggtagtgtt tgtgttgttc cacgaacagc tgcttctgct
     6001 cattatcttc gggagaccct ttgagctttt catagtggct ggccagatac aagaaattaa
     6061 cgtatttaga gggcagtgcc agctcgttac ctttctgcag ctcgcccgca ctagcgagca
     6121 ttcgtttccg gccgttttca agctcaaaga gagagtactt gggaagctta atgatgaggt
     6181 cttttttgac ctctttatat cctttcgcct cgagaaagtc gatggggttt ttttcgaagc
     6241 ttgatcgctc catgattgtg atgcccagca gttccttgac gcttttgagt tttttagact
     6301 tccctttctc cactttggcc acaaccagta cactgtaagc gactgtagga gaatcgaatc
     6361 cgccgtattt cttggggtcc caatcttttt tgcgtgcgat cagcttgtcg ctgttccttt
     6421 tcgggaggat actttccttg gagaagcctc cggtctgtac ttcggtcttt ttaacgatgt
     6481 tcacctgcgg catggacagg accttccgga ctgtcgcgaa atccctaccc ttgtcccaca
     6541 cgatttctcc tgtttctccg tttgtttcga taagtggtcg cttccgaatc tctccattgg
     6601 ccagtgtaat ctcggtcttg aaaaaattca taatattgct gtaaaagaag tacttagcgg
     6661 tggccttgcc tatttcctgc tcagactttg cgatcatttt cctaacatcg tacactttat
     6721 agtctccgta aacaaattca gattcaagct tgggatattt tttgataagt gcagtgccta
     6781 ccactgcatt caggtaggca tcatgcgcat ggtggtaatt gttgatctct ctcaccttat
     6841 aaaactgaaa gtcctttctg aaatctgaga ccagcttaga cttcagagta ataactttca
     6901 cctctcgaat cagtttgtca ttttcatcgt acttggtgtt catgcgtgaa tcgagaattt
     6961 gggccacgtg cttggtgatc tggcgtgtct caacaagctg ccttttgatg aagccggctt
     7021 tatccaactc agacaggcca cctcgttcag ccttagtcag attatcgaac ttccgttgtg
     7081 tgatcagttt ggcgttcagc agctgccgcc aataattttt cattttcttg acaacttctt
     7141 ctgaggggac gttatcactc ttccctctat ttttatcgga tcttgtcaac actttattat
     7201 caatagaatc atctttgaga aaagactggg gcacgatatg atccacgtcg tagtcggaga
     7261 gccgattgat gtccagttcc tgatccacgt acatgtccct gccgttctgc aggtagtaca
     7321 ggtagagctt ctcattctga agctgggtgt tttcaactgg gtgttcctta aggatttggg
     7381 accccagttc ttttataccc tcttcaatcc tcttcatcct ttccctactg ttcttctgtc
     7441 ccttctgggt agtttggttc tctcgggcca tctcgataac gatattctcg ggcttatgcc
     7501 ttcccattac tttgacgagt tcatccacga ccttaacggt ctgcagtatt ccctttttga
     7561 tagctgggct acctgcaaga ttagcgatgt gctcgtgaag actgtccccc tggccagaaa
     7621 cttgtgcttt ctggatgtcc tccttaaagg tgagagagtc atcatggatc aactgcatga
     7681 agttccggtt ggcaaatcca tcggacttaa gaaaatccag gattgtcttt ccactctgct
     7741 tgtctcggat cccattgatc agttttcttg acagccgccc ccatcctgta tatcggcgcc
     7801 tcttgagctg tttcatgact ttgtcgtcga agagatgagc gtaagttttc aagcgttctt
     7861 caatcatctc cctatcttca aacaacgtaa gggtgaggac aatgtcctca agaatgtcct
     7921 cgttctcctc attgtccagg aagtccttgt ctttaatgat tttcaggaga tcgtgatacg
     7981 ttcccaggga tgcgttgaag cgatcctcca ctccgctgat ttcaacagag tcgaaacatt
     8041 caatcttttt gaaatagtct tctttgagct gtttcacggt aactttccgg ttcgtcttga
     8101 agaggaggtc cacgatagct ttcttctgct ctccagacag gaatgctggc tttctcatcc
     8161 cttctgtgac gtatttgacc ttggtgagct cgttataaac tgtgaagtac tcgtacagca
     8221 gagagtgttt aggaagcacc ttttcgttag gcagattttt atcaaagtta gtcatccttt
     8281 cgatgaagga ctgggcagag gcccccttat ccacgacttc ctcgaagttc cagggagtga
     8341 tggtctcttc tgatttgcga gtcatccacg cgaatctgga atttccccgg gcgagggggc
     8401 ctacatagta gggtatccga aatgtgagga ttttctcaat cttttccctg ttatctttca
     8461 aaaaggggta gaaatcctct tgccgcctga ggatagcgtg cagttcgccc aggtgaatct
     8521 ggtgggggat gcttccattg tcgaaagtgc gctgtttgcg caacagatct tctctgttaa
     8581 gctttaccag cagctcctcg gtgccgtcca ttttttccaa gatgggctta ataaatttgt
     8641 aaaattcctc ctggcttgct ccgccgtcaa tgtatccggc gtagccattt ttagactgat
     8701 cgaagaaaat ttccttgtac ttctcaggca gttgctgtct gacaagggcc ttcagcaaag
     8761 tcaagtcttg gtggtgctca tcatagcgct tgatcatact agcgctcagc ggagctttgg
     8821 tgatctccgt gttcactcgc agaatatcac tcagcagaat ggcgtctgac aggttctttg
     8881 ccgccaaaaa aaggtctgcg tactggtcgc cgatctgggc cagcagattg tcgagatcat
     8941 catcgtaggt gtctttgctc agttgaagct tggcatcttc ggccaggtcg aagttagatt
     9001 taaagttggg ggtcagcccg agtgacaggg cgataagatt accaaacagg ccgttcttct
     9061 tctccccagg gagctgtgcg atgaggtttt cgagccgccg ggatttggac agcctagcgc
     9121 tcaggattgc tttggcgtca actccggatg cgttgatcgg gttctcttcg aaaagctgat
     9181 tgtaagtctg aaccagttgg ataaagagtt tgtcgacatc gctgttgtct gggttcaggt
     9241 ccccctcgat gaggaagtgt ccccgaaatt tgatcatatg cgccagcgcg agatagatca
     9301 accgcaagtc agccttatca gtactgtcta caagcttctt cctcagatga tatatggttg
     9361 ggtacttttc atggtacgcc acctcgtcca cgatattgcc aaagattggg tggcgctcgt
     9421 tctttttatc ctcctccacc aaaaaggact cctccagcct atggaagaaa gagtcatcca
     9481 ccttagccat ctcattacta aagatctcct gcaggtagca gatccgattc tttctgcggg
     9541 tatatctgcg ccgtgctgtt cttttgagcc gcgtggcttc ggccgtctcc ccggagtcga
     9601 acaggagggc gccaatgagg ttcttcttta tgctgtggcg atcggtattg cccagaactt
     9661 tgaatttttt gctcggcacc ttgtactcgt ccgtaatgac ggcccaaccg acgctgtttg
     9721 tgccgatatc gagcccaatg gagtacttct tgtccatttt tttccaagtt gattgtatgc
     9781 ttggtatagc ttgaaatatt gtgcagaaaa agaaacaagg aagaaaggga acgagaacaa
     9841 tgacgaggaa acaaaagatt aataattgca ggtctattta tacttgatag caagacagca
     9901 aacttttttt tatttcaaat tcaagtaact ggaaggaagg ccgtataccg ttgctcatta
     9961 gagagtagtg tgcgtgaatg aaggaaggaa aaagtttcgt gtgcttcgag atacccctca
    10021 tcagctctgg aacaacgaca tctgttggtg ctgtctttgt cgttaatttt ttcctttagt
    10081 gtcttccatc atttttttgt cattgcggat atggtgagac aacaacgggg gagagagaaa
    10141 agaaaaaaaa agaaaagaag ttgcatgcgc ctattattac ttcaatagat ggcaaatgga
    10201 aaaagggtag tgaaacttcg atatgatgat ggctatcaag tctagggcta cagtattagt
    10261 tcgttatgta ccaccatcaa tgaggcagtg taattggtgt agtcttgttt agcccattat
    10321 gtcttgtctg gtatctgttc tattgtatat ctcccctccg ccacctacat gttagggaga
    10381 ccaacgaagg tattatagga atcccgatgt atgggtttgg ttgccagaaa agaggaagtc
    10441 catattgtac acccggaaac aacaaaagga tgtcgactat ccactagaca gaagtttgcg
    10501 ttccatactg tcagaggtgg cattttacat aacaatagtg acaaaaaata aaaaaagcac
    10561 cgactcggtg ccactttttc aagttgataa cggactagcc ttattttaac ttgctatttc
    10621 tagctctaaa actcctccat agagaacgta tcaaagtccc attcgccacc cgaaggtgtt
    10681 gcccagccgg cgccagcgag gaggctggga ccatgccggc catctctccc gggggcgagt
    10741 cgaacgcccg atctcaagat ttcgtagtga taaattacag tcttgcgcct taaaccaact
    10801 tggctaccga gagtatttaa ttgttgaaga aagagtatac tagctccagc ttttgttccc
    10861 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
    10921 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
    10981 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
    11041 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
    11101 gtttgc
//
