LOCUS       sequence-193437-       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 ARS911b sequence 
            GTAATATTGTCTTGTTTCCC in yeast chromosome 9..
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-ARS911b
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     371..388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    505..526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(541..546,547..564,565..571)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    579..595
                     /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)."
     primer_bind     584..606
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     603..619
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     603..619
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     638..658
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        640..658
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        885..960
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1046..1750
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1760..5863
                     /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"
     CDS             5876..5896
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5912..6159
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6177..6196)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6178..6196)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6206..6223)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6206..6222)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6215..6237)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6363..6818
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6450..6469)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6660..6681
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6949..7752)
                     /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"
     promoter        complement(7753..7968)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7992..8011)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8111..8133
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8171..8189)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      8230..9572
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    8562..8609
                     /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)."
     promoter        9599..9703
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9704..9772,9773..10564)
                     /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     complement(9922..9941)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(10735..11106,1..217)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
       61 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
      241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
      301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
      361 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
      421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
      481 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      541 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      601 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
      661 acaaaagctg gagctagtat actctttctt caacaattaa atactctcgg tagccaagtt
      721 ggtttaaggc gcaagactgt aatttatcac tacgaaatct tgagatcggg cgttcgactc
      781 gcccccggga gagatggccg gcatggtccc agcctcctcg ctggcgccgg ctgggcaaca
      841 ccttcgggtg gcgaatggga ctttgtaata ttgtcttgtt tcccgtttta gagctagaaa
      901 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
      961 tttttttatt ttttgtcact attgttatgt aaaatgccac ctctgacagt atggaacgca
     1021 aacttctgtc tagtggatag tcgacatcct tttgttgttt ccgggtgtac aatatggact
     1081 tcctcttttc tggcaaccaa acccatacat cgggattcct ataatacctt cgttggtctc
     1141 cctaacatgt aggtggcgga ggggagatat acaatagaac agataccaga caagacataa
     1201 tgggctaaac aagactacac caattacact gcctcattga tggtggtaca taacgaacta
     1261 atactgtagc cctagacttg atagccatca tcatatcgaa gtttcactac cctttttcca
     1321 tttgccatct attgaagtaa taataggcgc atgcaacttc ttttcttttt ttttcttttc
     1381 tctctccccc gttgttgtct caccatatcc gcaatgataa aaaaatgatg gaagacacta
     1441 aaggaaaaaa ttaacgacaa agacagcacc aacagatgtc gttgttccag agctgatgag
     1501 gggtatctcg aagcacacga aactttttcc ttccttcatt cacgcacact actctctaat
     1561 gagcaacggt atacggcctt ccttccagtt acttgaattt gaaataaaaa aaagtttgct
     1621 gtcttgctat caagtataaa tagacctgca attattaatc ttttgtttcc tcgtcattgt
     1681 tctcgttccc tttcttcctt gtttcttttt ctgcacaata tttcaagcta taccaagcat
     1741 acaatcaact tggaaaaaaa tggacaagaa gtactccatt gggctcgata tcggcacaaa
     1801 cagcgtcggt tgggccgtca ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt
     1861 tctgggcaat accgatcgcc acagcataaa gaagaacctc attggcgccc tcctgttcga
     1921 ctccggggag acggccgaag ccacgcggct caaaagaaca gcacggcgca gatatacccg
     1981 cagaaagaat cggatctgct acctgcagga gatctttagt aatgagatgg ctaaggtgga
     2041 tgactctttc ttccataggc tggaggagtc ctttttggtg gaggaggata aaaagaacga
     2101 gcgccaccca atctttggca atatcgtgga cgaggtggcg taccatgaaa agtacccaac
     2161 catatatcat ctgaggaaga agcttgtaga cagtactgat aaggctgact tgcggttgat
     2221 ctatctcgcg ctggcgcata tgatcaaatt tcggggacac ttcctcatcg agggggacct
     2281 gaacccagac aacagcgatg tcgacaaact ctttatccaa ctggttcaga cttacaatca
     2341 gcttttcgaa gagaacccga tcaacgcatc cggagttgac gccaaagcaa tcctgagcgc
     2401 taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca cagctccctg gggagaagaa
     2461 gaacggcctg tttggtaatc ttatcgccct gtcactcggg ctgaccccca actttaaatc
     2521 taacttcgac ctggccgaag atgccaagct tcaactgagc aaagacacct acgatgatga
     2581 tctcgacaat ctgctggccc agatcggcga ccagtacgca gacctttttt tggcggcaaa
     2641 gaacctgtca gacgccattc tgctgagtga tattctgcga gtgaacacgg agatcaccaa
     2701 agctccgctg agcgctagta tgatcaagcg ctatgatgag caccaccaag acttgacttt
     2761 gctgaaggcc cttgtcagac agcaactgcc tgagaagtac aaggaaattt tcttcgatca
     2821 gtctaaaaat ggctacgccg gatacattga cggcggagca agccaggagg aattttacaa
     2881 atttattaag cccatcttgg aaaaaatgga cggcaccgag gagctgctgg taaagcttaa
     2941 cagagaagat ctgttgcgca aacagcgcac tttcgacaat ggaagcatcc cccaccagat
     3001 tcacctgggc gaactgcacg ctatcctcag gcggcaagag gatttctacc cctttttgaa
     3061 agataacagg gaaaagattg agaaaatcct cacatttcgg ataccctact atgtaggccc
     3121 cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc aaatcagaag agaccatcac
     3181 tccctggaac ttcgaggaag tcgtggataa gggggcctct gcccagtcct tcatcgaaag
     3241 gatgactaac tttgataaaa atctgcctaa cgaaaaggtg cttcctaaac actctctgct
     3301 gtacgagtac ttcacagttt ataacgagct caccaaggtc aaatacgtca cagaagggat
     3361 gagaaagcca gcattcctgt ctggagagca gaagaaagct atcgtggacc tcctcttcaa
     3421 gacgaaccgg aaagttaccg tgaaacagct caaagaagac tatttcaaaa agattgaatg
     3481 tttcgactct gttgaaatca gcggagtgga ggatcgcttc aacgcatccc tgggaacgta
     3541 tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg gacaatgagg agaacgagga
     3601 cattcttgag gacattgtcc tcacccttac gttgtttgaa gatagggaga tgattgaaga
     3661 acgcttgaaa acttacgctc atctcttcga cgacaaagtc atgaaacagc tcaagaggcg
     3721 ccgatataca ggatgggggc ggctgtcaag aaaactgatc aatgggatcc gagacaagca
     3781 gagtggaaag acaatcctgg attttcttaa gtccgatgga tttgccaacc ggaacttcat
     3841 gcagttgatc catgatgact ctctcacctt taaggaggac atccagaaag cacaagtttc
     3901 tggccagggg gacagtcttc acgagcacat cgctaatctt gcaggtagcc cagctatcaa
     3961 aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc gtcaaagtaa tgggaaggca
     4021 taagcccgag aatatcgtta tcgagatggc ccgagagaac caaactaccc agaagggaca
     4081 gaagaacagt agggaaagga tgaagaggat tgaagagggt ataaaagaac tggggtccca
     4141 aatccttaag gaacacccag ttgaaaacac ccagcttcag aatgagaagc tctacctgta
     4201 ctacctgcag aacggcaggg acatgtacgt ggatcaggaa ctggacatca atcggctctc
     4261 cgactacgac gtggatcata tcgtgcccca gtcttttctc aaagatgatt ctattgataa
     4321 taaagtgttg acaagatccg ataaaaatag agggaagagt gataacgtcc cctcagaaga
     4381 agttgtcaag aaaatgaaaa attattggcg gcagctgctg aacgccaaac tgatcacaca
     4441 acggaagttc gataatctga ctaaggctga acgaggtggc ctgtctgagt tggataaagc
     4501 cggcttcatc aaaaggcagc ttgttgagac acgccagatc accaagcacg tggcccaaat
     4561 tctcgattca cgcatgaaca ccaagtacga tgaaaatgac aaactgattc gagaggtgaa
     4621 agttattact ctgaagtcta agctggtctc agatttcaga aaggactttc agttttataa
     4681 ggtgagagag atcaacaatt accaccatgc gcatgatgcc tacctgaatg cagtggtagg
     4741 cactgcactt atcaaaaaat atcccaagct tgaatctgaa tttgtttacg gagactataa
     4801 agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag gaaataggca aggccaccgc
     4861 taagtacttc ttttacagca atattatgaa ttttttcaag accgagatta cactggccaa
     4921 tggagagatt cggaagcgac cacttatcga aacaaacgga gaaacaggag aaatcgtgtg
     4981 ggacaagggt agggatttcg cgacagtccg gaaggtcctg tccatgccgc aggtgaacat
     5041 cgttaaaaag accgaagtac agaccggagg cttctccaag gaaagtatcc tcccgaaaag
     5101 gaacagcgac aagctgatcg cacgcaaaaa agattgggac cccaagaaat acggcggatt
     5161 cgattctcct acagtcgctt acagtgtact ggttgtggcc aaagtggaga aagggaagtc
     5221 taaaaaactc aaaagcgtca aggaactgct gggcatcaca atcatggagc gatcaagctt
     5281 cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat aaagaggtca aaaaagacct
     5341 catcattaag cttcccaagt actctctctt tgagcttgaa aacggccgga aacgaatgct
     5401 cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca ctgccctcta aatacgttaa
     5461 tttcttgtat ctggccagcc actatgaaaa gctcaaaggg tctcccgaag ataatgagca
     5521 gaagcagctg ttcgtggaac aacacaaaca ctaccttgat gagatcatcg agcaaataag
     5581 cgaattctcc aaaagagtga tcctcgccga cgctaacctc gataaggtgc tttctgctta
     5641 caataagcac agggataagc ccatcaggga gcaggcagaa aacattatct acttgtttac
     5701 tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc gacaccacca tagacagaaa
     5761 gcggtacacc tctacaaagg aggtcctgga cgccacactg attcatcagt caattacggg
     5821 gctctatgaa acaagaatcg acctctctca gctcggtgga gacagcaggg ctgaccccaa
     5881 gaagaagagg aaggtgtgat ctcttctcga gtcatgtaat tagttatgtc acgcttacgt
     5941 tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc
     6001 taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa
     6061 atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg
     6121 cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg gccggtaccc aattcgccct
     6181 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     6241 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     6301 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     6361 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     6421 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     6481 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     6541 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     6601 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     6661 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     6721 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     6781 taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
     6841 acgcatctgt gcggtatttc acaccgcata gggtaataac tgatataatt aaattgaagc
     6901 tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg
     6961 gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt
     7021 agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga
     7081 gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc
     7141 cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg
     7201 agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt
     7261 atattctcca gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct
     7321 aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac
     7381 cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta
     7441 ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt
     7501 gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa
     7561 gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag
     7621 taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat
     7681 gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt
     7741 agctttcgac atgatttatc ttcgtttcct gcaggttttt gttctgtgca gttgggttaa
     7801 gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag
     7861 tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca
     7921 aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa ggtggtatgg
     7981 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     8041 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     8101 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     8161 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     8221 tcttagtatg atccaatatc aaaggaaatg atagcattga aggatgagac taatccaatt
     8281 gaggagtggc agcatataga acagctaaag ggtagtgctg aaggaagcat acgatacccc
     8341 gcatggaatg ggataatatc acaggaggta ctagactacc tttcatccta cataaataga
     8401 cgcatataag tacgcattta agcataaaca cgcactatgc cgttcttctc atgtatatat
     8461 atatacaggc aacacgcaga tataggtgcg acgtgaacag tgagctgtat gtgcgcagct
     8521 cgcgttgcat tttcggaagc gctcgttttc ggaaacgctt tgaagttcct attccgaagt
     8581 tcctattctc tagaaagtat aggaacttca gagcgctttt gaaaaccaaa agcgctctga
     8641 agacgcactt tcaaaaaacc aaaaacgcac cggactgtaa cgagctacta aaatattgcg
     8701 aataccgctt ccacaaacat tgctcaaaag tatctctttg ctatatatct ctgtgctata
     8761 tccctatata acctacccat ccacctttcg ctccttgaac ttgcatctaa actcgacctc
     8821 tacatttttt atgtttatct ctagtattac tctttagaca aaaaaattgt agtaagaact
     8881 attcatagag tgaatcgaaa acaatacgaa aatgtaaaca tttcctatac gtagtatata
     8941 gagacaaaat agaagaaacc gttcataatt ttctgaccaa tgaagaatca tcaacgctat
     9001 cactttctgt tcacaaagta tgcgcaatcc acatcggtat agaatataat cggggatgcc
     9061 tttatcttga aaaaatgcac ccgcagcttc gctagtaatc agtaaacgcg ggaagtggag
     9121 tcaggctttt tttatggaag agaaaataga caccaaagta gccttcttct aaccttaacg
     9181 gacctacagt gcaaaaagtt atcaagagac tgcattatag agcgcacaaa ggagaaaaaa
     9241 agtaatctaa gatgctttgt tagaaaaata gcgctctcgg gatgcatttt tgtagaacaa
     9301 aaaagaagta tagattcttt gttggtaaaa tagcgctctc gcgttgcatt tctgttctgt
     9361 aaaaatgcag ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc atttttgttt
     9421 tacaaaaatg aagcacagat tcttcgttgg taaaatagcg ctttcgcgtt gcatttctgt
     9481 tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta gcgctctcgc gttgcatttt
     9541 tgttctacaa aatgaagcac agatgcttcg ttcaggtggc acttttcggg gaaatgtgcg
     9601 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     9661 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     9721 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     9781 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     9841 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     9901 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     9961 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
    10021 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
    10081 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
    10141 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
    10201 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
    10261 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
    10321 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
    10381 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
    10441 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
    10501 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
    10561 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
    10621 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
    10681 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
    10741 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
    10801 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
    10861 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
    10921 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
    10981 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
    11041 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
    11101 cgaact
//
