LOCUS       sequence-193449-       11597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  p425_Cas9_gRNA-ARS1014a All-in-one CRISPR plasmid for integrating, 
            deleting, or replacing a gene in S. cerevisiae. High-copy, LEU2 
            plasmid contains human codon-optimized Cas9 and a guide RNA 
            targeting ARS1014a sequence TTATGTGCGTATTGCTTTCA in yeast chromosome
            10..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11597)
  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 11597)
  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: p425_Cas9_gRNA_LEU_1014a
FEATURES             Location/Qualifiers
     source          1..11597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     381..398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    515..536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(551..556,557..574,575..581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    589..605
                     /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     594..616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     613..629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     613..629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     648..668
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        650..668
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        895..970
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1056..1760
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1770..5873
                     /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             5886..5906
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5922..6169
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6187..6206)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6188..6206)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6216..6233)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6216..6232)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6225..6247)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6373..6828
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6460..6479)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6670..6691
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6959..8053)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(8054..8463)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(8493..8512)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8612..8634
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8672..8690)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      8731..10073
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    9063..9110
                     /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        10100..10204
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10205..10273,10274..11065)
                     /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(10423..10442)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(11236..11597,1..227)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
       61 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      481 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      541 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      601 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
      661 actaaaggga acaaaagctg gagctagtat actctttctt caacaattaa atactctcgg
      721 tagccaagtt ggtttaaggc gcaagactgt aatttatcac tacgaaatct tgagatcggg
      781 cgttcgactc gcccccggga gagatggccg gcatggtccc agcctcctcg ctggcgccgg
      841 ctgggcaaca ccttcgggtg gcgaatggga ctttttatgt gcgtattgct ttcagtttta
      901 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
      961 gagtcggtgc tttttttatt ttttgtcact attgttatgt aaaatgccac ctctgacagt
     1021 atggaacgca aacttctgtc tagtggatag tcgacatcct tttgttgttt ccgggtgtac
     1081 aatatggact tcctcttttc tggcaaccaa acccatacat cgggattcct ataatacctt
     1141 cgttggtctc cctaacatgt aggtggcgga ggggagatat acaatagaac agataccaga
     1201 caagacataa tgggctaaac aagactacac caattacact gcctcattga tggtggtaca
     1261 taacgaacta atactgtagc cctagacttg atagccatca tcatatcgaa gtttcactac
     1321 cctttttcca tttgccatct attgaagtaa taataggcgc atgcaacttc ttttcttttt
     1381 ttttcttttc tctctccccc gttgttgtct caccatatcc gcaatgacaa aaaaatgatg
     1441 gaagacacta aaggaaaaaa ttaacgacaa agacagcacc aacagatgtc gttgttccag
     1501 agctgatgag gggtatctcg aagcacacga aactttttcc ttccttcatt cacgcacact
     1561 actctctaat gagcaacggt atacggcctt ccttccagtt acttgaattt gaaataaaaa
     1621 aaagtttgct gtcttgctat caagtataaa tagacctgca attattaatc ttttgtttcc
     1681 tcgtcattgt tctcgttccc tttcttcctt gtttcttttt ctgcacaata tttcaagcta
     1741 taccaagcat acaatcaact tggaaaaaaa tggacaagaa gtactccatt gggctcgata
     1801 tcggcacaaa cagcgtcggt tgggccgtca ttacggacga gtacaaggtg ccgagcaaaa
     1861 aattcaaagt tctgggcaat accgatcgcc acagcataaa gaagaacctc attggcgccc
     1921 tcctgttcga ctccggggag acggccgaag ccacgcggct caaaagaaca gcacggcgca
     1981 gatatacccg cagaaagaat cggatctgct acctgcagga gatctttagt aatgagatgg
     2041 ctaaggtgga tgactctttc ttccataggc tggaggagtc ctttttggtg gaggaggata
     2101 aaaagaacga gcgccaccca atctttggca atatcgtgga cgaggtggcg taccatgaaa
     2161 agtacccaac catatatcat ctgaggaaga agcttgtaga cagtactgat aaggctgact
     2221 tgcggttgat ctatctcgcg ctggcgcata tgatcaaatt tcggggacac ttcctcatcg
     2281 agggggacct gaacccagac aacagcgatg tcgacaaact ctttatccaa ctggttcaga
     2341 cttacaatca gcttttcgaa gagaacccga tcaacgcatc cggagttgac gccaaagcaa
     2401 tcctgagcgc taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca cagctccctg
     2461 gggagaagaa gaacggcctg tttggtaatc ttatcgccct gtcactcggg ctgaccccca
     2521 actttaaatc taacttcgac ctggccgaag atgccaagct tcaactgagc aaagacacct
     2581 acgatgatga tctcgacaat ctgctggccc agatcggcga ccagtacgca gacctttttt
     2641 tggcggcaaa gaacctgtca gacgccattc tgctgagtga tattctgcga gtgaacacgg
     2701 agatcaccaa agctccgctg agcgctagta tgatcaagcg ctatgatgag caccaccaag
     2761 acttgacttt gctgaaggcc cttgtcagac agcaactgcc tgagaagtac aaggaaattt
     2821 tcttcgatca gtctaaaaat ggctacgccg gatacattga cggcggagca agccaggagg
     2881 aattttacaa atttattaag cccatcttgg aaaaaatgga cggcaccgag gagctgctgg
     2941 taaagcttaa cagagaagat ctgttgcgca aacagcgcac tttcgacaat ggaagcatcc
     3001 cccaccagat tcacctgggc gaactgcacg ctatcctcag gcggcaagag gatttctacc
     3061 cctttttgaa agataacagg gaaaagattg agaaaatcct cacatttcgg ataccctact
     3121 atgtaggccc cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc aaatcagaag
     3181 agaccatcac tccctggaac ttcgaggaag tcgtggataa gggggcctct gcccagtcct
     3241 tcatcgaaag gatgactaac tttgataaaa atctgcctaa cgaaaaggtg cttcctaaac
     3301 actctctgct gtacgagtac ttcacagttt ataacgagct caccaaggtc aaatacgtca
     3361 cagaagggat gagaaagcca gcattcctgt ctggagagca gaagaaagct atcgtggacc
     3421 tcctcttcaa gacgaaccgg aaagttaccg tgaaacagct caaagaagac tatttcaaaa
     3481 agattgaatg tttcgactct gttgaaatca gcggagtgga ggatcgcttc aacgcatccc
     3541 tgggaacgta tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg gacaatgagg
     3601 agaacgagga cattcttgag gacattgtcc tcacccttac gttgtttgaa gatagggaga
     3661 tgattgaaga acgcttgaaa acttacgctc atctcttcga cgacaaagtc atgaaacagc
     3721 tcaagaggcg ccgatataca ggatgggggc ggctgtcaag aaaactgatc aatgggatcc
     3781 gagacaagca gagtggaaag acaatcctgg attttcttaa gtccgatgga tttgccaacc
     3841 ggaacttcat gcagttgatc catgatgact ctctcacctt taaggaggac atccagaaag
     3901 cacaagtttc tggccagggg gacagtcttc acgagcacat cgctaatctt gcaggtagcc
     3961 cagctatcaa aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc gtcaaagtaa
     4021 tgggaaggca taagcccgag aatatcgtta tcgagatggc ccgagagaac caaactaccc
     4081 agaagggaca gaagaacagt agggaaagga tgaagaggat tgaagagggt ataaaagaac
     4141 tggggtccca aatccttaag gaacacccag ttgaaaacac ccagcttcag aatgagaagc
     4201 tctacctgta ctacctgcag aacggcaggg acatgtacgt ggatcaggaa ctggacatca
     4261 atcggctctc cgactacgac gtggatcata tcgtgcccca gtcttttctc aaagatgatt
     4321 ctattgataa taaagtgttg acaagatccg ataaaaatag agggaagagt gataacgtcc
     4381 cctcagaaga agttgtcaag aaaatgaaaa attattggcg gcagctgctg aacgccaaac
     4441 tgatcacaca acggaagttc gataatctga ctaaggctga acgaggtggc ctgtctgagt
     4501 tggataaagc cggcttcatc aaaaggcagc ttgttgagac acgccagatc accaagcacg
     4561 tggcccaaat tctcgattca cgcatgaaca ccaagtacga tgaaaatgac aaactgattc
     4621 gagaggtgaa agttattact ctgaagtcta agctggtctc agatttcaga aaggactttc
     4681 agttttataa ggtgagagag atcaacaatt accaccatgc gcatgatgcc tacctgaatg
     4741 cagtggtagg cactgcactt atcaaaaaat atcccaagct tgaatctgaa tttgtttacg
     4801 gagactataa agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag gaaataggca
     4861 aggccaccgc taagtacttc ttttacagca atattatgaa ttttttcaag accgagatta
     4921 cactggccaa tggagagatt cggaagcgac cacttatcga aacaaacgga gaaacaggag
     4981 aaatcgtgtg ggacaagggt agggatttcg cgacagtccg gaaggtcctg tccatgccgc
     5041 aggtgaacat cgttaaaaag accgaagtac agaccggagg cttctccaag gaaagtatcc
     5101 tcccgaaaag gaacagcgac aagctgatcg cacgcaaaaa agattgggac cccaagaaat
     5161 acggcggatt cgattctcct acagtcgctt acagtgtact ggttgtggcc aaagtggaga
     5221 aagggaagtc taaaaaactc aaaagcgtca aggaactgct gggcatcaca atcatggagc
     5281 gatcaagctt cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat aaagaggtca
     5341 aaaaagacct catcattaag cttcccaagt actctctctt tgagcttgaa aacggccgga
     5401 aacgaatgct cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca ctgccctcta
     5461 aatacgttaa tttcttgtat ctggccagcc actatgaaaa gctcaaaggg tctcccgaag
     5521 ataatgagca gaagcagctg ttcgtggaac aacacaaaca ctaccttgat gagatcatcg
     5581 agcaaataag cgaattctcc aaaagagtga tcctcgccga cgctaacctc gataaggtgc
     5641 tttctgctta caataagcac agggataagc ccatcaggga gcaggcagaa aacattatct
     5701 acttgtttac tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc gacaccacca
     5761 tagacagaaa gcggtacacc tctacaaagg aggtcctgga cgccacactg attcatcagt
     5821 caattacggg gctctatgaa acaagaatcg acctctctca gctcggtgga gacagcaggg
     5881 ctgaccccaa gaagaagagg aaggtgtgat ctcttctcga gtcatgtaat tagttatgtc
     5941 acgcttacgt tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca
     6001 acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt
     6061 tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact
     6121 gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg gccggtaccc
     6181 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     6241 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     6301 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     6361 aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     6421 gcagcgtgac cactacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     6481 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     6541 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     6601 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     6661 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     6721 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6781 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     6841 ttttctcctt acgcatctgt gcggtatttc acaccgcata gggtaataac tgatataatt
     6901 aaattgaagc tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt
     6961 aagcaaggat tttcttaact tcttcggcga cagcatcacc gacttcggtg gtactgttgg
     7021 aaccacctaa atcaccagtt ctgatacctg catccaaaac ctttttaact gcatcttcaa
     7081 tggccttacc ttcttcaggc aagttcaatg acaatttcaa catcattgca gcagacaaga
     7141 tagtggcgat agggttgacc ttattctttg gcaaatctgg agcagaaccg tggcatggtt
     7201 cgtacaaacc aaatgcggtg ttcttgtctg gcaaagaggc caaggacgca gatggcaaca
     7261 aacccaagga acctgggata acggaggctt catcggagat gatatcacca aacatgttgc
     7321 tggtgattat aataccattt aggtgggttg ggttcttaac taggatcatg gcggcagaat
     7381 caatcaattg atgttgaacc ttcaatgtag ggaattcgtt cttgatggtt tcctccacag
     7441 tttttctcca taatcttgaa gaggccaaaa cattagcttt atccaaggac caaataggca
     7501 atggtggctc atgttgtagg gccatgaaag cggccattct tgtgattctt tgcacttctg
     7561 gaacggtgta ttgttcacta tcccaagcga caccatcacc atcgtcttcc tttctcttac
     7621 caaagtaaat acctcccact aattctctga caacaacgaa gtcagtacct ttagcaaatt
     7681 gtggcttgat tggagataag tctaaaagag agtcggatgc aaagttacat ggtcttaagt
     7741 tggcgtacaa ttgaagttct ttacggattt ttagtaaacc ttgttcaggt ctaacactac
     7801 cggtacccca tttaggacca cccacagcac ctaacaaaac ggcatcagcc ttcttggagg
     7861 cttccagcgc ctcatctgga agtggaacac ctgtagcatc gatagcagca ccaccaatta
     7921 aatgattttc gaaatcgaac ttgacattgg aacgaacatc agaaatagct ttaagaacct
     7981 taatggcttc ggctgtgatt tcttgaccaa cgtggtcacc tggcaaaacg acgatcttct
     8041 taggggcaga cattagaatg gtatatcctt gaaatatata tatatatatn tngctgaaat
     8101 gtaaaaggta agaaaagtta gaaagtaaga cgattgctaa ccacctattg gaaaaaacaa
     8161 taggtcctta aataatattg tcaacttcaa gtattgtgat gcaagcattt agtcatgaac
     8221 gcttctctat tctatatgaa aagccggttc cggcgctctc acctttcctt tttctcccaa
     8281 tttttcagtt gaaaaaggta tatgcgtcag gcgacctctg aaattaacaa aaaatttcca
     8341 gtcatcgaat ttgattctgt gcgatagcgc ccctgtgtgt tctcgttatg ttgaggaaaa
     8401 aaataatggt tgctaagaga ttcgaactct tgcatcttac gatacctgag tattcccaca
     8461 gtttgaaaag ctgtggtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     8521 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     8581 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     8641 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     8701 aatgtcatga taataatggt ttcttagtat gatccaatat caaaggaaat gatagcattg
     8761 aaggatgaga ctaatccaat tgaggagtgg cagcatatag aacagctaaa gggtagtgct
     8821 gaaggaagca tacgataccc cgcatggaat gggataatat cacaggaggt actagactac
     8881 ctttcatcct acataaatag acgcatataa gtacgcattt aagcataaac acgcactatg
     8941 ccgttcttct catgtatata tatatacagg caacacgcag atataggtgc gacgtgaaca
     9001 gtgagctgta tgtgcgcagc tcgcgttgca ttttcggaag cgctcgtttt cggaaacgct
     9061 ttgaagttcc tattccgaag ttcctattct ctagaaagta taggaacttc agagcgcttt
     9121 tgaaaaccaa aagcgctctg aagacgcact ttcaaaaaac caaaaacgca ccggactgta
     9181 acgagctact aaaatattgc gaataccgct tccacaaaca ttgctcaaaa gtatctcttt
     9241 gctatatatc tctgtgctat atccctatat aacctaccca tccacctttc gctccttgaa
     9301 cttgcatcta aactcgacct ctacattttt tatgtttatc tctagtatta ctctttagac
     9361 aaaaaaattg tagtaagaac tattcataga gtgaatcgaa aacaatacga aaatgtaaac
     9421 atttcctata cgtagtatat agagacaaaa tagaagaaac cgttcataat tttctgacca
     9481 atgaagaatc atcaacgcta tcactttctg ttcacaaagt atgcgcaatc cacatcggta
     9541 tagaatataa tcggggatgc ctttatcttg aaaaaatgca cccgcagctt cgctagtaat
     9601 cagtaaacgc gggaagtgga gtcaggcttt ttttatggaa gagaaaatag acaccaaagt
     9661 agccttcttc taaccttaac ggacctacag tgcaaaaagt tatcaagaga ctgcattata
     9721 gagcgcacaa aggagaaaaa aagtaatcta agatgctttg ttagaaaaat agcgctctcg
     9781 ggatgcattt ttgtagaaca aaaaagaagt atagattctt tgttggtaaa atagcgctct
     9841 cgcgttgcat ttctgttctg taaaaatgca gctcagattc tttgtttgaa aaattagcgc
     9901 tctcgcgttg catttttgtt ttacaaaaat gaagcacaga ttcttcgttg gtaaaatagc
     9961 gctttcgcgt tgcatttctg ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt
    10021 agcgctctcg cgttgcattt ttgttctaca aaatgaagca cagatgcttc gttcaggtgg
    10081 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
    10141 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
    10201 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
    10261 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
    10321 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
    10381 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
    10441 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
    10501 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
    10561 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
    10621 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
    10681 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
    10741 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
    10801 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
    10861 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
    10921 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
    10981 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
    11041 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
    11101 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
    11161 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
    11221 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
    11281 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
    11341 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
    11401 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
    11461 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
    11521 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
    11581 cttggagcga acgacct
//
