LOCUS       sequence-193450-       11034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  p426_Cas9_gRNA-ARS805a without ribozyme 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 ARS805a sequence TTATTTGAATGATATTTAGT in yeast 
            chromosome 8..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11034)
  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 11034)
  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-R-ARS805a
FEATURES             Location/Qualifiers
     source          1..11034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     261..280
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     514..531
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    648..669
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(684..689,690..707,708..714)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    722..738
                     /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     727..749
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     746..762
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     746..762
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     781..801
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        783..801
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        956..1031
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1117..1821
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1831..5934
                     /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             5947..5967
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5983..6230
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6248..6267)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6249..6267)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6277..6294)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6277..6293)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6286..6308)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6434..6889
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6521..6540)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6731..6752
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7020..7823)
                     /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(7824..8039)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(8063..8082)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8182..8204
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8242..8260)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      8301..9643
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    8633..8680
                     /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        9670..9774
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9775..9843,9844..10635)
                     /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(9993..10012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(10806..11034,1..360)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
       61 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      121 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      181 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      241 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      301 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      361 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      421 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      481 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      541 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      601 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      661 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      721 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
      781 gcaattaacc ctcactaaag ggaacaaaag ctggagctag tatactcttt cttcaacaat
      841 taaatactct cggtagccaa gttggtttaa ggcgcaagac tgtaatttat cactacgaaa
      901 tcttgagatc gggcgttcga ctcgcccccg ggagattatt tgaatgatat ttagtgtttt
      961 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
     1021 cgagtcggtg ctttttttat tttttgtcac tattgttatg taaaatgcca cctctgacag
     1081 tatggaacgc aaacttctgt ctagtggata gtcgacatcc ttttgttgtt tccgggtgta
     1141 caatatggac ttcctctttt ctggcaacca aacccataca tcgggattcc tataatacct
     1201 tcgttggtct ccctaacatg taggtggcgg aggggagata tacaatagaa cagataccag
     1261 acaagacata atgggctaaa caagactaca ccaattacac tgcctcattg atggtggtac
     1321 ataacgaact aatactgtag ccctagactt gatagccatc atcatatcga agtttcacta
     1381 ccctttttcc atttgccatc tattgaagta ataataggcg catgcaactt cttttctttt
     1441 tttttctttt ctctctcccc cgttgttgtc tcaccatatc cgcaatgaca aaaaaatgat
     1501 ggaagacact aaaggaaaaa attaacgaca aagacagcac caacagatgt cgttgttcca
     1561 gagctgatga ggggtatctc gaagcacacg aaactttttc cttccttcat tcacgcacac
     1621 tactctctaa tgagcaacgg tatacggcct tccttccagt tacttgaatt tgaaataaaa
     1681 aaaagtttgc tgtcttgcta tcaagtataa atagacctgc aattattaat cttttgtttc
     1741 ctcgtcattg ttctcgttcc ctttcttcct tgtttctttt tctgcacaat atttcaagct
     1801 ataccaagca tacaatcaac ttggaaaaaa atggacaaga agtactccat tgggctcgat
     1861 atcggcacaa acagcgtcgg ttgggccgtc attacggacg agtacaaggt gccgagcaaa
     1921 aaattcaaag ttctgggcaa taccgatcgc cacagcataa agaagaacct cattggcgcc
     1981 ctcctgttcg actccgggga gacggccgaa gccacgcggc tcaaaagaac agcacggcgc
     2041 agatataccc gcagaaagaa tcggatctgc tacctgcagg agatctttag taatgagatg
     2101 gctaaggtgg atgactcttt cttccatagg ctggaggagt cctttttggt ggaggaggat
     2161 aaaaagaacg agcgccaccc aatctttggc aatatcgtgg acgaggtggc gtaccatgaa
     2221 aagtacccaa ccatatatca tctgaggaag aagcttgtag acagtactga taaggctgac
     2281 ttgcggttga tctatctcgc gctggcgcat atgatcaaat ttcggggaca cttcctcatc
     2341 gagggggacc tgaacccaga caacagcgat gtcgacaaac tctttatcca actggttcag
     2401 acttacaatc agcttttcga agagaacccg atcaacgcat ccggagttga cgccaaagca
     2461 atcctgagcg ctaggctgtc caaatcccgg cggctcgaaa acctcatcgc acagctccct
     2521 ggggagaaga agaacggcct gtttggtaat cttatcgccc tgtcactcgg gctgaccccc
     2581 aactttaaat ctaacttcga cctggccgaa gatgccaagc ttcaactgag caaagacacc
     2641 tacgatgatg atctcgacaa tctgctggcc cagatcggcg accagtacgc agaccttttt
     2701 ttggcggcaa agaacctgtc agacgccatt ctgctgagtg atattctgcg agtgaacacg
     2761 gagatcacca aagctccgct gagcgctagt atgatcaagc gctatgatga gcaccaccaa
     2821 gacttgactt tgctgaaggc ccttgtcaga cagcaactgc ctgagaagta caaggaaatt
     2881 ttcttcgatc agtctaaaaa tggctacgcc ggatacattg acggcggagc aagccaggag
     2941 gaattttaca aatttattaa gcccatcttg gaaaaaatgg acggcaccga ggagctgctg
     3001 gtaaagctta acagagaaga tctgttgcgc aaacagcgca ctttcgacaa tggaagcatc
     3061 ccccaccaga ttcacctggg cgaactgcac gctatcctca ggcggcaaga ggatttctac
     3121 ccctttttga aagataacag ggaaaagatt gagaaaatcc tcacatttcg gataccctac
     3181 tatgtaggcc ccctcgcccg gggaaattcc agattcgcgt ggatgactcg caaatcagaa
     3241 gagaccatca ctccctggaa cttcgaggaa gtcgtggata agggggcctc tgcccagtcc
     3301 ttcatcgaaa ggatgactaa ctttgataaa aatctgccta acgaaaaggt gcttcctaaa
     3361 cactctctgc tgtacgagta cttcacagtt tataacgagc tcaccaaggt caaatacgtc
     3421 acagaaggga tgagaaagcc agcattcctg tctggagagc agaagaaagc tatcgtggac
     3481 ctcctcttca agacgaaccg gaaagttacc gtgaaacagc tcaaagaaga ctatttcaaa
     3541 aagattgaat gtttcgactc tgttgaaatc agcggagtgg aggatcgctt caacgcatcc
     3601 ctgggaacgt atcacgatct cctgaaaatc attaaagaca aggacttcct ggacaatgag
     3661 gagaacgagg acattcttga ggacattgtc ctcaccctta cgttgtttga agatagggag
     3721 atgattgaag aacgcttgaa aacttacgct catctcttcg acgacaaagt catgaaacag
     3781 ctcaagaggc gccgatatac aggatggggg cggctgtcaa gaaaactgat caatgggatc
     3841 cgagacaagc agagtggaaa gacaatcctg gattttctta agtccgatgg atttgccaac
     3901 cggaacttca tgcagttgat ccatgatgac tctctcacct ttaaggagga catccagaaa
     3961 gcacaagttt ctggccaggg ggacagtctt cacgagcaca tcgctaatct tgcaggtagc
     4021 ccagctatca aaaagggaat actgcagacc gttaaggtcg tggatgaact cgtcaaagta
     4081 atgggaaggc ataagcccga gaatatcgtt atcgagatgg cccgagagaa ccaaactacc
     4141 cagaagggac agaagaacag tagggaaagg atgaagagga ttgaagaggg tataaaagaa
     4201 ctggggtccc aaatccttaa ggaacaccca gttgaaaaca cccagcttca gaatgagaag
     4261 ctctacctgt actacctgca gaacggcagg gacatgtacg tggatcagga actggacatc
     4321 aatcggctct ccgactacga cgtggatcat atcgtgcccc agtcttttct caaagatgat
     4381 tctattgata ataaagtgtt gacaagatcc gataaaaata gagggaagag tgataacgtc
     4441 ccctcagaag aagttgtcaa gaaaatgaaa aattattggc ggcagctgct gaacgccaaa
     4501 ctgatcacac aacggaagtt cgataatctg actaaggctg aacgaggtgg cctgtctgag
     4561 ttggataaag ccggcttcat caaaaggcag cttgttgaga cacgccagat caccaagcac
     4621 gtggcccaaa ttctcgattc acgcatgaac accaagtacg atgaaaatga caaactgatt
     4681 cgagaggtga aagttattac tctgaagtct aagctggtct cagatttcag aaaggacttt
     4741 cagttttata aggtgagaga gatcaacaat taccaccatg cgcatgatgc ctacctgaat
     4801 gcagtggtag gcactgcact tatcaaaaaa tatcccaagc ttgaatctga atttgtttac
     4861 ggagactata aagtgtacga tgttaggaaa atgatcgcaa agtctgagca ggaaataggc
     4921 aaggccaccg ctaagtactt cttttacagc aatattatga attttttcaa gaccgagatt
     4981 acactggcca atggagagat tcggaagcga ccacttatcg aaacaaacgg agaaacagga
     5041 gaaatcgtgt gggacaaggg tagggatttc gcgacagtcc ggaaggtcct gtccatgccg
     5101 caggtgaaca tcgttaaaaa gaccgaagta cagaccggag gcttctccaa ggaaagtatc
     5161 ctcccgaaaa ggaacagcga caagctgatc gcacgcaaaa aagattggga ccccaagaaa
     5221 tacggcggat tcgattctcc tacagtcgct tacagtgtac tggttgtggc caaagtggag
     5281 aaagggaagt ctaaaaaact caaaagcgtc aaggaactgc tgggcatcac aatcatggag
     5341 cgatcaagct tcgaaaaaaa ccccatcgac tttctcgagg cgaaaggata taaagaggtc
     5401 aaaaaagacc tcatcattaa gcttcccaag tactctctct ttgagcttga aaacggccgg
     5461 aaacgaatgc tcgctagtgc gggcgagctg cagaaaggta acgagctggc actgccctct
     5521 aaatacgtta atttcttgta tctggccagc cactatgaaa agctcaaagg gtctcccgaa
     5581 gataatgagc agaagcagct gttcgtggaa caacacaaac actaccttga tgagatcatc
     5641 gagcaaataa gcgaattctc caaaagagtg atcctcgccg acgctaacct cgataaggtg
     5701 ctttctgctt acaataagca cagggataag cccatcaggg agcaggcaga aaacattatc
     5761 tacttgttta ctctgaccaa cttgggcgcg cctgcagcct tcaagtactt cgacaccacc
     5821 atagacagaa agcggtacac ctctacaaag gaggtcctgg acgccacact gattcatcag
     5881 tcaattacgg ggctctatga aacaagaatc gacctctctc agctcggtgg agacagcagg
     5941 gctgacccca agaagaagag gaaggtgtga tctcttctcg agtcatgtaa ttagttatgt
     6001 cacgcttacg ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac
     6061 aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat
     6121 ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac
     6181 tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc ggccggtacc
     6241 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
     6301 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     6361 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     6421 gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     6481 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     6541 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     6601 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     6661 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6721 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     6781 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     6841 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt
     6901 attttctcct tacgcatctg tgcggtattt cacaccgcat agggtaataa ctgatataat
     6961 taaattgaag ctctaatttg tgagtttagt atacatgcat ttacttataa tacagttttt
     7021 tagttttgct ggccgcatct tctcaaatat gcttcccagc ctgcttttct gtaacgttca
     7081 ccctctacct tagcatccct tccctttgca aatagtcctc ttccaacaat aataatgtca
     7141 gatcctgtag agaccacatc atccacggtt ctatactgtt gacccaatgc gtctcccttg
     7201 tcatctaaac ccacaccggg tgtcataatc aaccaatcgt aaccttcatc tcttccaccc
     7261 atgtctcttt gagcaataaa gccgataaca aaatctttgt cgctcttcgc aatgtcaaca
     7321 gtacccttag tatattctcc agtagatagg gagcccttgc atgacaattc tgctaacatc
     7381 aaaaggcctc taggttcctt tgttacttct tctgccgcct gcttcaaacc gctaacaata
     7441 cctgggccca ccacaccgtg tgcattcgta atgtctgccc attctgctat tctgtataca
     7501 cccgcagagt actgcaattt gactgtatta ccaatgtcag caaattttct gtcttcgaag
     7561 agtaaaaaat tgtacttggc ggataatgcc tttagcggct taactgtgcc ctccatggaa
     7621 aaatcagtca agatatccac atgtgttttt agtaaacaaa ttttgggacc taatgcttca
     7681 actaactcca gtaattcctt ggtggtacga acatccaatg aagcacacaa gtttgtttgc
     7741 ttttcgtgca tgatattaaa tagcttggca gcaacaggac taggatgagt agcagcacgt
     7801 tccttatatg tagctttcga catgatttat cttcgtttcc tgcaggtttt tgttctgtgc
     7861 agttgggtta agaatactgg gcaatttcat gtttcttcaa cactacatat gcgtatatat
     7921 accaatctaa gtctgtgctc cttccttcgt tcttccttct gttcggagat taccgaatca
     7981 aaaaaatttc aaggaaaccg aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaaa
     8041 aggtggtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     8101 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     8161 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     8221 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     8281 taataatggt ttcttagtat gatccaatat caaaggaaat gatagcattg aaggatgaga
     8341 ctaatccaat tgaggagtgg cagcatatag aacagctaaa gggtagtgct gaaggaagca
     8401 tacgataccc cgcatggaat gggataatat cacaggaggt actagactac ctttcatcct
     8461 acataaatag acgcatataa gtacgcattt aagcataaac acgcactatg ccgttcttct
     8521 catgtatata tatatacagg caacacgcag atataggtgc gacgtgaaca gtgagctgta
     8581 tgtgcgcagc tcgcgttgca ttttcggaag cgctcgtttt cggaaacgct ttgaagttcc
     8641 tattccgaag ttcctattct ctagaaagta taggaacttc agagcgcttt tgaaaaccaa
     8701 aagcgctctg aagacgcact ttcaaaaaac caaaaacgca ccggactgta acgagctact
     8761 aaaatattgc gaataccgct tccacaaaca ttgctcaaaa gtatctcttt gctatatatc
     8821 tctgtgctat atccctatat aacctaccca tccacctttc gctccttgaa cttgcatcta
     8881 aactcgacct ctacattttt tatgtttatc tctagtatta ctctttagac aaaaaaattg
     8941 tagtaagaac tattcataga gtgaatcgaa aacaatacga aaatgtaaac atttcctata
     9001 cgtagtatat agagacaaaa tagaagaaac cgttcataat tttctgacca atgaagaatc
     9061 atcaacgcta tcactttctg ttcacaaagt atgcgcaatc cacatcggta tagaatataa
     9121 tcggggatgc ctttatcttg aaaaaatgca cccgcagctt cgctagtaat cagtaaacgc
     9181 gggaagtgga gtcaggcttt ttttatggaa gagaaaatag acaccaaagt agccttcttc
     9241 taaccttaac ggacctacag tgcaaaaagt tatcaagaga ctgcattata gagcgcacaa
     9301 aggagaaaaa aagtaatcta agatgctttg ttagaaaaat agcgctctcg ggatgcattt
     9361 ttgtagaaca aaaaagaagt atagattctt tgttggtaaa atagcgctct cgcgttgcat
     9421 ttctgttctg taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg
     9481 catttttgtt ttacaaaaat gaagcacaga ttcttcgttg gtaaaatagc gctttcgcgt
     9541 tgcatttctg ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt agcgctctcg
     9601 cgttgcattt ttgttctaca aaatgaagca cagatgcttc gttcaggtgg cacttttcgg
     9661 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     9721 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     9781 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     9841 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     9901 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     9961 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
    10021 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
    10081 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
    10141 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
    10201 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
    10261 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
    10321 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
    10381 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
    10441 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
    10501 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
    10561 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
    10621 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
    10681 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
    10741 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
    10801 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
    10861 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
    10921 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
    10981 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gtta
//
