LOCUS       sequence-193445-       11105 bp    DNA     circular SYN 15-APR-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 HIS3b sequence 
            AATATAGAGTGTACTAGAGG in yeast chromosome 15..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11105)
  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 11105)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-HIS3b
FEATURES             Location/Qualifiers
     source          1..11105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        838..942
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(943..1011,1012..1803)
                     /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(1161..1180)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1974..2562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2463..2482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2716..2733
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2850..2871
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2886..2891,2892..2909,2910..2916)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2924..2940
                     /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     2929..2951
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2948..2964
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2948..2964
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2983..3003
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2985..3003
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        3229..3304
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3390..4094
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             4104..8207
                     /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             8220..8240
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8256..8503
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(8521..8540)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8522..8540)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8550..8567)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8550..8566)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8559..8581)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8707..9162
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8794..8813)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9004..9025
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(9293..10096)
                     /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(10097..10312)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(10336..10355)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10455..10477
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(10515..10533)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      join(10574..11105,1..811)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    10906..10953
                     /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)."
ORIGIN
        1 ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct
       61 acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta
      121 ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag
      181 agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc
      241 actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct
      301 ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt
      361 caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg
      421 acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa
      481 gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa
      541 aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt ctgttctgta
      601 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt
      661 acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg catttctgtt
      721 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
      781 gttctacaaa atgaagcaca gatgcttcgt tcaggtggca cttttcgggg aaatgtgcgc
      841 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
      901 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
      961 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1021 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1081 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1141 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1201 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1261 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1321 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1381 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     1441 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     1501 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1561 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1621 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     1681 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     1741 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1801 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     1861 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     1921 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1981 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2041 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2101 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2161 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2221 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2281 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2341 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2401 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2461 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2521 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2581 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2641 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2701 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     2761 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     2821 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     2881 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     2941 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     3001 agggaacaaa agctggagct agtatactct ttcttcaaca attaaatact ctcggtagcc
     3061 aagttggttt aaggcgcaag actgtaattt atcactacga aatcttgaga tcgggcgttc
     3121 gactcgcccc cgggagagat ggccggcatg gtcccagcct cctcgctggc gccggctggg
     3181 caacaccttc gggtggcgaa tgggacttta atatagagtg tactagaggg tttagagcta
     3241 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     3301 gtgctttttt tattttttgt cactattgtt atgtaaaatg ccacctctga cagtatggaa
     3361 cgcaaacttc tgtctagtgg atagtcgaca tccttttgtt gtttccgggt gtacaatatg
     3421 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
     3481 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
     3541 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
     3601 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
     3661 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
     3721 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
     3781 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
     3841 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
     3901 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
     3961 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
     4021 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
     4081 gcatacaatc aacttggaaa aaaatggaca agaagtactc cattgggctc gatatcggca
     4141 caaacagcgt cggttgggcc gtcattacgg acgagtacaa ggtgccgagc aaaaaattca
     4201 aagttctggg caataccgat cgccacagca taaagaagaa cctcattggc gccctcctgt
     4261 tcgactccgg ggagacggcc gaagccacgc ggctcaaaag aacagcacgg cgcagatata
     4321 cccgcagaaa gaatcggatc tgctacctgc aggagatctt tagtaatgag atggctaagg
     4381 tggatgactc tttcttccat aggctggagg agtccttttt ggtggaggag gataaaaaga
     4441 acgagcgcca cccaatcttt ggcaatatcg tggacgaggt ggcgtaccat gaaaagtacc
     4501 caaccatata tcatctgagg aagaagcttg tagacagtac tgataaggct gacttgcggt
     4561 tgatctatct cgcgctggcg catatgatca aatttcgggg acacttcctc atcgaggggg
     4621 acctgaaccc agacaacagc gatgtcgaca aactctttat ccaactggtt cagacttaca
     4681 atcagctttt cgaagagaac ccgatcaacg catccggagt tgacgccaaa gcaatcctga
     4741 gcgctaggct gtccaaatcc cggcggctcg aaaacctcat cgcacagctc cctggggaga
     4801 agaagaacgg cctgtttggt aatcttatcg ccctgtcact cgggctgacc cccaacttta
     4861 aatctaactt cgacctggcc gaagatgcca agcttcaact gagcaaagac acctacgatg
     4921 atgatctcga caatctgctg gcccagatcg gcgaccagta cgcagacctt tttttggcgg
     4981 caaagaacct gtcagacgcc attctgctga gtgatattct gcgagtgaac acggagatca
     5041 ccaaagctcc gctgagcgct agtatgatca agcgctatga tgagcaccac caagacttga
     5101 ctttgctgaa ggcccttgtc agacagcaac tgcctgagaa gtacaaggaa attttcttcg
     5161 atcagtctaa aaatggctac gccggataca ttgacggcgg agcaagccag gaggaatttt
     5221 acaaatttat taagcccatc ttggaaaaaa tggacggcac cgaggagctg ctggtaaagc
     5281 ttaacagaga agatctgttg cgcaaacagc gcactttcga caatggaagc atcccccacc
     5341 agattcacct gggcgaactg cacgctatcc tcaggcggca agaggatttc tacccctttt
     5401 tgaaagataa cagggaaaag attgagaaaa tcctcacatt tcggataccc tactatgtag
     5461 gccccctcgc ccggggaaat tccagattcg cgtggatgac tcgcaaatca gaagagacca
     5521 tcactccctg gaacttcgag gaagtcgtgg ataagggggc ctctgcccag tccttcatcg
     5581 aaaggatgac taactttgat aaaaatctgc ctaacgaaaa ggtgcttcct aaacactctc
     5641 tgctgtacga gtacttcaca gtttataacg agctcaccaa ggtcaaatac gtcacagaag
     5701 ggatgagaaa gccagcattc ctgtctggag agcagaagaa agctatcgtg gacctcctct
     5761 tcaagacgaa ccggaaagtt accgtgaaac agctcaaaga agactatttc aaaaagattg
     5821 aatgtttcga ctctgttgaa atcagcggag tggaggatcg cttcaacgca tccctgggaa
     5881 cgtatcacga tctcctgaaa atcattaaag acaaggactt cctggacaat gaggagaacg
     5941 aggacattct tgaggacatt gtcctcaccc ttacgttgtt tgaagatagg gagatgattg
     6001 aagaacgctt gaaaacttac gctcatctct tcgacgacaa agtcatgaaa cagctcaaga
     6061 ggcgccgata tacaggatgg gggcggctgt caagaaaact gatcaatggg atccgagaca
     6121 agcagagtgg aaagacaatc ctggattttc ttaagtccga tggatttgcc aaccggaact
     6181 tcatgcagtt gatccatgat gactctctca cctttaagga ggacatccag aaagcacaag
     6241 tttctggcca gggggacagt cttcacgagc acatcgctaa tcttgcaggt agcccagcta
     6301 tcaaaaaggg aatactgcag accgttaagg tcgtggatga actcgtcaaa gtaatgggaa
     6361 ggcataagcc cgagaatatc gttatcgaga tggcccgaga gaaccaaact acccagaagg
     6421 gacagaagaa cagtagggaa aggatgaaga ggattgaaga gggtataaaa gaactggggt
     6481 cccaaatcct taaggaacac ccagttgaaa acacccagct tcagaatgag aagctctacc
     6541 tgtactacct gcagaacggc agggacatgt acgtggatca ggaactggac atcaatcggc
     6601 tctccgacta cgacgtggat catatcgtgc cccagtcttt tctcaaagat gattctattg
     6661 ataataaagt gttgacaaga tccgataaaa atagagggaa gagtgataac gtcccctcag
     6721 aagaagttgt caagaaaatg aaaaattatt ggcggcagct gctgaacgcc aaactgatca
     6781 cacaacggaa gttcgataat ctgactaagg ctgaacgagg tggcctgtct gagttggata
     6841 aagccggctt catcaaaagg cagcttgttg agacacgcca gatcaccaag cacgtggccc
     6901 aaattctcga ttcacgcatg aacaccaagt acgatgaaaa tgacaaactg attcgagagg
     6961 tgaaagttat tactctgaag tctaagctgg tctcagattt cagaaaggac tttcagtttt
     7021 ataaggtgag agagatcaac aattaccacc atgcgcatga tgcctacctg aatgcagtgg
     7081 taggcactgc acttatcaaa aaatatccca agcttgaatc tgaatttgtt tacggagact
     7141 ataaagtgta cgatgttagg aaaatgatcg caaagtctga gcaggaaata ggcaaggcca
     7201 ccgctaagta cttcttttac agcaatatta tgaatttttt caagaccgag attacactgg
     7261 ccaatggaga gattcggaag cgaccactta tcgaaacaaa cggagaaaca ggagaaatcg
     7321 tgtgggacaa gggtagggat ttcgcgacag tccggaaggt cctgtccatg ccgcaggtga
     7381 acatcgttaa aaagaccgaa gtacagaccg gaggcttctc caaggaaagt atcctcccga
     7441 aaaggaacag cgacaagctg atcgcacgca aaaaagattg ggaccccaag aaatacggcg
     7501 gattcgattc tcctacagtc gcttacagtg tactggttgt ggccaaagtg gagaaaggga
     7561 agtctaaaaa actcaaaagc gtcaaggaac tgctgggcat cacaatcatg gagcgatcaa
     7621 gcttcgaaaa aaaccccatc gactttctcg aggcgaaagg atataaagag gtcaaaaaag
     7681 acctcatcat taagcttccc aagtactctc tctttgagct tgaaaacggc cggaaacgaa
     7741 tgctcgctag tgcgggcgag ctgcagaaag gtaacgagct ggcactgccc tctaaatacg
     7801 ttaatttctt gtatctggcc agccactatg aaaagctcaa agggtctccc gaagataatg
     7861 agcagaagca gctgttcgtg gaacaacaca aacactacct tgatgagatc atcgagcaaa
     7921 taagcgaatt ctccaaaaga gtgatcctcg ccgacgctaa cctcgataag gtgctttctg
     7981 cttacaataa gcacagggat aagcccatca gggagcaggc agaaaacatt atctacttgt
     8041 ttactctgac caacttgggc gcgcctgcag ccttcaagta cttcgacacc accatagaca
     8101 gaaagcggta cacctctaca aaggaggtcc tggacgccac actgattcat cagtcaatta
     8161 cggggctcta tgaaacaaga atcgacctct ctcagctcgg tggagacagc agggctgacc
     8221 ccaagaagaa gaggaaggtg tgatctcttc tcgagtcatg taattagtta tgtcacgctt
     8281 acgttcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     8341 agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt
     8401 tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac
     8461 cttgcttgag aaggttttgg gacgctcgaa ggctttaatt tgcggccggt acccaattcg
     8521 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     8581 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     8641 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     8701 gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     8761 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     8821 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     8881 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     8941 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     9001 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     9061 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     9121 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     9181 ccttacgcat ctgtgcggta tttcacaccg catagggtaa taactgatat aattaaattg
     9241 aagctctaat ttgtgagttt agtatacatg catttactta taatacagtt ttttagtttt
     9301 gctggccgca tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt tcaccctcta
     9361 ccttagcatc ccttcccttt gcaaatagtc ctcttccaac aataataatg tcagatcctg
     9421 tagagaccac atcatccacg gttctatact gttgacccaa tgcgtctccc ttgtcatcta
     9481 aacccacacc gggtgtcata atcaaccaat cgtaaccttc atctcttcca cccatgtctc
     9541 tttgagcaat aaagccgata acaaaatctt tgtcgctctt cgcaatgtca acagtaccct
     9601 tagtatattc tccagtagat agggagccct tgcatgacaa ttctgctaac atcaaaaggc
     9661 ctctaggttc ctttgttact tcttctgccg cctgcttcaa accgctaaca atacctgggc
     9721 ccaccacacc gtgtgcattc gtaatgtctg cccattctgc tattctgtat acacccgcag
     9781 agtactgcaa tttgactgta ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa
     9841 aattgtactt ggcggataat gcctttagcg gcttaactgt gccctccatg gaaaaatcag
     9901 tcaagatatc cacatgtgtt tttagtaaac aaattttggg acctaatgct tcaactaact
     9961 ccagtaattc cttggtggta cgaacatcca atgaagcaca caagtttgtt tgcttttcgt
    10021 gcatgatatt aaatagcttg gcagcaacag gactaggatg agtagcagca cgttccttat
    10081 atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg tgcagttggg
    10141 ttaagaatac tgggcaattt catgtttctt caacactaca tatgcgtata tataccaatc
    10201 taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat
    10261 ttcaaggaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg aaaaggtggt
    10321 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
    10381 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
    10441 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
    10501 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
    10561 ggtttcttag tatgatccaa tatcaaagga aatgatagca ttgaaggatg agactaatcc
    10621 aattgaggag tggcagcata tagaacagct aaagggtagt gctgaaggaa gcatacgata
    10681 ccccgcatgg aatgggataa tatcacagga ggtactagac tacctttcat cctacataaa
    10741 tagacgcata taagtacgca tttaagcata aacacgcact atgccgttct tctcatgtat
    10801 atatatatac aggcaacacg cagatatagg tgcgacgtga acagtgagct gtatgtgcgc
    10861 agctcgcgtt gcattttcgg aagcgctcgt tttcggaaac gctttgaagt tcctattccg
    10921 aagttcctat tctctagaaa gtataggaac ttcagagcgc ttttgaaaac caaaagcgct
    10981 ctgaagacgc actttcaaaa aaccaaaaac gcaccggact gtaacgagct actaaaatat
    11041 tgcgaatacc gcttccacaa acattgctca aaagtatctc tttgctatat atctctgtgc
    11101 tatat
//
