LOCUS       sequence-193430-       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 ARS607c sequence 
            CTATTTTTGCTTTCTGCACA in yeast chromosome 6..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11106)
  AUTHORS   Reider Apel A, d'Espaux L, Wehrs M, Sachs D, Li RA, Tong GJ, Garber 
            M, Nnadi O, Zhuang W, Hillson NJ, Keasling JD, Mukhopadhyay A
  TITLE     A Cas9-based toolkit to program gene expression in Saccharomyces 
            cerevisiae.
  JOURNAL   Nucleic Acids Res. 2017 Jan 9;45(1):496-508. doi: 
            10.1093/nar/gkw1023. Epub 2016 Nov 28.
  PUBMED    27899650
REFERENCE   2  (bases 1 to 11106)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1093/nar/gkw1023"; journalName: "Nucleic Acids Res"; date: 
            "2017-01-9- 9"; volume: "45"; issue: "1"; pages: "496-508"
COMMENT     Sequence Label: p426_Cas9_gRNA-ARS607c
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..41
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(79..97)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      138..1480
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    470..517
                     /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        1507..1611
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1612..1680,1681..2472)
                     /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(1830..1849)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2643..3231
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3132..3151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3385..3402
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3519..3540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3555..3560,3561..3578,3579..3585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3593..3609
                     /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     3598..3620
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3617..3633
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3617..3633
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3652..3672
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3654..3672
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        3899..3974
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4060..4764
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             4774..8877
                     /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             8890..8910
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8926..9173
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(9191..9210)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9192..9210)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9220..9237)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9220..9236)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9229..9251)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9377..9832
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9464..9483)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9674..9695
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(9963..10766)
                     /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(10767..10982)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(11006..11025)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
       61 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
      121 taatggtttc ttagtatgat ccaatatcaa aggaaatgat agcattgaag gatgagacta
      181 atccaattga ggagtggcag catatagaac agctaaaggg tagtgctgaa ggaagcatac
      241 gataccccgc atggaatggg ataatatcac aggaggtact agactacctt tcatcctaca
      301 taaatagacg catataagta cgcatttaag cataaacacg cactatgccg ttcttctcat
      361 gtatatatat atacaggcaa cacgcagata taggtgcgac gtgaacagtg agctgtatgt
      421 gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg aaacgctttg aagttcctat
      481 tccgaagttc ctattctcta gaaagtatag gaacttcaga gcgcttttga aaaccaaaag
      541 cgctctgaag acgcactttc aaaaaaccaa aaacgcaccg gactgtaacg agctactaaa
      601 atattgcgaa taccgcttcc acaaacattg ctcaaaagta tctctttgct atatatctct
      661 gtgctatatc cctatataac ctacccatcc acctttcgct ccttgaactt gcatctaaac
      721 tcgacctcta cattttttat gtttatctct agtattactc tttagacaaa aaaattgtag
      781 taagaactat tcatagagtg aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt
      841 agtatataga gacaaaatag aagaaaccgt tcataatttt ctgaccaatg aagaatcatc
      901 aacgctatca ctttctgttc acaaagtatg cgcaatccac atcggtatag aatataatcg
      961 gggatgcctt tatcttgaaa aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg
     1021 aagtggagtc aggctttttt tatggaagag aaaatagaca ccaaagtagc cttcttctaa
     1081 ccttaacgga cctacagtgc aaaaagttat caagagactg cattatagag cgcacaaagg
     1141 agaaaaaaag taatctaaga tgctttgtta gaaaaatagc gctctcggga tgcatttttg
     1201 tagaacaaaa aagaagtata gattctttgt tggtaaaata gcgctctcgc gttgcatttc
     1261 tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat
     1321 ttttgtttta caaaaatgaa gcacagattc ttcgttggta aaatagcgct ttcgcgttgc
     1381 atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt
     1441 tgcatttttg ttctacaaaa tgaagcacag atgcttcgtt caggtggcac ttttcgggga
     1501 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1561 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1621 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1681 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1741 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1801 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     1861 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     1921 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     1981 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2041 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2101 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2161 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2221 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2281 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2341 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2401 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2461 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2521 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2581 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2641 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2701 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2761 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2821 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2881 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2941 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3001 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3061 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3241 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3301 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3361 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     3421 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     3481 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     3541 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     3601 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
     3661 cctcactaaa gggaacaaaa gctggagcta gtatactctt tcttcaacaa ttaaatactc
     3721 tcggtagcca agttggttta aggcgcaaga ctgtaattta tcactacgaa atcttgagat
     3781 cgggcgttcg actcgccccc gggagagatg gccggcatgg tcccagcctc ctcgctggcg
     3841 ccggctgggc aacaccttcg ggtggcgaat gggactttct atttttgctt tctgcacagt
     3901 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
     3961 caccgagtcg gtgctttttt tattttttgt cactattgtt atgtaaaatg ccacctctga
     4021 cagtatggaa cgcaaacttc tgtctagtgg atagtcgaca tccttttgtt gtttccgggt
     4081 gtacaatatg gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata
     4141 ccttcgttgg tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac
     4201 cagacaagac ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg
     4261 tacataacga actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca
     4321 ctaccctttt tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct
     4381 ttttttttct tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat
     4441 gatggaagac actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt
     4501 ccagagctga tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca
     4561 cactactctc taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata
     4621 aaaaaaagtt tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt
     4681 ttcctcgtca ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa
     4741 gctataccaa gcatacaatc aacttggaaa aaaatggaca agaagtactc cattgggctc
     4801 gatatcggca caaacagcgt cggttgggcc gtcattacgg acgagtacaa ggtgccgagc
     4861 aaaaaattca aagttctggg caataccgat cgccacagca taaagaagaa cctcattggc
     4921 gccctcctgt tcgactccgg ggagacggcc gaagccacgc ggctcaaaag aacagcacgg
     4981 cgcagatata cccgcagaaa gaatcggatc tgctacctgc aggagatctt tagtaatgag
     5041 atggctaagg tggatgactc tttcttccat aggctggagg agtccttttt ggtggaggag
     5101 gataaaaaga acgagcgcca cccaatcttt ggcaatatcg tggacgaggt ggcgtaccat
     5161 gaaaagtacc caaccatata tcatctgagg aagaagcttg tagacagtac tgataaggct
     5221 gacttgcggt tgatctatct cgcgctggcg catatgatca aatttcgggg acacttcctc
     5281 atcgaggggg acctgaaccc agacaacagc gatgtcgaca aactctttat ccaactggtt
     5341 cagacttaca atcagctttt cgaagagaac ccgatcaacg catccggagt tgacgccaaa
     5401 gcaatcctga gcgctaggct gtccaaatcc cggcggctcg aaaacctcat cgcacagctc
     5461 cctggggaga agaagaacgg cctgtttggt aatcttatcg ccctgtcact cgggctgacc
     5521 cccaacttta aatctaactt cgacctggcc gaagatgcca agcttcaact gagcaaagac
     5581 acctacgatg atgatctcga caatctgctg gcccagatcg gcgaccagta cgcagacctt
     5641 tttttggcgg caaagaacct gtcagacgcc attctgctga gtgatattct gcgagtgaac
     5701 acggagatca ccaaagctcc gctgagcgct agtatgatca agcgctatga tgagcaccac
     5761 caagacttga ctttgctgaa ggcccttgtc agacagcaac tgcctgagaa gtacaaggaa
     5821 attttcttcg atcagtctaa aaatggctac gccggataca ttgacggcgg agcaagccag
     5881 gaggaatttt acaaatttat taagcccatc ttggaaaaaa tggacggcac cgaggagctg
     5941 ctggtaaagc ttaacagaga agatctgttg cgcaaacagc gcactttcga caatggaagc
     6001 atcccccacc agattcacct gggcgaactg cacgctatcc tcaggcggca agaggatttc
     6061 tacccctttt tgaaagataa cagggaaaag attgagaaaa tcctcacatt tcggataccc
     6121 tactatgtag gccccctcgc ccggggaaat tccagattcg cgtggatgac tcgcaaatca
     6181 gaagagacca tcactccctg gaacttcgag gaagtcgtgg ataagggggc ctctgcccag
     6241 tccttcatcg aaaggatgac taactttgat aaaaatctgc ctaacgaaaa ggtgcttcct
     6301 aaacactctc tgctgtacga gtacttcaca gtttataacg agctcaccaa ggtcaaatac
     6361 gtcacagaag ggatgagaaa gccagcattc ctgtctggag agcagaagaa agctatcgtg
     6421 gacctcctct tcaagacgaa ccggaaagtt accgtgaaac agctcaaaga agactatttc
     6481 aaaaagattg aatgtttcga ctctgttgaa atcagcggag tggaggatcg cttcaacgca
     6541 tccctgggaa cgtatcacga tctcctgaaa atcattaaag acaaggactt cctggacaat
     6601 gaggagaacg aggacattct tgaggacatt gtcctcaccc ttacgttgtt tgaagatagg
     6661 gagatgattg aagaacgctt gaaaacttac gctcatctct tcgacgacaa agtcatgaaa
     6721 cagctcaaga ggcgccgata tacaggatgg gggcggctgt caagaaaact gatcaatggg
     6781 atccgagaca agcagagtgg aaagacaatc ctggattttc ttaagtccga tggatttgcc
     6841 aaccggaact tcatgcagtt gatccatgat gactctctca cctttaagga ggacatccag
     6901 aaagcacaag tttctggcca gggggacagt cttcacgagc acatcgctaa tcttgcaggt
     6961 agcccagcta tcaaaaaggg aatactgcag accgttaagg tcgtggatga actcgtcaaa
     7021 gtaatgggaa ggcataagcc cgagaatatc gttatcgaga tggcccgaga gaaccaaact
     7081 acccagaagg gacagaagaa cagtagggaa aggatgaaga ggattgaaga gggtataaaa
     7141 gaactggggt cccaaatcct taaggaacac ccagttgaaa acacccagct tcagaatgag
     7201 aagctctacc tgtactacct gcagaacggc agggacatgt acgtggatca ggaactggac
     7261 atcaatcggc tctccgacta cgacgtggat catatcgtgc cccagtcttt tctcaaagat
     7321 gattctattg ataataaagt gttgacaaga tccgataaaa atagagggaa gagtgataac
     7381 gtcccctcag aagaagttgt caagaaaatg aaaaattatt ggcggcagct gctgaacgcc
     7441 aaactgatca cacaacggaa gttcgataat ctgactaagg ctgaacgagg tggcctgtct
     7501 gagttggata aagccggctt catcaaaagg cagcttgttg agacacgcca gatcaccaag
     7561 cacgtggccc aaattctcga ttcacgcatg aacaccaagt acgatgaaaa tgacaaactg
     7621 attcgagagg tgaaagttat tactctgaag tctaagctgg tctcagattt cagaaaggac
     7681 tttcagtttt ataaggtgag agagatcaac aattaccacc atgcgcatga tgcctacctg
     7741 aatgcagtgg taggcactgc acttatcaaa aaatatccca agcttgaatc tgaatttgtt
     7801 tacggagact ataaagtgta cgatgttagg aaaatgatcg caaagtctga gcaggaaata
     7861 ggcaaggcca ccgctaagta cttcttttac agcaatatta tgaatttttt caagaccgag
     7921 attacactgg ccaatggaga gattcggaag cgaccactta tcgaaacaaa cggagaaaca
     7981 ggagaaatcg tgtgggacaa gggtagggat ttcgcgacag tccggaaggt cctgtccatg
     8041 ccgcaggtga acatcgttaa aaagaccgaa gtacagaccg gaggcttctc caaggaaagt
     8101 atcctcccga aaaggaacag cgacaagctg atcgcacgca aaaaagattg ggaccccaag
     8161 aaatacggcg gattcgattc tcctacagtc gcttacagtg tactggttgt ggccaaagtg
     8221 gagaaaggga agtctaaaaa actcaaaagc gtcaaggaac tgctgggcat cacaatcatg
     8281 gagcgatcaa gcttcgaaaa aaaccccatc gactttctcg aggcgaaagg atataaagag
     8341 gtcaaaaaag acctcatcat taagcttccc aagtactctc tctttgagct tgaaaacggc
     8401 cggaaacgaa tgctcgctag tgcgggcgag ctgcagaaag gtaacgagct ggcactgccc
     8461 tctaaatacg ttaatttctt gtatctggcc agccactatg aaaagctcaa agggtctccc
     8521 gaagataatg agcagaagca gctgttcgtg gaacaacaca aacactacct tgatgagatc
     8581 atcgagcaaa taagcgaatt ctccaaaaga gtgatcctcg ccgacgctaa cctcgataag
     8641 gtgctttctg cttacaataa gcacagggat aagcccatca gggagcaggc agaaaacatt
     8701 atctacttgt ttactctgac caacttgggc gcgcctgcag ccttcaagta cttcgacacc
     8761 accatagaca gaaagcggta cacctctaca aaggaggtcc tggacgccac actgattcat
     8821 cagtcaatta cggggctcta tgaaacaaga atcgacctct ctcagctcgg tggagacagc
     8881 agggctgacc ccaagaagaa gaggaaggtg tgatctcttc tcgagtcatg taattagtta
     8941 tgtcacgctt acgttcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta
     9001 gacaacctga agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt
     9061 tatttatatt tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta
     9121 tactgaaaac cttgcttgag aaggttttgg gacgctcgaa ggctttaatt tgcggccggt
     9181 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     9241 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     9301 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     9361 cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     9421 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     9481 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     9541 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     9601 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     9661 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     9721 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     9781 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc
     9841 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catagggtaa taactgatat
     9901 aattaaattg aagctctaat ttgtgagttt agtatacatg catttactta taatacagtt
     9961 ttttagtttt gctggccgca tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt
    10021 tcaccctcta ccttagcatc ccttcccttt gcaaatagtc ctcttccaac aataataatg
    10081 tcagatcctg tagagaccac atcatccacg gttctatact gttgacccaa tgcgtctccc
    10141 ttgtcatcta aacccacacc gggtgtcata atcaaccaat cgtaaccttc atctcttcca
    10201 cccatgtctc tttgagcaat aaagccgata acaaaatctt tgtcgctctt cgcaatgtca
    10261 acagtaccct tagtatattc tccagtagat agggagccct tgcatgacaa ttctgctaac
    10321 atcaaaaggc ctctaggttc ctttgttact tcttctgccg cctgcttcaa accgctaaca
    10381 atacctgggc ccaccacacc gtgtgcattc gtaatgtctg cccattctgc tattctgtat
    10441 acacccgcag agtactgcaa tttgactgta ttaccaatgt cagcaaattt tctgtcttcg
    10501 aagagtaaaa aattgtactt ggcggataat gcctttagcg gcttaactgt gccctccatg
    10561 gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac aaattttggg acctaatgct
    10621 tcaactaact ccagtaattc cttggtggta cgaacatcca atgaagcaca caagtttgtt
    10681 tgcttttcgt gcatgatatt aaatagcttg gcagcaacag gactaggatg agtagcagca
    10741 cgttccttat atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg
    10801 tgcagttggg ttaagaatac tgggcaattt catgtttctt caacactaca tatgcgtata
    10861 tataccaatc taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa
    10921 tcaaaaaaat ttcaaggaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg
    10981 aaaaggtggt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
    11041 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
    11101 cttaca
//
