LOCUS       sequence-193442-       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 ARS1309a sequence 
            CCTGTGGTGACTACGTATCC in yeast chromosome 13..
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-ARS1309a
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..23)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     123..142
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        166..381
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             382..1185
                     /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"
     rep_origin      complement(1316..1771)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1453..1474)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1665..1684
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1897..1919
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1911..1928
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1912..1928
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1938..1957
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1938..1956
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(1975..2222)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(2238..2258)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2271..6374)
                     /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"
     promoter        complement(6384..7088)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     misc_RNA        complement(7174..7249)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(7476..7496)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7476..7494)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7515..7531)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7515..7531)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7528..7550)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7539..7555
                     /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)."
     promoter        complement(join(7563..7569,7570..7587,7588..7593))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7608..7629
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7746..7763)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7917..8505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7997..8016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8676..9467,9468..9536))
                     /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     9299..9318
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9537..9641)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9668..11010
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(10631..10678)
                     /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)."
     primer_bind     11051..11069
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
       61 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
      121 tgcggcatca gagcagattg tactgagagt gcaccatacc accttttcaa ttcatcattt
      181 tttttttatt cttttttttg atttcggttt ccttgaaatt tttttgattc ggtaatctcc
      241 gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt
      301 agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacc
      361 tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat
      421 cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt
      481 gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc
      541 aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca
      601 gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga agacagaaaa
      661 tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca
      721 gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg
      781 aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg
      841 tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag
      901 agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa
      961 ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg agacgcattg
     1021 ggtcaacagt atagaaccgt ggatgatgtg gtctctacag gatctgacat tattattgtt
     1081 ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa
     1141 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa
     1201 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
     1261 ccctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     1321 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     1381 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     1441 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     1501 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     1561 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     1621 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     1681 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     1741 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     1801 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     1861 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     1921 cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc ggccgcaaat
     1981 taaagccttc gagcgtccca aaaccttctc aagcaaggtt ttcagtataa tgttacatgc
     2041 gtacacgcgt ctgtacagaa aaaaaagaaa aatttgaaat ataaataacg ttcttaatac
     2101 taacataact ataaaaaaat aaatagggac ctagacttca ggttgtctaa ctccttcctt
     2161 ttcggttaga gcggatgtgg ggggagggcg tgaacgtaag cgtgacataa ctaattacat
     2221 gactcgagaa gagatcacac cttcctcttc ttcttggggt cagccctgct gtctccaccg
     2281 agctgagaga ggtcgattct tgtttcatag agccccgtaa ttgactgatg aatcagtgtg
     2341 gcgtccagga cctcctttgt agaggtgtac cgctttctgt ctatggtggt gtcgaagtac
     2401 ttgaaggctg caggcgcgcc caagttggtc agagtaaaca agtagataat gttttctgcc
     2461 tgctccctga tgggcttatc cctgtgctta ttgtaagcag aaagcacctt atcgaggtta
     2521 gcgtcggcga ggatcactct tttggagaat tcgcttattt gctcgatgat ctcatcaagg
     2581 tagtgtttgt gttgttccac gaacagctgc ttctgctcat tatcttcggg agaccctttg
     2641 agcttttcat agtggctggc cagatacaag aaattaacgt atttagaggg cagtgccagc
     2701 tcgttacctt tctgcagctc gcccgcacta gcgagcattc gtttccggcc gttttcaagc
     2761 tcaaagagag agtacttggg aagcttaatg atgaggtctt ttttgacctc tttatatcct
     2821 ttcgcctcga gaaagtcgat ggggtttttt tcgaagcttg atcgctccat gattgtgatg
     2881 cccagcagtt ccttgacgct tttgagtttt ttagacttcc ctttctccac tttggccaca
     2941 accagtacac tgtaagcgac tgtaggagaa tcgaatccgc cgtatttctt ggggtcccaa
     3001 tcttttttgc gtgcgatcag cttgtcgctg ttccttttcg ggaggatact ttccttggag
     3061 aagcctccgg tctgtacttc ggtcttttta acgatgttca cctgcggcat ggacaggacc
     3121 ttccggactg tcgcgaaatc cctacccttg tcccacacga tttctcctgt ttctccgttt
     3181 gtttcgataa gtggtcgctt ccgaatctct ccattggcca gtgtaatctc ggtcttgaaa
     3241 aaattcataa tattgctgta aaagaagtac ttagcggtgg ccttgcctat ttcctgctca
     3301 gactttgcga tcattttcct aacatcgtac actttatagt ctccgtaaac aaattcagat
     3361 tcaagcttgg gatatttttt gataagtgca gtgcctacca ctgcattcag gtaggcatca
     3421 tgcgcatggt ggtaattgtt gatctctctc accttataaa actgaaagtc ctttctgaaa
     3481 tctgagacca gcttagactt cagagtaata actttcacct ctcgaatcag tttgtcattt
     3541 tcatcgtact tggtgttcat gcgtgaatcg agaatttggg ccacgtgctt ggtgatctgg
     3601 cgtgtctcaa caagctgcct tttgatgaag ccggctttat ccaactcaga caggccacct
     3661 cgttcagcct tagtcagatt atcgaacttc cgttgtgtga tcagtttggc gttcagcagc
     3721 tgccgccaat aatttttcat tttcttgaca acttcttctg aggggacgtt atcactcttc
     3781 cctctatttt tatcggatct tgtcaacact ttattatcaa tagaatcatc tttgagaaaa
     3841 gactggggca cgatatgatc cacgtcgtag tcggagagcc gattgatgtc cagttcctga
     3901 tccacgtaca tgtccctgcc gttctgcagg tagtacaggt agagcttctc attctgaagc
     3961 tgggtgtttt caactgggtg ttccttaagg atttgggacc ccagttcttt tataccctct
     4021 tcaatcctct tcatcctttc cctactgttc ttctgtccct tctgggtagt ttggttctct
     4081 cgggccatct cgataacgat attctcgggc ttatgccttc ccattacttt gacgagttca
     4141 tccacgacct taacggtctg cagtattccc tttttgatag ctgggctacc tgcaagatta
     4201 gcgatgtgct cgtgaagact gtccccctgg ccagaaactt gtgctttctg gatgtcctcc
     4261 ttaaaggtga gagagtcatc atggatcaac tgcatgaagt tccggttggc aaatccatcg
     4321 gacttaagaa aatccaggat tgtctttcca ctctgcttgt ctcggatccc attgatcagt
     4381 tttcttgaca gccgccccca tcctgtatat cggcgcctct tgagctgttt catgactttg
     4441 tcgtcgaaga gatgagcgta agttttcaag cgttcttcaa tcatctccct atcttcaaac
     4501 aacgtaaggg tgaggacaat gtcctcaaga atgtcctcgt tctcctcatt gtccaggaag
     4561 tccttgtctt taatgatttt caggagatcg tgatacgttc ccagggatgc gttgaagcga
     4621 tcctccactc cgctgatttc aacagagtcg aaacattcaa tctttttgaa atagtcttct
     4681 ttgagctgtt tcacggtaac tttccggttc gtcttgaaga ggaggtccac gatagctttc
     4741 ttctgctctc cagacaggaa tgctggcttt ctcatccctt ctgtgacgta tttgaccttg
     4801 gtgagctcgt tataaactgt gaagtactcg tacagcagag agtgtttagg aagcaccttt
     4861 tcgttaggca gatttttatc aaagttagtc atcctttcga tgaaggactg ggcagaggcc
     4921 cccttatcca cgacttcctc gaagttccag ggagtgatgg tctcttctga tttgcgagtc
     4981 atccacgcga atctggaatt tccccgggcg agggggccta catagtaggg tatccgaaat
     5041 gtgaggattt tctcaatctt ttccctgtta tctttcaaaa aggggtagaa atcctcttgc
     5101 cgcctgagga tagcgtgcag ttcgcccagg tgaatctggt gggggatgct tccattgtcg
     5161 aaagtgcgct gtttgcgcaa cagatcttct ctgttaagct ttaccagcag ctcctcggtg
     5221 ccgtccattt tttccaagat gggcttaata aatttgtaaa attcctcctg gcttgctccg
     5281 ccgtcaatgt atccggcgta gccattttta gactgatcga agaaaatttc cttgtacttc
     5341 tcaggcagtt gctgtctgac aagggccttc agcaaagtca agtcttggtg gtgctcatca
     5401 tagcgcttga tcatactagc gctcagcgga gctttggtga tctccgtgtt cactcgcaga
     5461 atatcactca gcagaatggc gtctgacagg ttctttgccg ccaaaaaaag gtctgcgtac
     5521 tggtcgccga tctgggccag cagattgtcg agatcatcat cgtaggtgtc tttgctcagt
     5581 tgaagcttgg catcttcggc caggtcgaag ttagatttaa agttgggggt cagcccgagt
     5641 gacagggcga taagattacc aaacaggccg ttcttcttct ccccagggag ctgtgcgatg
     5701 aggttttcga gccgccggga tttggacagc ctagcgctca ggattgcttt ggcgtcaact
     5761 ccggatgcgt tgatcgggtt ctcttcgaaa agctgattgt aagtctgaac cagttggata
     5821 aagagtttgt cgacatcgct gttgtctggg ttcaggtccc cctcgatgag gaagtgtccc
     5881 cgaaatttga tcatatgcgc cagcgcgaga tagatcaacc gcaagtcagc cttatcagta
     5941 ctgtctacaa gcttcttcct cagatgatat atggttgggt acttttcatg gtacgccacc
     6001 tcgtccacga tattgccaaa gattgggtgg cgctcgttct ttttatcctc ctccaccaaa
     6061 aaggactcct ccagcctatg gaagaaagag tcatccacct tagccatctc attactaaag
     6121 atctcctgca ggtagcagat ccgattcttt ctgcgggtat atctgcgccg tgctgttctt
     6181 ttgagccgcg tggcttcggc cgtctccccg gagtcgaaca ggagggcgcc aatgaggttc
     6241 ttctttatgc tgtggcgatc ggtattgccc agaactttga attttttgct cggcaccttg
     6301 tactcgtccg taatgacggc ccaaccgacg ctgtttgtgc cgatatcgag cccaatggag
     6361 tacttcttgt ccattttttt ccaagttgat tgtatgcttg gtatagcttg aaatattgtg
     6421 cagaaaaaga aacaaggaag aaagggaacg agaacaatga cgaggaaaca aaagattaat
     6481 aattgcaggt ctatttatac ttgatagcaa gacagcaaac ttttttttat ttcaaattca
     6541 agtaactgga aggaaggccg tataccgttg ctcattagag agtagtgtgc gtgaatgaag
     6601 gaaggaaaaa gtttcgtgtg cttcgagata cccctcatca gctctggaac aacgacatct
     6661 gttggtgctg tctttgtcgt taattttttc ctttagtgtc ttccatcatt tttttgtcat
     6721 tgcggatatg gtgagacaac aacgggggag agagaaaaga aaaaaaaaga aaagaagttg
     6781 catgcgccta ttattacttc aatagatggc aaatggaaaa agggtagtga aacttcgata
     6841 tgatgatggc tatcaagtct agggctacag tattagttcg ttatgtacca ccatcaatga
     6901 ggcagtgtaa ttggtgtagt cttgtttagc ccattatgtc ttgtctggta tctgttctat
     6961 tgtatatctc ccctccgcca cctacatgtt agggagacca acgaaggtat tataggaatc
     7021 ccgatgtatg ggtttggttg ccagaaaaga ggaagtccat attgtacacc cggaaacaac
     7081 aaaaggatgt cgactatcca ctagacagaa gtttgcgttc catactgtca gaggtggcat
     7141 tttacataac aatagtgaca aaaaataaaa aaagcaccga ctcggtgcca ctttttcaag
     7201 ttgataacgg actagcctta ttttaacttg ctatttctag ctctaaaacg gatacgtagt
     7261 caccacagga aagtcccatt cgccacccga aggtgttgcc cagccggcgc cagcgaggag
     7321 gctgggacca tgccggccat ctctcccggg ggcgagtcga acgcccgatc tcaagatttc
     7381 gtagtgataa attacagtct tgcgccttaa accaacttgg ctaccgagag tatttaattg
     7441 ttgaagaaag agtatactag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc
     7501 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     7561 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     7621 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     7681 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     7741 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     7801 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7861 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7921 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7981 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     8041 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     8101 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     8161 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     8221 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     8281 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     8341 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     8401 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     8461 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     8521 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     8581 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     8641 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     8701 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     8761 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     8821 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8881 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8941 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     9001 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     9061 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     9121 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     9181 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     9241 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     9301 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     9361 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     9421 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     9481 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     9541 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     9601 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     9661 ccacctgaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct
     9721 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc
     9781 gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga
     9841 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     9901 agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc
     9961 cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc
    10021 tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct
    10081 actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg
    10141 attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta
    10201 taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat
    10261 tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat
    10321 gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt
    10381 gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag
    10441 ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc
    10501 aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt
    10561 tacagtccgg tgcgtttttg gttttttgaa agtgcgtctt cagagcgctt ttggttttca
    10621 aaagcgctct gaagttccta tactttctag agaataggaa cttcggaata ggaacttcaa
    10681 agcgtttccg aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca tacagctcac
    10741 tgttcacgtc gcacctatat ctgcgtgttg cctgtatata tatatacatg agaagaacgg
    10801 catagtgcgt gtttatgctt aaatgcgtac ttatatgcgt ctatttatgt aggatgaaag
    10861 gtagtctagt acctcctgtg atattatccc attccatgcg gggtatcgta tgcttccttc
    10921 agcactaccc tttagctgtt ctatatgctg ccactcctca attggattag tctcatcctt
    10981 caatgctatc atttcctttg atattggatc atactaagaa accattatta tcatgacatt
    11041 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
    11101 tgaaaa
//
