LOCUS       sequence-464598-       10149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast CRISPR plasmid targeting the kanMX and natMX cassettes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10149)
  AUTHORS   Soreanu I, Hendler A, Dahan D, Dovrat D, Aharoni A
  TITLE     Marker-free genetic manipulations in yeast using CRISPR/CAS9 system.
  JOURNAL   Curr Genet. 2018 Oct;64(5):1129-1139. doi: 
            10.1007/s00294-018-0831-y. Epub 2018 Apr 6.
  PUBMED    29626221
REFERENCE   2  (bases 1 to 10149)
  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: 
            "10.1007/s00294-018-0831-y"; journalName: "Curr Genet"; date: 
            "2018-10"; volume: "64"; issue: "5"; pages: "1129-1139"
COMMENT     Sequence Label: pCY5.1kn
FEATURES             Location/Qualifiers
     source          1..10149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        225..300
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        400..615
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             616..1419
                     /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"
     primer_bind     complement(1687..1708)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1796..1900
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1901..1969,1970..2761)
                     /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(2119..2138)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_RNA        3088..3163
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             3959..8062
                     /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
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             3959..8062
                     /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
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8072..8092
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8108..8131
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      8135..8324
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(8381..8969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8461..8480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      8991..9973
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(9954..10001)
                     /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 gtatatgtgt tatgtagtat actctttctt caacaattaa atactctcgg tagccaagtt
       61 ggtttaaggc gcaagactgt aatttatcac tacgaaatct tgagatcggg cgttcgactc
      121 gcccccggga gagatggccg gcatggtccc agcctcctcg ctggcgccgg ctgggcaaca
      181 ccttcgggtg gcgaatggga cttttgtttt gccggggatc gcaggtttta gagctagaaa
      241 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
      301 tttttttatt ttttgtcact attgttatgt aaaatgccac ctctgacagt atggaacgca
      361 aacttctgtc tagtggatat aggtctagag atctaccggt tcaattcatc attttttttt
      421 tattcttttt tttgatttcg gtttccttga aatttttttg attcggtaat ctccgaacag
      481 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
      541 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacctgcagg
      601 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
      661 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
      721 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
      781 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
      841 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaagacag aaaatttgct
      901 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
      961 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
     1021 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     1081 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
     1141 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
     1201 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagacgc attgggtcaa
     1261 cagtatagaa ccgtggatga tgtggtctct acaggatctg acattattat tgttggaaga
     1321 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
     1381 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat
     1441 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attaccctat
     1501 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc
     1561 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     1621 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     1681 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     1741 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcaccgcg gccgccgcgg
     1801 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     1861 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     1921 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     1981 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2041 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2101 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     2161 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     2221 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     2281 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     2341 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     2401 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     2461 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     2521 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     2581 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     2641 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     2701 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     2761 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     2821 taaaaggatc taggtgaaga tcctttttga taatcttaat taagtatatg tgttatgtag
     2881 tatactcttt cttcaacaat taaatactct cggtagccaa gttggtttaa ggcgcaagac
     2941 tgtaatttat cactacgaaa tcttgagatc gggcgttcga ctcgcccccg ggagagatgg
     3001 ccggcatggt cccagcctcc tcgctggcgc cggctgggca acaccttcgg gtggcgaatg
     3061 ggactttgta ccgcaccagt gtcccgggtt ttagagctag aaatagcaag ttaaaataag
     3121 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt attttttgtc
     3181 actattgtta tgtaaaatgc cacctctgac agtatggaac gcaaacttct gtctagtgga
     3241 tactcgagaa cccttaatag tcgaacaaga agcaggcaaa gtttagagca ctgcccctcc
     3301 gcactcaaaa aagaaaaaac taggaggaaa ataaaattct caaccacaca aacacataaa
     3361 cacatacaaa tacaaataca agcttattta cttgacatcg cgcgatcttc cactattcag
     3421 cgccgtccgc cctctctcgt gttttttgtt tacgcgacaa ctatgcgaaa tccggagcaa
     3481 cgggcaaccg tttggggaaa gaccacaccc acgcgcgatc gccatggcaa cgaggtcgca
     3541 cacgccccac acccagacct ccctgcgagc gggcatgggt acaatgtccc cgttgccaca
     3601 gacaccactt cgtagcacag cgcagagcgt agcgtgttgt tgctgctgac aaaagaaaat
     3661 ttttcttagc aaagcaaagg aggggaagca cgggcagata gcaccgtacc atacccttgg
     3721 aaactcgaaa tgaacgaagc aggaaatgag agaatgagag ttttgtaggt atatatagcg
     3781 gtagtgtttg cgcgttacca tcatcttctg gatctatcta ttgttctttt cctcatcact
     3841 ttcccctttt tcgctcttct tcttgtcttt tatttctttc ttttttttaa ttgttccctc
     3901 gattggctat ctaccaaaga atccaaactt aatacacgta tttatttgtc caattaccat
     3961 ggataagaaa tactcaatag gcttagatat cggcacaaat agcgtcggat gggcggtgat
     4021 cactgatgaa tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca
     4081 cagtatcaaa aaaaatctta taggggctct tttatttgac agtggagaga cagcggaagc
     4141 gactcgtctc aaacggacag ctcgtagaag gtatacacgt cggaagaatc gtatttgtta
     4201 tctacaggag attttttcaa atgagatggc gaaagtagat gatagtttct ttcatcgact
     4261 tgaagagtct tttttggtgg aagaagacaa gaagcatgaa cgtcatccta tttttggaaa
     4321 tatagtagat gaagttgctt atcatgagaa atatccaact atctatcatc tgcgaaaaaa
     4381 attggtagat tctactgata aagcggattt gcgcttaatc tatttggcct tagcgcatat
     4441 gattaagttt cgtggtcatt ttttgattga gggagattta aatcctgata atagtgatgt
     4501 ggacaaacta tttatccagt tggtacaaac ctacaatcaa ttatttgaag aaaaccctat
     4561 taacgcaagt ggagtagatg ctaaagcgat tctttctgca cgattgagta aatcaagacg
     4621 attagaaaat ctcattgctc agctccccgg tgagaagaaa aatggcttat ttgggaatct
     4681 cattgctttg tcattgggtt tgacccctaa ttttaaatca aattttgatt tggcagaaga
     4741 tgctaaatta cagctttcaa aagatactta cgatgatgat ttagataatt tattggcgca
     4801 aattggagat caatatgctg atttgttttt ggcagctaag aatttatcag atgctatttt
     4861 actttcagat atcctaagag taaatactga aataactaag gctcccctat cagcttcaat
     4921 gattaaacgc tacgatgaac atcatcaaga cttgactctt ttaaaagctt tagttcgaca
     4981 acaacttcca gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg
     5041 ttatattgat gggggagcta gccaagaaga attttataaa tttatcaaac caattttaga
     5101 aaaaatggat ggtactgagg aattattggt gaaactaaat cgtgaagatt tgctgcgcaa
     5161 gcaacggacc tttgacaacg gctctattcc ccatcaaatt cacttgggtg agctgcatgc
     5221 tattttgaga agacaagaag acttttatcc atttttaaaa gacaatcgtg agaagattga
     5281 aaaaatcttg acttttcgaa ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg
     5341 ttttgcatgg atgactcgga agtctgaaga aacaattacc ccatggaatt ttgaagaagt
     5401 tgtcgataaa ggtgcttcag ctcaatcatt tattgaacgc atgacaaact ttgataaaaa
     5461 tcttccaaat gaaaaagtac taccaaaaca tagtttgctt tatgagtatt ttacggttta
     5521 taacgaattg acaaaggtca aatatgttac tgaaggaatg cgaaaaccag catttctttc
     5581 aggtgaacag aagaaagcca ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt
     5641 taagcaatta aaagaagatt atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc
     5701 aggagttgaa gatagattta atgcttcatt aggtacctac catgatttgc taaaaattat
     5761 taaagataaa gattttttgg ataatgaaga aaatgaagat atcttagagg atattgtttt
     5821 aacattgacc ttatttgaag atagggagat gattgaggaa agacttaaaa catatgctca
     5881 cctctttgat gataaggtga tgaaacagct taaacgtcgc cgttatactg gttggggacg
     5941 tttgtctcga aaattgatta atggtattag ggataagcaa tctggcaaaa caatattaga
     6001 ttttttgaaa tcagatggtt ttgccaatcg caattttatg cagctgatcc atgatgatag
     6061 tttgacattt aaagaagaca ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca
     6121 tgaacatatt gcaaatttag ctggtagccc tgctattaaa aaaggtattt tacagactgt
     6181 aaaagttgtt gatgaattgg tcaaagtaat ggggcggcat aagccagaaa atatcgttat
     6241 tgaaatggca cgtgaaaatc agacaactca aaagggccag aaaaattcgc gagagcgtat
     6301 gaaacgaatc gaagaaggta tcaaagaatt aggaagtcag attcttaaag agcatcctgt
     6361 tgaaaatact caattgcaaa atgaaaagct ctatctctat tatctccaaa atggaagaga
     6421 catgtatgtg gaccaagaat tagatattaa tcgtttaagt gattatgatg tcgatcacat
     6481 tgttccacaa agtttcctta aagacgattc aatagacaat aaggtcttaa cgcgttctga
     6541 taaaaatcgt ggtaaatcgg ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa
     6601 ctattggaga caacttctaa acgccaagtt aatcactcaa cgtaagtttg ataatttaac
     6661 gaaagctgaa cgtggaggtt tgagtgaact tgataaagct ggttttatca aacgccaatt
     6721 ggttgaaact cgccaaatca ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac
     6781 taaatacgat gaaaatgata aacttattcg agaggttaaa gtgattacct taaaatctaa
     6841 attagtttct gacttccgaa aagatttcca attctataaa gtacgtgaga ttaacaatta
     6901 ccatcatgcc catgatgcgt atctaaatgc cgtcgttgga actgctttga ttaagaaata
     6961 tccaaaactt gaatcggagt ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat
     7021 gattgctaag tctgagcaag aaataggcaa agcaaccgca aaatatttct tttactctaa
     7081 tatcatgaac ttcttcaaaa cagaaattac acttgcaaat ggagagattc gcaaacgccc
     7141 tctaatcgaa actaatgggg aaactggaga aattgtctgg gataaagggc gagattttgc
     7201 cacagtgcgc aaagtattgt ccatgcccca agtcaatatt gtcaagaaaa cagaagtaca
     7261 gacaggcgga ttctccaagg agtcaatttt accaaaaaga aattcggaca agcttattgc
     7321 tcgtaaaaaa gactgggatc caaaaaaata tggtggtttt gatagtccaa cggtagctta
     7381 ttcagtccta gtggttgcta aggtggaaaa agggaaatcg aagaagttaa aatccgttaa
     7441 agagttacta gggatcacaa ttatggaaag aagttccttt gaaaaaaatc cgattgactt
     7501 tttagaagct aaaggatata aggaagttaa aaaagactta atcattaaac tacctaaata
     7561 tagtcttttt gagttagaaa acggtcgtaa acggatgctg gctagtgccg gagaattaca
     7621 aaaaggaaat gagctggctc tgccaagcaa atatgtgaat tttttatatt tagctagtca
     7681 ttatgaaaag ttgaagggta gtccagaaga taacgaacaa aaacaattgt ttgtggagca
     7741 gcataagcat tatttagatg agattattga gcaaatcagt gaattttcta agcgtgttat
     7801 tttagcagat gccaatttag ataaagttct tagtgcatat aacaaacata gagacaaacc
     7861 aatacgtgaa caagcagaaa atattattca tttatttacg ttgacgaatc ttggagctcc
     7921 cgctgctttt aaatattttg atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga
     7981 agttttagat gccactctta tccatcaatc catcactggt ctttatgaaa cacgcattga
     8041 tttgagtcag ctaggaggtg acggtggagg gccaaaaaag aaaagaaaag ttgaagatgc
     8101 ttctgctcat catcaccatc accatcacca ttaaatccgc tctaaccgaa aaggaaggag
     8161 ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa
     8221 cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca
     8281 ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaagtgagca aaaggccagc
     8341 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     8401 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     8461 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     8521 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     8581 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     8641 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     8701 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     8761 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     8821 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     8881 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     8941 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct accgaagcat
     9001 ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     9061 atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa
     9121 gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     9181 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca
     9241 acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc
     9301 taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg
     9361 ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt
     9421 ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc
     9481 gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg
     9541 catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga
     9601 acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt
     9661 gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact
     9721 agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt
     9781 ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag
     9841 caatgtttgt ggaagcggta ttcgcaatat tttagtagcc cgttacagtc cggtgcgttt
     9901 ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc tctgaagttc
     9961 ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt ccgaaaacga
    10021 gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac gtcgcaccta
    10081 tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg cgtgtttatg
    10141 cttaaatgc
//
