LOCUS       sequence-477739-       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 ARS511b sequence 
            CAGTGTATGCCAGTCAGCCA in yeast chromosome 5..
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-ARS511b
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    520..541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(556..561,562..579,580..586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    594..610
                     /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     599..621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     618..634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     618..634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     653..673
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        655..673
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        900..975
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1061..1765
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1775..5878
                     /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             5891..5911
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5927..6174
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6192..6211)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6193..6211)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6221..6238)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6221..6237)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6230..6252)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6378..6833
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6465..6484)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6675..6696
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6964..7767)
                     /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(7768..7983)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(8007..8026)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8126..8148
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8186..8204)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      8245..9587
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    8577..8624
                     /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        9614..9718
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9719..9787,9788..10579)
                     /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(9937..9956)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(10750..11106,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      541 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      601 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
      661 ccctcactaa agggaacaaa agctggagct agtatactct ttcttcaaca attaaatact
      721 ctcggtagcc aagttggttt aaggcgcaag actgtaattt atcactacga aatcttgaga
      781 tcgggcgttc gactcgcccc cgggagagat ggccggcatg gtcccagcct cctcgctggc
      841 gccggctggg caacaccttc gggtggcgaa tgggactttc agtgtatgcc agtcagccag
      901 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
      961 gcaccgagtc ggtgcttttt ttattttttg tcactattgt tatgtaaaat gccacctctg
     1021 acagtatgga acgcaaactt ctgtctagtg gatagtcgac atccttttgt tgtttccggg
     1081 tgtacaatat ggacttcctc ttttctggca accaaaccca tacatcggga ttcctataat
     1141 accttcgttg gtctccctaa catgtaggtg gcggagggga gatatacaat agaacagata
     1201 ccagacaaga cataatgggc taaacaagac tacaccaatt acactgcctc attgatggtg
     1261 gtacataacg aactaatact gtagccctag acttgatagc catcatcata tcgaagtttc
     1321 actacccttt ttccatttgc catctattga agtaataata ggcgcatgca acttcttttc
     1381 tttttttttc ttttctctct cccccgttgt tgtctcacca tatccgcaat gacaaaaaaa
     1441 tgatggaaga cactaaagga aaaaattaac gacaaagaca gcaccaacag atgtcgttgt
     1501 tccagagctg atgaggggta tctcgaagca cacgaaactt tttccttcct tcattcacgc
     1561 acactactct ctaatgagca acggtatacg gccttccttc cagttacttg aatttgaaat
     1621 aaaaaaaagt ttgctgtctt gctatcaagt ataaatagac ctgcaattat taatcttttg
     1681 tttcctcgtc attgttctcg ttccctttct tccttgtttc tttttctgca caatatttca
     1741 agctatacca agcatacaat caacttggaa aaaaatggac aagaagtact ccattgggct
     1801 cgatatcggc acaaacagcg tcggttgggc cgtcattacg gacgagtaca aggtgccgag
     1861 caaaaaattc aaagttctgg gcaataccga tcgccacagc ataaagaaga acctcattgg
     1921 cgccctcctg ttcgactccg gggagacggc cgaagccacg cggctcaaaa gaacagcacg
     1981 gcgcagatat acccgcagaa agaatcggat ctgctacctg caggagatct ttagtaatga
     2041 gatggctaag gtggatgact ctttcttcca taggctggag gagtcctttt tggtggagga
     2101 ggataaaaag aacgagcgcc acccaatctt tggcaatatc gtggacgagg tggcgtacca
     2161 tgaaaagtac ccaaccatat atcatctgag gaagaagctt gtagacagta ctgataaggc
     2221 tgacttgcgg ttgatctatc tcgcgctggc gcatatgatc aaatttcggg gacacttcct
     2281 catcgagggg gacctgaacc cagacaacag cgatgtcgac aaactcttta tccaactggt
     2341 tcagacttac aatcagcttt tcgaagagaa cccgatcaac gcatccggag ttgacgccaa
     2401 agcaatcctg agcgctaggc tgtccaaatc ccggcggctc gaaaacctca tcgcacagct
     2461 ccctggggag aagaagaacg gcctgtttgg taatcttatc gccctgtcac tcgggctgac
     2521 ccccaacttt aaatctaact tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga
     2581 cacctacgat gatgatctcg acaatctgct ggcccagatc ggcgaccagt acgcagacct
     2641 ttttttggcg gcaaagaacc tgtcagacgc cattctgctg agtgatattc tgcgagtgaa
     2701 cacggagatc accaaagctc cgctgagcgc tagtatgatc aagcgctatg atgagcacca
     2761 ccaagacttg actttgctga aggcccttgt cagacagcaa ctgcctgaga agtacaagga
     2821 aattttcttc gatcagtcta aaaatggcta cgccggatac attgacggcg gagcaagcca
     2881 ggaggaattt tacaaattta ttaagcccat cttggaaaaa atggacggca ccgaggagct
     2941 gctggtaaag cttaacagag aagatctgtt gcgcaaacag cgcactttcg acaatggaag
     3001 catcccccac cagattcacc tgggcgaact gcacgctatc ctcaggcggc aagaggattt
     3061 ctaccccttt ttgaaagata acagggaaaa gattgagaaa atcctcacat ttcggatacc
     3121 ctactatgta ggccccctcg cccggggaaa ttccagattc gcgtggatga ctcgcaaatc
     3181 agaagagacc atcactccct ggaacttcga ggaagtcgtg gataaggggg cctctgccca
     3241 gtccttcatc gaaaggatga ctaactttga taaaaatctg cctaacgaaa aggtgcttcc
     3301 taaacactct ctgctgtacg agtacttcac agtttataac gagctcacca aggtcaaata
     3361 cgtcacagaa gggatgagaa agccagcatt cctgtctgga gagcagaaga aagctatcgt
     3421 ggacctcctc ttcaagacga accggaaagt taccgtgaaa cagctcaaag aagactattt
     3481 caaaaagatt gaatgtttcg actctgttga aatcagcgga gtggaggatc gcttcaacgc
     3541 atccctggga acgtatcacg atctcctgaa aatcattaaa gacaaggact tcctggacaa
     3601 tgaggagaac gaggacattc ttgaggacat tgtcctcacc cttacgttgt ttgaagatag
     3661 ggagatgatt gaagaacgct tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa
     3721 acagctcaag aggcgccgat atacaggatg ggggcggctg tcaagaaaac tgatcaatgg
     3781 gatccgagac aagcagagtg gaaagacaat cctggatttt cttaagtccg atggatttgc
     3841 caaccggaac ttcatgcagt tgatccatga tgactctctc acctttaagg aggacatcca
     3901 gaaagcacaa gtttctggcc agggggacag tcttcacgag cacatcgcta atcttgcagg
     3961 tagcccagct atcaaaaagg gaatactgca gaccgttaag gtcgtggatg aactcgtcaa
     4021 agtaatggga aggcataagc ccgagaatat cgttatcgag atggcccgag agaaccaaac
     4081 tacccagaag ggacagaaga acagtaggga aaggatgaag aggattgaag agggtataaa
     4141 agaactgggg tcccaaatcc ttaaggaaca cccagttgaa aacacccagc ttcagaatga
     4201 gaagctctac ctgtactacc tgcagaacgg cagggacatg tacgtggatc aggaactgga
     4261 catcaatcgg ctctccgact acgacgtgga tcatatcgtg ccccagtctt ttctcaaaga
     4321 tgattctatt gataataaag tgttgacaag atccgataaa aatagaggga agagtgataa
     4381 cgtcccctca gaagaagttg tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc
     4441 caaactgatc acacaacgga agttcgataa tctgactaag gctgaacgag gtggcctgtc
     4501 tgagttggat aaagccggct tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa
     4561 gcacgtggcc caaattctcg attcacgcat gaacaccaag tacgatgaaa atgacaaact
     4621 gattcgagag gtgaaagtta ttactctgaa gtctaagctg gtctcagatt tcagaaagga
     4681 ctttcagttt tataaggtga gagagatcaa caattaccac catgcgcatg atgcctacct
     4741 gaatgcagtg gtaggcactg cacttatcaa aaaatatccc aagcttgaat ctgaatttgt
     4801 ttacggagac tataaagtgt acgatgttag gaaaatgatc gcaaagtctg agcaggaaat
     4861 aggcaaggcc accgctaagt acttctttta cagcaatatt atgaattttt tcaagaccga
     4921 gattacactg gccaatggag agattcggaa gcgaccactt atcgaaacaa acggagaaac
     4981 aggagaaatc gtgtgggaca agggtaggga tttcgcgaca gtccggaagg tcctgtccat
     5041 gccgcaggtg aacatcgtta aaaagaccga agtacagacc ggaggcttct ccaaggaaag
     5101 tatcctcccg aaaaggaaca gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa
     5161 gaaatacggc ggattcgatt ctcctacagt cgcttacagt gtactggttg tggccaaagt
     5221 ggagaaaggg aagtctaaaa aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat
     5281 ggagcgatca agcttcgaaa aaaaccccat cgactttctc gaggcgaaag gatataaaga
     5341 ggtcaaaaaa gacctcatca ttaagcttcc caagtactct ctctttgagc ttgaaaacgg
     5401 ccggaaacga atgctcgcta gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc
     5461 ctctaaatac gttaatttct tgtatctggc cagccactat gaaaagctca aagggtctcc
     5521 cgaagataat gagcagaagc agctgttcgt ggaacaacac aaacactacc ttgatgagat
     5581 catcgagcaa ataagcgaat tctccaaaag agtgatcctc gccgacgcta acctcgataa
     5641 ggtgctttct gcttacaata agcacaggga taagcccatc agggagcagg cagaaaacat
     5701 tatctacttg tttactctga ccaacttggg cgcgcctgca gccttcaagt acttcgacac
     5761 caccatagac agaaagcggt acacctctac aaaggaggtc ctggacgcca cactgattca
     5821 tcagtcaatt acggggctct atgaaacaag aatcgacctc tctcagctcg gtggagacag
     5881 cagggctgac cccaagaaga agaggaaggt gtgatctctt ctcgagtcat gtaattagtt
     5941 atgtcacgct tacgttcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt
     6001 agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg
     6061 ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt
     6121 atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcggccgg
     6181 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     6241 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     6301 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     6361 gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     6421 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     6481 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     6541 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     6601 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     6661 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     6721 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     6781 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg
     6841 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagggta ataactgata
     6901 taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt
     6961 tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg
     7021 ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat
     7081 gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc
     7141 cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc
     7201 acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc
     7261 aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa
     7321 catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac
     7381 aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta
     7441 tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc
     7501 gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat
     7561 ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc
     7621 ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt
     7681 ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc
     7741 acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg tttttgttct
     7801 gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat
     7861 atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga
     7921 atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt
     7981 gaaaaggtgg tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     8041 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     8101 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     8161 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     8221 atgataataa tggtttctta gtatgatcca atatcaaagg aaatgatagc attgaaggat
     8281 gagactaatc caattgagga gtggcagcat atagaacagc taaagggtag tgctgaagga
     8341 agcatacgat accccgcatg gaatgggata atatcacagg aggtactaga ctacctttca
     8401 tcctacataa atagacgcat ataagtacgc atttaagcat aaacacgcac tatgccgttc
     8461 ttctcatgta tatatatata caggcaacac gcagatatag gtgcgacgtg aacagtgagc
     8521 tgtatgtgcg cagctcgcgt tgcattttcg gaagcgctcg ttttcggaaa cgctttgaag
     8581 ttcctattcc gaagttccta ttctctagaa agtataggaa cttcagagcg cttttgaaaa
     8641 ccaaaagcgc tctgaagacg cactttcaaa aaaccaaaaa cgcaccggac tgtaacgagc
     8701 tactaaaata ttgcgaatac cgcttccaca aacattgctc aaaagtatct ctttgctata
     8761 tatctctgtg ctatatccct atataaccta cccatccacc tttcgctcct tgaacttgca
     8821 tctaaactcg acctctacat tttttatgtt tatctctagt attactcttt agacaaaaaa
     8881 attgtagtaa gaactattca tagagtgaat cgaaaacaat acgaaaatgt aaacatttcc
     8941 tatacgtagt atatagagac aaaatagaag aaaccgttca taattttctg accaatgaag
     9001 aatcatcaac gctatcactt tctgttcaca aagtatgcgc aatccacatc ggtatagaat
     9061 ataatcgggg atgcctttat cttgaaaaaa tgcacccgca gcttcgctag taatcagtaa
     9121 acgcgggaag tggagtcagg ctttttttat ggaagagaaa atagacacca aagtagcctt
     9181 cttctaacct taacggacct acagtgcaaa aagttatcaa gagactgcat tatagagcgc
     9241 acaaaggaga aaaaaagtaa tctaagatgc tttgttagaa aaatagcgct ctcgggatgc
     9301 atttttgtag aacaaaaaag aagtatagat tctttgttgg taaaatagcg ctctcgcgtt
     9361 gcatttctgt tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta gcgctctcgc
     9421 gttgcatttt tgttttacaa aaatgaagca cagattcttc gttggtaaaa tagcgctttc
     9481 gcgttgcatt tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa aattagcgct
     9541 ctcgcgttgc atttttgttc tacaaaatga agcacagatg cttcgttcag gtggcacttt
     9601 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     9661 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     9721 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     9781 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     9841 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     9901 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     9961 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
    10021 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
    10081 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
    10141 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
    10201 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
    10261 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
    10321 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
    10381 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
    10441 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
    10501 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
    10561 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
    10621 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
    10681 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
    10741 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
    10801 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
    10861 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
    10921 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
    10981 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
    11041 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
    11101 gcgaac
//
