LOCUS       sequence-193439-       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 ARS1014a sequence 
            TTATGTGCGTATTGCTTTCA in yeast chromosome 10..
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-ARS1014a
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     173..192
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     426..443
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    560..581
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(596..601,602..619,620..626)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    634..650
                     /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     639..661
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     658..674
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     658..674
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     693..713
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        695..713
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        940..1015
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1101..1805
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1815..5918
                     /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             5931..5951
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5967..6214
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6232..6251)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6233..6251)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6261..6278)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6261..6277)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6270..6292)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6418..6873
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6505..6524)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6715..6736
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7004..7807)
                     /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(7808..8023)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(8047..8066)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8166..8188
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8226..8244)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      8285..9627
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    8617..8664
                     /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        9654..9758
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9759..9827,9828..10619)
                     /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(9977..9996)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(10790..11106,1..272)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
       61 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
      121 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      181 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      241 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      301 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
      361 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
      421 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
      481 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
      541 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
      601 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
      661 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
      721 agctggagct agtatactct ttcttcaaca attaaatact ctcggtagcc aagttggttt
      781 aaggcgcaag actgtaattt atcactacga aatcttgaga tcgggcgttc gactcgcccc
      841 cgggagagat ggccggcatg gtcccagcct cctcgctggc gccggctggg caacaccttc
      901 gggtggcgaa tgggactttt tatgtgcgta ttgctttcag ttttagagct agaaatagca
      961 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     1021 ttattttttg tcactattgt tatgtaaaat gccacctctg acagtatgga acgcaaactt
     1081 ctgtctagtg gatagtcgac atccttttgt tgtttccggg tgtacaatat ggacttcctc
     1141 ttttctggca accaaaccca tacatcggga ttcctataat accttcgttg gtctccctaa
     1201 catgtaggtg gcggagggga gatatacaat agaacagata ccagacaaga cataatgggc
     1261 taaacaagac tacaccaatt acactgcctc attgatggtg gtacataacg aactaatact
     1321 gtagccctag acttgatagc catcatcata tcgaagtttc actacccttt ttccatttgc
     1381 catctattga agtaataata ggcgcatgca acttcttttc tttttttttc ttttctctct
     1441 cccccgttgt tgtctcacca tatccgcaat gacaaaaaaa tgatggaaga cactaaagga
     1501 aaaaattaac gacaaagaca gcaccaacag atgtcgttgt tccagagctg atgaggggta
     1561 tctcgaagca cacgaaactt tttccttcct tcattcacgc acactactct ctaatgagca
     1621 acggtatacg gccttccttc cagttacttg aatttgaaat aaaaaaaagt ttgctgtctt
     1681 gctatcaagt ataaatagac ctgcaattat taatcttttg tttcctcgtc attgttctcg
     1741 ttccctttct tccttgtttc tttttctgca caatatttca agctatacca agcatacaat
     1801 caacttggaa aaaaatggac aagaagtact ccattgggct cgatatcggc acaaacagcg
     1861 tcggttgggc cgtcattacg gacgagtaca aggtgccgag caaaaaattc aaagttctgg
     1921 gcaataccga tcgccacagc ataaagaaga acctcattgg cgccctcctg ttcgactccg
     1981 gggagacggc cgaagccacg cggctcaaaa gaacagcacg gcgcagatat acccgcagaa
     2041 agaatcggat ctgctacctg caggagatct ttagtaatga gatggctaag gtggatgact
     2101 ctttcttcca taggctggag gagtcctttt tggtggagga ggataaaaag aacgagcgcc
     2161 acccaatctt tggcaatatc gtggacgagg tggcgtacca tgaaaagtac ccaaccatat
     2221 atcatctgag gaagaagctt gtagacagta ctgataaggc tgacttgcgg ttgatctatc
     2281 tcgcgctggc gcatatgatc aaatttcggg gacacttcct catcgagggg gacctgaacc
     2341 cagacaacag cgatgtcgac aaactcttta tccaactggt tcagacttac aatcagcttt
     2401 tcgaagagaa cccgatcaac gcatccggag ttgacgccaa agcaatcctg agcgctaggc
     2461 tgtccaaatc ccggcggctc gaaaacctca tcgcacagct ccctggggag aagaagaacg
     2521 gcctgtttgg taatcttatc gccctgtcac tcgggctgac ccccaacttt aaatctaact
     2581 tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga cacctacgat gatgatctcg
     2641 acaatctgct ggcccagatc ggcgaccagt acgcagacct ttttttggcg gcaaagaacc
     2701 tgtcagacgc cattctgctg agtgatattc tgcgagtgaa cacggagatc accaaagctc
     2761 cgctgagcgc tagtatgatc aagcgctatg atgagcacca ccaagacttg actttgctga
     2821 aggcccttgt cagacagcaa ctgcctgaga agtacaagga aattttcttc gatcagtcta
     2881 aaaatggcta cgccggatac attgacggcg gagcaagcca ggaggaattt tacaaattta
     2941 ttaagcccat cttggaaaaa atggacggca ccgaggagct gctggtaaag cttaacagag
     3001 aagatctgtt gcgcaaacag cgcactttcg acaatggaag catcccccac cagattcacc
     3061 tgggcgaact gcacgctatc ctcaggcggc aagaggattt ctaccccttt ttgaaagata
     3121 acagggaaaa gattgagaaa atcctcacat ttcggatacc ctactatgta ggccccctcg
     3181 cccggggaaa ttccagattc gcgtggatga ctcgcaaatc agaagagacc atcactccct
     3241 ggaacttcga ggaagtcgtg gataaggggg cctctgccca gtccttcatc gaaaggatga
     3301 ctaactttga taaaaatctg cctaacgaaa aggtgcttcc taaacactct ctgctgtacg
     3361 agtacttcac agtttataac gagctcacca aggtcaaata cgtcacagaa gggatgagaa
     3421 agccagcatt cctgtctgga gagcagaaga aagctatcgt ggacctcctc ttcaagacga
     3481 accggaaagt taccgtgaaa cagctcaaag aagactattt caaaaagatt gaatgtttcg
     3541 actctgttga aatcagcgga gtggaggatc gcttcaacgc atccctggga acgtatcacg
     3601 atctcctgaa aatcattaaa gacaaggact tcctggacaa tgaggagaac gaggacattc
     3661 ttgaggacat tgtcctcacc cttacgttgt ttgaagatag ggagatgatt gaagaacgct
     3721 tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa acagctcaag aggcgccgat
     3781 atacaggatg ggggcggctg tcaagaaaac tgatcaatgg gatccgagac aagcagagtg
     3841 gaaagacaat cctggatttt cttaagtccg atggatttgc caaccggaac ttcatgcagt
     3901 tgatccatga tgactctctc acctttaagg aggacatcca gaaagcacaa gtttctggcc
     3961 agggggacag tcttcacgag cacatcgcta atcttgcagg tagcccagct atcaaaaagg
     4021 gaatactgca gaccgttaag gtcgtggatg aactcgtcaa agtaatggga aggcataagc
     4081 ccgagaatat cgttatcgag atggcccgag agaaccaaac tacccagaag ggacagaaga
     4141 acagtaggga aaggatgaag aggattgaag agggtataaa agaactgggg tcccaaatcc
     4201 ttaaggaaca cccagttgaa aacacccagc ttcagaatga gaagctctac ctgtactacc
     4261 tgcagaacgg cagggacatg tacgtggatc aggaactgga catcaatcgg ctctccgact
     4321 acgacgtgga tcatatcgtg ccccagtctt ttctcaaaga tgattctatt gataataaag
     4381 tgttgacaag atccgataaa aatagaggga agagtgataa cgtcccctca gaagaagttg
     4441 tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc caaactgatc acacaacgga
     4501 agttcgataa tctgactaag gctgaacgag gtggcctgtc tgagttggat aaagccggct
     4561 tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa gcacgtggcc caaattctcg
     4621 attcacgcat gaacaccaag tacgatgaaa atgacaaact gattcgagag gtgaaagtta
     4681 ttactctgaa gtctaagctg gtctcagatt tcagaaagga ctttcagttt tataaggtga
     4741 gagagatcaa caattaccac catgcgcatg atgcctacct gaatgcagtg gtaggcactg
     4801 cacttatcaa aaaatatccc aagcttgaat ctgaatttgt ttacggagac tataaagtgt
     4861 acgatgttag gaaaatgatc gcaaagtctg agcaggaaat aggcaaggcc accgctaagt
     4921 acttctttta cagcaatatt atgaattttt tcaagaccga gattacactg gccaatggag
     4981 agattcggaa gcgaccactt atcgaaacaa acggagaaac aggagaaatc gtgtgggaca
     5041 agggtaggga tttcgcgaca gtccggaagg tcctgtccat gccgcaggtg aacatcgtta
     5101 aaaagaccga agtacagacc ggaggcttct ccaaggaaag tatcctcccg aaaaggaaca
     5161 gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa gaaatacggc ggattcgatt
     5221 ctcctacagt cgcttacagt gtactggttg tggccaaagt ggagaaaggg aagtctaaaa
     5281 aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat ggagcgatca agcttcgaaa
     5341 aaaaccccat cgactttctc gaggcgaaag gatataaaga ggtcaaaaaa gacctcatca
     5401 ttaagcttcc caagtactct ctctttgagc ttgaaaacgg ccggaaacga atgctcgcta
     5461 gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc ctctaaatac gttaatttct
     5521 tgtatctggc cagccactat gaaaagctca aagggtctcc cgaagataat gagcagaagc
     5581 agctgttcgt ggaacaacac aaacactacc ttgatgagat catcgagcaa ataagcgaat
     5641 tctccaaaag agtgatcctc gccgacgcta acctcgataa ggtgctttct gcttacaata
     5701 agcacaggga taagcccatc agggagcagg cagaaaacat tatctacttg tttactctga
     5761 ccaacttggg cgcgcctgca gccttcaagt acttcgacac caccatagac agaaagcggt
     5821 acacctctac aaaggaggtc ctggacgcca cactgattca tcagtcaatt acggggctct
     5881 atgaaacaag aatcgacctc tctcagctcg gtggagacag cagggctgac cccaagaaga
     5941 agaggaaggt gtgatctctt ctcgagtcat gtaattagtt atgtcacgct tacgttcacg
     6001 ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
     6061 ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt
     6121 tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga
     6181 gaaggttttg ggacgctcga aggctttaat ttgcggccgg tacccaattc gccctatagt
     6241 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     6301 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     6361 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg
     6421 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     6481 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     6541 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     6601 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     6661 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     6721 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     6781 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     6841 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     6901 tctgtgcggt atttcacacc gcatagggta ataactgata taattaaatt gaagctctaa
     6961 tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt tgctggccgc
     7021 atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct accttagcat
     7081 cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct gtagagacca
     7141 catcatccac ggttctatac tgttgaccca atgcgtctcc cttgtcatct aaacccacac
     7201 cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct ctttgagcaa
     7261 taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt
     7321 ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg cctctaggtt
     7381 cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg cccaccacac
     7441 cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca gagtactgca
     7501 atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact
     7561 tggcggataa tgcctttagc ggcttaactg tgccctccat ggaaaaatca gtcaagatat
     7621 ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac tccagtaatt
     7681 ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg tgcatgatat
     7741 taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta tatgtagctt
     7801 tcgacatgat ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg gttaagaata
     7861 ctgggcaatt tcatgtttct tcaacactac atatgcgtat atataccaat ctaagtctgt
     7921 gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa
     7981 accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaaaaggtgg tatggtgcac
     8041 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     8101 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     8161 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     8221 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     8281 gtatgatcca atatcaaagg aaatgatagc attgaaggat gagactaatc caattgagga
     8341 gtggcagcat atagaacagc taaagggtag tgctgaagga agcatacgat accccgcatg
     8401 gaatgggata atatcacagg aggtactaga ctacctttca tcctacataa atagacgcat
     8461 ataagtacgc atttaagcat aaacacgcac tatgccgttc ttctcatgta tatatatata
     8521 caggcaacac gcagatatag gtgcgacgtg aacagtgagc tgtatgtgcg cagctcgcgt
     8581 tgcattttcg gaagcgctcg ttttcggaaa cgctttgaag ttcctattcc gaagttccta
     8641 ttctctagaa agtataggaa cttcagagcg cttttgaaaa ccaaaagcgc tctgaagacg
     8701 cactttcaaa aaaccaaaaa cgcaccggac tgtaacgagc tactaaaata ttgcgaatac
     8761 cgcttccaca aacattgctc aaaagtatct ctttgctata tatctctgtg ctatatccct
     8821 atataaccta cccatccacc tttcgctcct tgaacttgca tctaaactcg acctctacat
     8881 tttttatgtt tatctctagt attactcttt agacaaaaaa attgtagtaa gaactattca
     8941 tagagtgaat cgaaaacaat acgaaaatgt aaacatttcc tatacgtagt atatagagac
     9001 aaaatagaag aaaccgttca taattttctg accaatgaag aatcatcaac gctatcactt
     9061 tctgttcaca aagtatgcgc aatccacatc ggtatagaat ataatcgggg atgcctttat
     9121 cttgaaaaaa tgcacccgca gcttcgctag taatcagtaa acgcgggaag tggagtcagg
     9181 ctttttttat ggaagagaaa atagacacca aagtagcctt cttctaacct taacggacct
     9241 acagtgcaaa aagttatcaa gagactgcat tatagagcgc acaaaggaga aaaaaagtaa
     9301 tctaagatgc tttgttagaa aaatagcgct ctcgggatgc atttttgtag aacaaaaaag
     9361 aagtatagat tctttgttgg taaaatagcg ctctcgcgtt gcatttctgt tctgtaaaaa
     9421 tgcagctcag attctttgtt tgaaaaatta gcgctctcgc gttgcatttt tgttttacaa
     9481 aaatgaagca cagattcttc gttggtaaaa tagcgctttc gcgttgcatt tctgttctgt
     9541 aaaaatgcag ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc atttttgttc
     9601 tacaaaatga agcacagatg cttcgttcag gtggcacttt tcggggaaat gtgcgcggaa
     9661 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     9721 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     9781 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     9841 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     9901 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     9961 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
    10021 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
    10081 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
    10141 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
    10201 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
    10261 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
    10321 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
    10381 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
    10441 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
    10501 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
    10561 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
    10621 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
    10681 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
    10741 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
    10801 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
    10861 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
    10921 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
    10981 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
    11041 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
    11101 cgggct
//
