LOCUS       sequence-193424-       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 ARS106a sequence 
            ATACGGTCAGGGTAGCGCCC in yeast chromosome 1..
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-ARS106a
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     81..100
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     334..351
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    468..489
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(504..509,510..527,528..534)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    542..558
                     /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     547..569
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     566..582
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     566..582
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     601..621
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        603..621
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        848..923
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1009..1713
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1723..5826
                     /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             5839..5859
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5875..6122
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6140..6159)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6141..6159)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6169..6186)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6169..6185)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6178..6200)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6326..6781
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6413..6432)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6623..6644
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6912..7715)
                     /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="MSKATYKERAATHPSPVAAKLFNIMHEKQTNSCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(7716..7931)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7955..7974)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8074..8096
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8134..8152)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      8193..9535
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    8525..8572
                     /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        9562..9666
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9667..9735,9736..10527)
                     /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(9885..9904)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(10698..11106,1..180)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
       61 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      121 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      181 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      241 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      301 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      361 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      421 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      481 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      541 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
      601 gcaattaacc ctcactaaag ggaacaaaag ctggagctag tatactcttt cttcaacaat
      661 taaatactct cggtagccaa gttggtttaa ggcgcaagac tgtaatttat cactacgaaa
      721 tcttgagatc gggcgttcga ctcgcccccg ggagagatgg ccggcatggt cccagcctcc
      781 tcgctggcgc cggctgggca acaccttcgg gtggcgaatg ggactttata cggtcagggt
      841 agcgcccgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
      901 aaaaagtggc accgagtcgg tgcttttttt attttttgtc actattgtta tgtaaaatgc
      961 cacctctgac agtatggaac gcaaacttct gtctagtgga tagtcgacat ccttttgttg
     1021 tttccgggtg tacaatatgg acttcctctt ttctggcaac caaacccata catcgggatt
     1081 cctataatac cttcgttggt ctccctaaca tgtaggtggc ggaggggaga tatacaatag
     1141 aacagatacc agacaagaca taatgggcta aacaagacta caccaattac actgcctcat
     1201 tgatggtggt acataacgaa ctaatactgt agccctagac ttgatagcca tcatcatatc
     1261 gaagtttcac tacccttttt ccatttgcca tctattgaag taataatagg cgcatgcaac
     1321 ttcttttctt tttttttctt ttctctctcc cccgttgttg tctcaccata tccgcaatga
     1381 caaaaaaatg atggaagaca ctaaaggaaa aaattaacga caaagacagc accaacagat
     1441 gtcgttgttc cagagctgat gaggggtatc tcgaagcaca cgaaactttt tccttccttc
     1501 attcacgcac actactctct aatgagcaac ggtatacggc cttccttcca gttacttgaa
     1561 tttgaaataa aaaaaagttt gctgtcttgc tatcaagtat aaatagacct gcaattatta
     1621 atcttttgtt tcctcgtcat tgttctcgtt ccctttcttc cttgtttctt tttctgcaca
     1681 atatttcaag ctataccaag catacaatca acttggaaaa aaatggacaa gaagtactcc
     1741 attgggctcg atatcggcac aaacagcgtc ggttgggccg tcattacgga cgagtacaag
     1801 gtgccgagca aaaaattcaa agttctgggc aataccgatc gccacagcat aaagaagaac
     1861 ctcattggcg ccctcctgtt cgactccggg gagacggccg aagccacgcg gctcaaaaga
     1921 acagcacggc gcagatatac ccgcagaaag aatcggatct gctacctgca ggagatcttt
     1981 agtaatgaga tggctaaggt ggatgactct ttcttccata ggctggagga gtcctttttg
     2041 gtggaggagg ataaaaagaa cgagcgccac ccaatctttg gcaatatcgt ggacgaggtg
     2101 gcgtaccatg aaaagtaccc aaccatatat catctgagga agaagcttgt agacagtact
     2161 gataaggctg acttgcggtt gatctatctc gcgctggcgc atatgatcaa atttcgggga
     2221 cacttcctca tcgaggggga cctgaaccca gacaacagcg atgtcgacaa actctttatc
     2281 caactggttc agacttacaa tcagcttttc gaagagaacc cgatcaacgc atccggagtt
     2341 gacgccaaag caatcctgag cgctaggctg tccaaatccc ggcggctcga aaacctcatc
     2401 gcacagctcc ctggggagaa gaagaacggc ctgtttggta atcttatcgc cctgtcactc
     2461 gggctgaccc ccaactttaa atctaacttc gacctggccg aagatgccaa gcttcaactg
     2521 agcaaagaca cctacgatga tgatctcgac aatctgctgg cccagatcgg cgaccagtac
     2581 gcagaccttt ttttggcggc aaagaacctg tcagacgcca ttctgctgag tgatattctg
     2641 cgagtgaaca cggagatcac caaagctccg ctgagcgcta gtatgatcaa gcgctatgat
     2701 gagcaccacc aagacttgac tttgctgaag gcccttgtca gacagcaact gcctgagaag
     2761 tacaaggaaa ttttcttcga tcagtctaaa aatggctacg ccggatacat tgacggcgga
     2821 gcaagccagg aggaatttta caaatttatt aagcccatct tggaaaaaat ggacggcacc
     2881 gaggagctgc tggtaaagct taacagagaa gatctgttgc gcaaacagcg cactttcgac
     2941 aatggaagca tcccccacca gattcacctg ggcgaactgc acgctatcct caggcggcaa
     3001 gaggatttct accccttttt gaaagataac agggaaaaga ttgagaaaat cctcacattt
     3061 cggataccct actatgtagg ccccctcgcc cggggaaatt ccagattcgc gtggatgact
     3121 cgcaaatcag aagagaccat cactccctgg aacttcgagg aagtcgtgga taagggggcc
     3181 tctgcccagt ccttcatcga aaggatgact aactttgata aaaatctgcc taacgaaaag
     3241 gtgcttccta aacactctct gctgtacgag tacttcacag tttataacga gctcaccaag
     3301 gtcaaatacg tcacagaagg gatgagaaag ccagcattcc tgtctggaga gcagaagaaa
     3361 gctatcgtgg acctcctctt caagacgaac cggaaagtta ccgtgaaaca gctcaaagaa
     3421 gactatttca aaaagattga atgtttcgac tctgttgaaa tcagcggagt ggaggatcgc
     3481 ttcaacgcat ccctgggaac gtatcacgat ctcctgaaaa tcattaaaga caaggacttc
     3541 ctggacaatg aggagaacga ggacattctt gaggacattg tcctcaccct tacgttgttt
     3601 gaagataggg agatgattga agaacgcttg aaaacttacg ctcatctctt cgacgacaaa
     3661 gtcatgaaac agctcaagag gcgccgatat acaggatggg ggcggctgtc aagaaaactg
     3721 atcaatggga tccgagacaa gcagagtgga aagacaatcc tggattttct taagtccgat
     3781 ggatttgcca accggaactt catgcagttg atccatgatg actctctcac ctttaaggag
     3841 gacatccaga aagcacaagt ttctggccag ggggacagtc ttcacgagca catcgctaat
     3901 cttgcaggta gcccagctat caaaaaggga atactgcaga ccgttaaggt cgtggatgaa
     3961 ctcgtcaaag taatgggaag gcataagccc gagaatatcg ttatcgagat ggcccgagag
     4021 aaccaaacta cccagaaggg acagaagaac agtagggaaa ggatgaagag gattgaagag
     4081 ggtataaaag aactggggtc ccaaatcctt aaggaacacc cagttgaaaa cacccagctt
     4141 cagaatgaga agctctacct gtactacctg cagaacggca gggacatgta cgtggatcag
     4201 gaactggaca tcaatcggct ctccgactac gacgtggatc atatcgtgcc ccagtctttt
     4261 ctcaaagatg attctattga taataaagtg ttgacaagat ccgataaaaa tagagggaag
     4321 agtgataacg tcccctcaga agaagttgtc aagaaaatga aaaattattg gcggcagctg
     4381 ctgaacgcca aactgatcac acaacggaag ttcgataatc tgactaaggc tgaacgaggt
     4441 ggcctgtctg agttggataa agccggcttc atcaaaaggc agcttgttga gacacgccag
     4501 atcaccaagc acgtggccca aattctcgat tcacgcatga acaccaagta cgatgaaaat
     4561 gacaaactga ttcgagaggt gaaagttatt actctgaagt ctaagctggt ctcagatttc
     4621 agaaaggact ttcagtttta taaggtgaga gagatcaaca attaccacca tgcgcatgat
     4681 gcctacctga atgcagtggt aggcactgca cttatcaaaa aatatcccaa gcttgaatct
     4741 gaatttgttt acggagacta taaagtgtac gatgttagga aaatgatcgc aaagtctgag
     4801 caggaaatag gcaaggccac cgctaagtac ttcttttaca gcaatattat gaattttttc
     4861 aagaccgaga ttacactggc caatggagag attcggaagc gaccacttat cgaaacaaac
     4921 ggagaaacag gagaaatcgt gtgggacaag ggtagggatt tcgcgacagt ccggaaggtc
     4981 ctgtccatgc cgcaggtgaa catcgttaaa aagaccgaag tacagaccgg aggcttctcc
     5041 aaggaaagta tcctcccgaa aaggaacagc gacaagctga tcgcacgcaa aaaagattgg
     5101 gaccccaaga aatacggcgg attcgattct cctacagtcg cttacagtgt actggttgtg
     5161 gccaaagtgg agaaagggaa gtctaaaaaa ctcaaaagcg tcaaggaact gctgggcatc
     5221 acaatcatgg agcgatcaag cttcgaaaaa aaccccatcg actttctcga ggcgaaagga
     5281 tataaagagg tcaaaaaaga cctcatcatt aagcttccca agtactctct ctttgagctt
     5341 gaaaacggcc ggaaacgaat gctcgctagt gcgggcgagc tgcagaaagg taacgagctg
     5401 gcactgccct ctaaatacgt taatttcttg tatctggcca gccactatga aaagctcaaa
     5461 gggtctcccg aagataatga gcagaagcag ctgttcgtgg aacaacacaa acactacctt
     5521 gatgagatca tcgagcaaat aagcgaattc tccaaaagag tgatcctcgc cgacgctaac
     5581 ctcgataagg tgctttctgc ttacaataag cacagggata agcccatcag ggagcaggca
     5641 gaaaacatta tctacttgtt tactctgacc aacttgggcg cgcctgcagc cttcaagtac
     5701 ttcgacacca ccatagacag aaagcggtac acctctacaa aggaggtcct ggacgccaca
     5761 ctgattcatc agtcaattac ggggctctat gaaacaagaa tcgacctctc tcagctcggt
     5821 ggagacagca gggctgaccc caagaagaag aggaaggtgt gatctcttct cgagtcatgt
     5881 aattagttat gtcacgctta cgttcacgcc ctccccccac atccgctcta accgaaaagg
     5941 aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat
     6001 taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat
     6061 gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt
     6121 gcggccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     6181 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     6241 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     6301 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     6361 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     6421 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     6481 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     6541 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     6601 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     6661 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     6721 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     6781 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtaat
     6841 aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat
     6901 aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt
     6961 ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca
     7021 ataataatgt cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat
     7081 gcgtctccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca
     7141 tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc
     7201 gcaatgtcaa cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat
     7261 tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa
     7321 ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct
     7381 attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt
     7441 ctgtcttcga agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg
     7501 ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga
     7561 cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac
     7621 gagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga
     7681 gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt cctgcaggtt
     7741 tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat
     7801 atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag
     7861 attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat
     7921 gatgaattga aaaggtggta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     7981 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     8041 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     8101 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     8161 ttaatgtcat gataataatg gtttcttagt atgatccaat atcaaaggaa atgatagcat
     8221 tgaaggatga gactaatcca attgaggagt ggcagcatat agaacagcta aagggtagtg
     8281 ctgaaggaag catacgatac cccgcatgga atgggataat atcacaggag gtactagact
     8341 acctttcatc ctacataaat agacgcatat aagtacgcat ttaagcataa acacgcacta
     8401 tgccgttctt ctcatgtata tatatataca ggcaacacgc agatataggt gcgacgtgaa
     8461 cagtgagctg tatgtgcgca gctcgcgttg cattttcgga agcgctcgtt ttcggaaacg
     8521 ctttgaagtt cctattccga agttcctatt ctctagaaag tataggaact tcagagcgct
     8581 tttgaaaacc aaaagcgctc tgaagacgca ctttcaaaaa accaaaaacg caccggactg
     8641 taacgagcta ctaaaatatt gcgaataccg cttccacaaa cattgctcaa aagtatctct
     8701 ttgctatata tctctgtgct atatccctat ataacctacc catccacctt tcgctccttg
     8761 aacttgcatc taaactcgac ctctacattt tttatgttta tctctagtat tactctttag
     8821 acaaaaaaat tgtagtaaga actattcata gagtgaatcg aaaacaatac gaaaatgtaa
     8881 acatttccta tacgtagtat atagagacaa aatagaagaa accgttcata attttctgac
     8941 caatgaagaa tcatcaacgc tatcactttc tgttcacaaa gtatgcgcaa tccacatcgg
     9001 tatagaatat aatcggggat gcctttatct tgaaaaaatg cacccgcagc ttcgctagta
     9061 atcagtaaac gcgggaagtg gagtcaggct ttttttatgg aagagaaaat agacaccaaa
     9121 gtagccttct tctaacctta acggacctac agtgcaaaaa gttatcaaga gactgcatta
     9181 tagagcgcac aaaggagaaa aaaagtaatc taagatgctt tgttagaaaa atagcgctct
     9241 cgggatgcat ttttgtagaa caaaaaagaa gtatagattc tttgttggta aaatagcgct
     9301 ctcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc
     9361 gctctcgcgt tgcatttttg ttttacaaaa atgaagcaca gattcttcgt tggtaaaata
     9421 gcgctttcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa
     9481 ttagcgctct cgcgttgcat ttttgttcta caaaatgaag cacagatgct tcgttcaggt
     9541 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     9601 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     9661 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     9721 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     9781 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     9841 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     9901 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     9961 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
    10021 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
    10081 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
    10141 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
    10201 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
    10261 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
    10321 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
    10381 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
    10441 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
    10501 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
    10561 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
    10621 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
    10681 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
    10741 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
    10801 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
    10861 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
    10921 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
    10981 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
    11041 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
    11101 gccacg
//
