LOCUS       sequence-193436-       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 ARS805a sequence 
            TTATTTGAATGATATTTAGT in yeast chromosome 8..
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-ARS805a
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     71..90
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     324..341
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    458..479
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(494..499,500..517,518..524)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    532..548
                     /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     537..559
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     556..572
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     556..572
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     591..611
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        593..611
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        838..913
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        999..1703
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1713..5816
                     /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             5829..5849
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5865..6112
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6130..6149)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6131..6149)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6159..6176)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6159..6175)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6168..6190)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6316..6771
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6403..6422)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6613..6634
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6902..7705)
                     /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(7706..7921)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7945..7964)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8064..8086
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8124..8142)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      8183..9525
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    8515..8562
                     /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        9552..9656
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9657..9725,9726..10517)
                     /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(9875..9894)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(10688..11106,1..170)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
       61 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      121 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      181 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      241 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
      301 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
      361 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
      421 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
      481 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
      541 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
      601 ctcactaaag ggaacaaaag ctggagctag tatactcttt cttcaacaat taaatactct
      661 cggtagccaa gttggtttaa ggcgcaagac tgtaatttat cactacgaaa tcttgagatc
      721 gggcgttcga ctcgcccccg ggagagatgg ccggcatggt cccagcctcc tcgctggcgc
      781 cggctgggca acaccttcgg gtggcgaatg ggacttttta tttgaatgat atttagtgtt
      841 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
      901 accgagtcgg tgcttttttt attttttgtc actattgtta tgtaaaatgc cacctctgac
      961 agtatggaac gcaaacttct gtctagtgga tagtcgacat ccttttgttg tttccgggtg
     1021 tacaatatgg acttcctctt ttctggcaac caaacccata catcgggatt cctataatac
     1081 cttcgttggt ctccctaaca tgtaggtggc ggaggggaga tatacaatag aacagatacc
     1141 agacaagaca taatgggcta aacaagacta caccaattac actgcctcat tgatggtggt
     1201 acataacgaa ctaatactgt agccctagac ttgatagcca tcatcatatc gaagtttcac
     1261 tacccttttt ccatttgcca tctattgaag taataatagg cgcatgcaac ttcttttctt
     1321 tttttttctt ttctctctcc cccgttgttg tctcaccata tccgcaatga caaaaaaatg
     1381 atggaagaca ctaaaggaaa aaattaacga caaagacagc accaacagat gtcgttgttc
     1441 cagagctgat gaggggtatc tcgaagcaca cgaaactttt tccttccttc attcacgcac
     1501 actactctct aatgagcaac ggtatacggc cttccttcca gttacttgaa tttgaaataa
     1561 aaaaaagttt gctgtcttgc tatcaagtat aaatagacct gcaattatta atcttttgtt
     1621 tcctcgtcat tgttctcgtt ccctttcttc cttgtttctt tttctgcaca atatttcaag
     1681 ctataccaag catacaatca acttggaaaa aaatggacaa gaagtactcc attgggctcg
     1741 atatcggcac aaacagcgtc ggttgggccg tcattacgga cgagtacaag gtgccgagca
     1801 aaaaattcaa agttctgggc aataccgatc gccacagcat aaagaagaac ctcattggcg
     1861 ccctcctgtt cgactccggg gagacggccg aagccacgcg gctcaaaaga acagcacggc
     1921 gcagatatac ccgcagaaag aatcggatct gctacctgca ggagatcttt agtaatgaga
     1981 tggctaaggt ggatgactct ttcttccata ggctggagga gtcctttttg gtggaggagg
     2041 ataaaaagaa cgagcgccac ccaatctttg gcaatatcgt ggacgaggtg gcgtaccatg
     2101 aaaagtaccc aaccatatat catctgagga agaagcttgt agacagtact gataaggctg
     2161 acttgcggtt gatctatctc gcgctggcgc atatgatcaa atttcgggga cacttcctca
     2221 tcgaggggga cctgaaccca gacaacagcg atgtcgacaa actctttatc caactggttc
     2281 agacttacaa tcagcttttc gaagagaacc cgatcaacgc atccggagtt gacgccaaag
     2341 caatcctgag cgctaggctg tccaaatccc ggcggctcga aaacctcatc gcacagctcc
     2401 ctggggagaa gaagaacggc ctgtttggta atcttatcgc cctgtcactc gggctgaccc
     2461 ccaactttaa atctaacttc gacctggccg aagatgccaa gcttcaactg agcaaagaca
     2521 cctacgatga tgatctcgac aatctgctgg cccagatcgg cgaccagtac gcagaccttt
     2581 ttttggcggc aaagaacctg tcagacgcca ttctgctgag tgatattctg cgagtgaaca
     2641 cggagatcac caaagctccg ctgagcgcta gtatgatcaa gcgctatgat gagcaccacc
     2701 aagacttgac tttgctgaag gcccttgtca gacagcaact gcctgagaag tacaaggaaa
     2761 ttttcttcga tcagtctaaa aatggctacg ccggatacat tgacggcgga gcaagccagg
     2821 aggaatttta caaatttatt aagcccatct tggaaaaaat ggacggcacc gaggagctgc
     2881 tggtaaagct taacagagaa gatctgttgc gcaaacagcg cactttcgac aatggaagca
     2941 tcccccacca gattcacctg ggcgaactgc acgctatcct caggcggcaa gaggatttct
     3001 accccttttt gaaagataac agggaaaaga ttgagaaaat cctcacattt cggataccct
     3061 actatgtagg ccccctcgcc cggggaaatt ccagattcgc gtggatgact cgcaaatcag
     3121 aagagaccat cactccctgg aacttcgagg aagtcgtgga taagggggcc tctgcccagt
     3181 ccttcatcga aaggatgact aactttgata aaaatctgcc taacgaaaag gtgcttccta
     3241 aacactctct gctgtacgag tacttcacag tttataacga gctcaccaag gtcaaatacg
     3301 tcacagaagg gatgagaaag ccagcattcc tgtctggaga gcagaagaaa gctatcgtgg
     3361 acctcctctt caagacgaac cggaaagtta ccgtgaaaca gctcaaagaa gactatttca
     3421 aaaagattga atgtttcgac tctgttgaaa tcagcggagt ggaggatcgc ttcaacgcat
     3481 ccctgggaac gtatcacgat ctcctgaaaa tcattaaaga caaggacttc ctggacaatg
     3541 aggagaacga ggacattctt gaggacattg tcctcaccct tacgttgttt gaagataggg
     3601 agatgattga agaacgcttg aaaacttacg ctcatctctt cgacgacaaa gtcatgaaac
     3661 agctcaagag gcgccgatat acaggatggg ggcggctgtc aagaaaactg atcaatggga
     3721 tccgagacaa gcagagtgga aagacaatcc tggattttct taagtccgat ggatttgcca
     3781 accggaactt catgcagttg atccatgatg actctctcac ctttaaggag gacatccaga
     3841 aagcacaagt ttctggccag ggggacagtc ttcacgagca catcgctaat cttgcaggta
     3901 gcccagctat caaaaaggga atactgcaga ccgttaaggt cgtggatgaa ctcgtcaaag
     3961 taatgggaag gcataagccc gagaatatcg ttatcgagat ggcccgagag aaccaaacta
     4021 cccagaaggg acagaagaac agtagggaaa ggatgaagag gattgaagag ggtataaaag
     4081 aactggggtc ccaaatcctt aaggaacacc cagttgaaaa cacccagctt cagaatgaga
     4141 agctctacct gtactacctg cagaacggca gggacatgta cgtggatcag gaactggaca
     4201 tcaatcggct ctccgactac gacgtggatc atatcgtgcc ccagtctttt ctcaaagatg
     4261 attctattga taataaagtg ttgacaagat ccgataaaaa tagagggaag agtgataacg
     4321 tcccctcaga agaagttgtc aagaaaatga aaaattattg gcggcagctg ctgaacgcca
     4381 aactgatcac acaacggaag ttcgataatc tgactaaggc tgaacgaggt ggcctgtctg
     4441 agttggataa agccggcttc atcaaaaggc agcttgttga gacacgccag atcaccaagc
     4501 acgtggccca aattctcgat tcacgcatga acaccaagta cgatgaaaat gacaaactga
     4561 ttcgagaggt gaaagttatt actctgaagt ctaagctggt ctcagatttc agaaaggact
     4621 ttcagtttta taaggtgaga gagatcaaca attaccacca tgcgcatgat gcctacctga
     4681 atgcagtggt aggcactgca cttatcaaaa aatatcccaa gcttgaatct gaatttgttt
     4741 acggagacta taaagtgtac gatgttagga aaatgatcgc aaagtctgag caggaaatag
     4801 gcaaggccac cgctaagtac ttcttttaca gcaatattat gaattttttc aagaccgaga
     4861 ttacactggc caatggagag attcggaagc gaccacttat cgaaacaaac ggagaaacag
     4921 gagaaatcgt gtgggacaag ggtagggatt tcgcgacagt ccggaaggtc ctgtccatgc
     4981 cgcaggtgaa catcgttaaa aagaccgaag tacagaccgg aggcttctcc aaggaaagta
     5041 tcctcccgaa aaggaacagc gacaagctga tcgcacgcaa aaaagattgg gaccccaaga
     5101 aatacggcgg attcgattct cctacagtcg cttacagtgt actggttgtg gccaaagtgg
     5161 agaaagggaa gtctaaaaaa ctcaaaagcg tcaaggaact gctgggcatc acaatcatgg
     5221 agcgatcaag cttcgaaaaa aaccccatcg actttctcga ggcgaaagga tataaagagg
     5281 tcaaaaaaga cctcatcatt aagcttccca agtactctct ctttgagctt gaaaacggcc
     5341 ggaaacgaat gctcgctagt gcgggcgagc tgcagaaagg taacgagctg gcactgccct
     5401 ctaaatacgt taatttcttg tatctggcca gccactatga aaagctcaaa gggtctcccg
     5461 aagataatga gcagaagcag ctgttcgtgg aacaacacaa acactacctt gatgagatca
     5521 tcgagcaaat aagcgaattc tccaaaagag tgatcctcgc cgacgctaac ctcgataagg
     5581 tgctttctgc ttacaataag cacagggata agcccatcag ggagcaggca gaaaacatta
     5641 tctacttgtt tactctgacc aacttgggcg cgcctgcagc cttcaagtac ttcgacacca
     5701 ccatagacag aaagcggtac acctctacaa aggaggtcct ggacgccaca ctgattcatc
     5761 agtcaattac ggggctctat gaaacaagaa tcgacctctc tcagctcggt ggagacagca
     5821 gggctgaccc caagaagaag aggaaggtgt gatctcttct cgagtcatgt aattagttat
     5881 gtcacgctta cgttcacgcc ctccccccac atccgctcta accgaaaagg aaggagttag
     5941 acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt
     6001 atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat
     6061 actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt gcggccggta
     6121 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     6181 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     6241 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     6301 ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     6361 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     6421 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     6481 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     6541 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     6601 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     6661 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     6721 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     6781 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtaat aactgatata
     6841 attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat aatacagttt
     6901 tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt ctgtaacgtt
     6961 caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca ataataatgt
     7021 cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat gcgtctccct
     7081 tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca tctcttccac
     7141 ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa
     7201 cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat tctgctaaca
     7261 tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa
     7321 tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct attctgtata
     7381 cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt ctgtcttcga
     7441 agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg ccctccatgg
     7501 aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga cctaatgctt
     7561 caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac aagtttgttt
     7621 gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga gtagcagcac
     7681 gttccttata tgtagctttc gacatgattt atcttcgttt cctgcaggtt tttgttctgt
     7741 gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat atgcgtatat
     7801 ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat
     7861 caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga
     7921 aaaggtggta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     7981 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     8041 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     8101 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     8161 gataataatg gtttcttagt atgatccaat atcaaaggaa atgatagcat tgaaggatga
     8221 gactaatcca attgaggagt ggcagcatat agaacagcta aagggtagtg ctgaaggaag
     8281 catacgatac cccgcatgga atgggataat atcacaggag gtactagact acctttcatc
     8341 ctacataaat agacgcatat aagtacgcat ttaagcataa acacgcacta tgccgttctt
     8401 ctcatgtata tatatataca ggcaacacgc agatataggt gcgacgtgaa cagtgagctg
     8461 tatgtgcgca gctcgcgttg cattttcgga agcgctcgtt ttcggaaacg ctttgaagtt
     8521 cctattccga agttcctatt ctctagaaag tataggaact tcagagcgct tttgaaaacc
     8581 aaaagcgctc tgaagacgca ctttcaaaaa accaaaaacg caccggactg taacgagcta
     8641 ctaaaatatt gcgaataccg cttccacaaa cattgctcaa aagtatctct ttgctatata
     8701 tctctgtgct atatccctat ataacctacc catccacctt tcgctccttg aacttgcatc
     8761 taaactcgac ctctacattt tttatgttta tctctagtat tactctttag acaaaaaaat
     8821 tgtagtaaga actattcata gagtgaatcg aaaacaatac gaaaatgtaa acatttccta
     8881 tacgtagtat atagagacaa aatagaagaa accgttcata attttctgac caatgaagaa
     8941 tcatcaacgc tatcactttc tgttcacaaa gtatgcgcaa tccacatcgg tatagaatat
     9001 aatcggggat gcctttatct tgaaaaaatg cacccgcagc ttcgctagta atcagtaaac
     9061 gcgggaagtg gagtcaggct ttttttatgg aagagaaaat agacaccaaa gtagccttct
     9121 tctaacctta acggacctac agtgcaaaaa gttatcaaga gactgcatta tagagcgcac
     9181 aaaggagaaa aaaagtaatc taagatgctt tgttagaaaa atagcgctct cgggatgcat
     9241 ttttgtagaa caaaaaagaa gtatagattc tttgttggta aaatagcgct ctcgcgttgc
     9301 atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt
     9361 tgcatttttg ttttacaaaa atgaagcaca gattcttcgt tggtaaaata gcgctttcgc
     9421 gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa ttagcgctct
     9481 cgcgttgcat ttttgttcta caaaatgaag cacagatgct tcgttcaggt ggcacttttc
     9541 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     9601 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     9661 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     9721 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     9781 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     9841 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     9901 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     9961 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
    10021 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
    10081 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
    10141 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
    10201 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
    10261 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
    10321 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
    10381 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
    10441 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
    10501 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
    10561 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
    10621 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
    10681 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
    10741 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
    10801 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
    10861 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
    10921 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
    10981 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
    11041 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
    11101 ccgaag
//
