LOCUS       sequence-193426-       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 ARS308a sequence 
            CACTTGTCAAACAGAATATA in yeast chromosome 3.
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-ARS308a
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(69..284)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(308..327)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     427..449
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(487..505)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      546..1888
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    878..925
                     /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        1915..2019
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2020..2088,2089..2880)
                     /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(2238..2257)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3051..3639
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3540..3559
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3793..3810
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3927..3948
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3963..3968,3969..3986,3987..3993)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4001..4017
                     /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     4006..4028
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4025..4041
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4025..4041
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4060..4080
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4062..4080
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        4307..4382
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4468..5172
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             5182..9285
                     /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             9298..9318
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      9334..9581
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(9599..9618)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9600..9618)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9628..9645)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9628..9644)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9637..9659)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9785..10240
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9872..9891)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10082..10103
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(10371..11106,1..68))
                     /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"
ORIGIN
        1 ttaaatagct tggcagcaac aggactagga tgagtagcag cacgttcctt atatgtagct
       61 ttcgacatga tttatcttcg tttcctgcag gtttttgttc tgtgcagttg ggttaagaat
      121 actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa tctaagtctg
      181 tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa atttcaagga
      241 aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaaggtg gtatggtgca
      301 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
      361 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
      421 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
      481 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
      541 agtatgatcc aatatcaaag gaaatgatag cattgaagga tgagactaat ccaattgagg
      601 agtggcagca tatagaacag ctaaagggta gtgctgaagg aagcatacga taccccgcat
      661 ggaatgggat aatatcacag gaggtactag actacctttc atcctacata aatagacgca
      721 tataagtacg catttaagca taaacacgca ctatgccgtt cttctcatgt atatatatat
      781 acaggcaaca cgcagatata ggtgcgacgt gaacagtgag ctgtatgtgc gcagctcgcg
      841 ttgcattttc ggaagcgctc gttttcggaa acgctttgaa gttcctattc cgaagttcct
      901 attctctaga aagtatagga acttcagagc gcttttgaaa accaaaagcg ctctgaagac
      961 gcactttcaa aaaaccaaaa acgcaccgga ctgtaacgag ctactaaaat attgcgaata
     1021 ccgcttccac aaacattgct caaaagtatc tctttgctat atatctctgt gctatatccc
     1081 tatataacct acccatccac ctttcgctcc ttgaacttgc atctaaactc gacctctaca
     1141 ttttttatgt ttatctctag tattactctt tagacaaaaa aattgtagta agaactattc
     1201 atagagtgaa tcgaaaacaa tacgaaaatg taaacatttc ctatacgtag tatatagaga
     1261 caaaatagaa gaaaccgttc ataattttct gaccaatgaa gaatcatcaa cgctatcact
     1321 ttctgttcac aaagtatgcg caatccacat cggtatagaa tataatcggg gatgccttta
     1381 tcttgaaaaa atgcacccgc agcttcgcta gtaatcagta aacgcgggaa gtggagtcag
     1441 gcttttttta tggaagagaa aatagacacc aaagtagcct tcttctaacc ttaacggacc
     1501 tacagtgcaa aaagttatca agagactgca ttatagagcg cacaaaggag aaaaaaagta
     1561 atctaagatg ctttgttaga aaaatagcgc tctcgggatg catttttgta gaacaaaaaa
     1621 gaagtataga ttctttgttg gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa
     1681 atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca
     1741 aaaatgaagc acagattctt cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg
     1801 taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt
     1861 ctacaaaatg aagcacagat gcttcgttca ggtggcactt ttcggggaaa tgtgcgcgga
     1921 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1981 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2041 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2101 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2161 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2221 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2281 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2341 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2401 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2461 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2521 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2581 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2641 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2701 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2761 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2821 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2881 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2941 aaaaggatct aggtgaagat cctttttgat aatctcatga acaaaatccc ttaacgtgag
     3001 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3061 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3121 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3181 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3241 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3301 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3361 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3421 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3481 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3541 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3601 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3661 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3721 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3781 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3841 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3901 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     3961 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     4021 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     4081 gaacaaaagc tggagctagt atactctttc ttcaacaatt aaatactctc ggtagccaag
     4141 ttggtttaag gcgcaagact gtaatttatc actacgaaat cttgagatcg ggcgttcgac
     4201 tcgcccccgg gagagatggc cggcatggtc ccagcctcct cgctggcgcc ggctgggcaa
     4261 caccttcggg tggcgaatgg gactttcact tgtcaaacag aatatagttt tagagctaga
     4321 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
     4381 gcttttttta ttttttgtca ctattgttat gtaaaatgcc acctctgaca gtatggaacg
     4441 caaacttctg tctagtggat agtcgacatc cttttgttgt ttccgggtgt acaatatgga
     4501 cttcctcttt tctggcaacc aaacccatac atcgggattc ctataatacc ttcgttggtc
     4561 tccctaacat gtaggtggcg gaggggagat atacaataga acagatacca gacaagacat
     4621 aatgggctaa acaagactac accaattaca ctgcctcatt gatggtggta cataacgaac
     4681 taatactgta gccctagact tgatagccat catcatatcg aagtttcact accctttttc
     4741 catttgccat ctattgaagt aataataggc gcatgcaact tcttttcttt ttttttcttt
     4801 tctctctccc ccgttgttgt ctcaccatat ccgcaatgac aaaaaaatga tggaagacac
     4861 taaaggaaaa aattaacgac aaagacagca ccaacagatg tcgttgttcc agagctgatg
     4921 aggggtatct cgaagcacac gaaacttttt ccttccttca ttcacgcaca ctactctcta
     4981 atgagcaacg gtatacggcc ttccttccag ttacttgaat ttgaaataaa aaaaagtttg
     5041 ctgtcttgct atcaagtata aatagacctg caattattaa tcttttgttt cctcgtcatt
     5101 gttctcgttc cctttcttcc ttgtttcttt ttctgcacaa tatttcaagc tataccaagc
     5161 atacaatcaa cttggaaaaa aatggacaag aagtactcca ttgggctcga tatcggcaca
     5221 aacagcgtcg gttgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa
     5281 gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggcgc cctcctgttc
     5341 gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc
     5401 cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat ggctaaggtg
     5461 gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagaac
     5521 gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca
     5581 accatatatc atctgaggaa gaagcttgta gacagtactg ataaggctga cttgcggttg
     5641 atctatctcg cgctggcgca tatgatcaaa tttcggggac acttcctcat cgagggggac
     5701 ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat
     5761 cagcttttcg aagagaaccc gatcaacgca tccggagttg acgccaaagc aatcctgagc
     5821 gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag
     5881 aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa
     5941 tctaacttcg acctggccga agatgccaag cttcaactga gcaaagacac ctacgatgat
     6001 gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca
     6061 aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc
     6121 aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact
     6181 ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat
     6241 cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga ggaattttac
     6301 aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt
     6361 aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag
     6421 attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg
     6481 aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc
     6541 cccctcgccc ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc
     6601 actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa
     6661 aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg
     6721 ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg
     6781 atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc
     6841 aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa aaagattgaa
     6901 tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg
     6961 tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag
     7021 gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa
     7081 gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagagg
     7141 cgccgatata caggatgggg gcggctgtca agaaaactga tcaatgggat ccgagacaag
     7201 cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa ccggaacttc
     7261 atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa agcacaagtt
     7321 tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag cccagctatc
     7381 aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg
     7441 cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac ccagaaggga
     7501 cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga actggggtcc
     7561 caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa gctctacctg
     7621 tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat caatcggctc
     7681 tccgactacg acgtggatca tatcgtgccc cagtcttttc tcaaagatga ttctattgat
     7741 aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt cccctcagaa
     7801 gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa actgatcaca
     7861 caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga gttggataaa
     7921 gccggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca cgtggcccaa
     7981 attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat tcgagaggtg
     8041 aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt tcagttttat
     8101 aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa tgcagtggta
     8161 ggcactgcac ttatcaaaaa atatcccaag cttgaatctg aatttgttta cggagactat
     8221 aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg caaggccacc
     8281 gctaagtact tcttttacag caatattatg aattttttca agaccgagat tacactggcc
     8341 aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg agaaatcgtg
     8401 tgggacaagg gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc gcaggtgaac
     8461 atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat cctcccgaaa
     8521 aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa atacggcgga
     8581 ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag
     8641 tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caatcatgga gcgatcaagc
     8701 ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat ataaagaggt caaaaaagac
     8761 ctcatcatta agcttcccaa gtactctctc tttgagcttg aaaacggccg gaaacgaatg
     8821 ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc taaatacgtt
     8881 aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga agataatgag
     8941 cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat cgagcaaata
     9001 agcgaattct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt gctttctgct
     9061 tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat ctacttgttt
     9121 actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac catagacaga
     9181 aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca gtcaattacg
     9241 gggctctatg aaacaagaat cgacctctct cagctcggtg gagacagcag ggctgacccc
     9301 aagaagaaga ggaaggtgtg atctcttctc gagtcatgta attagttatg tcacgcttac
     9361 gttcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
     9421 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     9481 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     9541 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cggccggtac ccaattcgcc
     9601 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     9661 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     9721 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     9781 atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     9841 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     9901 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     9961 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
    10021 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
    10081 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
    10141 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
    10201 tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
    10261 ttacgcatct gtgcggtatt tcacaccgca tagggtaata actgatataa ttaaattgaa
    10321 gctctaattt gtgagtttag tatacatgca tttacttata atacagtttt ttagttttgc
    10381 tggccgcatc ttctcaaata tgcttcccag cctgcttttc tgtaacgttc accctctacc
    10441 ttagcatccc ttccctttgc aaatagtcct cttccaacaa taataatgtc agatcctgta
    10501 gagaccacat catccacggt tctatactgt tgacccaatg cgtctccctt gtcatctaaa
    10561 cccacaccgg gtgtcataat caaccaatcg taaccttcat ctcttccacc catgtctctt
    10621 tgagcaataa agccgataac aaaatctttg tcgctcttcg caatgtcaac agtaccctta
    10681 gtatattctc cagtagatag ggagcccttg catgacaatt ctgctaacat caaaaggcct
    10741 ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac cgctaacaat acctgggccc
    10801 accacaccgt gtgcattcgt aatgtctgcc cattctgcta ttctgtatac acccgcagag
    10861 tactgcaatt tgactgtatt accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa
    10921 ttgtacttgg cggataatgc ctttagcggc ttaactgtgc cctccatgga aaaatcagtc
    10981 aagatatcca catgtgtttt tagtaaacaa attttgggac ctaatgcttc aactaactcc
    11041 agtaattcct tggtggtacg aacatccaat gaagcacaca agtttgtttg cttttcgtgc
    11101 atgata
//
