LOCUS       sequence-193438-       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 ARS1021b sequence 
            CCTCTGTGTGGTGGTAATTG 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-ARS1021b
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(48..263)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(287..306)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     406..428
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(466..484)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      525..1867
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    857..904
                     /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        1894..1998
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1999..2067,2068..2859)
                     /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(2217..2236)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3030..3618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3519..3538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3772..3789
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3906..3927
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3942..3947,3948..3965,3966..3972)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3980..3996
                     /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     3985..4007
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4004..4020
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4004..4020
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4039..4059
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4041..4059
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        4286..4361
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4447..5151
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             5161..9264
                     /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             9277..9297
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      9313..9560
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(9578..9597)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9579..9597)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9607..9624)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9607..9623)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9616..9638)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9764..10219
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9851..9870)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10061..10082
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(10350..11106,1..47))
                     /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 ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt
       61 ttcctgcagg tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct
      121 tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc
      181 ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa
      241 taaaaaaaaa atgatgaatt gaaaaggtgg tatggtgcac tctcagtaca atctgctctg
      301 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
      361 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
      421 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
      481 tatttttata ggttaatgtc atgataataa tggtttctta gtatgatcca atatcaaagg
      541 aaatgatagc attgaaggat gagactaatc caattgagga gtggcagcat atagaacagc
      601 taaagggtag tgctgaagga agcatacgat accccgcatg gaatgggata atatcacagg
      661 aggtactaga ctacctttca tcctacataa atagacgcat ataagtacgc atttaagcat
      721 aaacacgcac tatgccgttc ttctcatgta tatatatata caggcaacac gcagatatag
      781 gtgcgacgtg aacagtgagc tgtatgtgcg cagctcgcgt tgcattttcg gaagcgctcg
      841 ttttcggaaa cgctttgaag ttcctattcc gaagttccta ttctctagaa agtataggaa
      901 cttcagagcg cttttgaaaa ccaaaagcgc tctgaagacg cactttcaaa aaaccaaaaa
      961 cgcaccggac tgtaacgagc tactaaaata ttgcgaatac cgcttccaca aacattgctc
     1021 aaaagtatct ctttgctata tatctctgtg ctatatccct atataaccta cccatccacc
     1081 tttcgctcct tgaacttgca tctaaactcg acctctacat tttttatgtt tatctctagt
     1141 attactcttt agacaaaaaa attgtagtaa gaactattca tagagtgaat cgaaaacaat
     1201 acgaaaatgt aaacatttcc tatacgtagt atatagagac aaaatagaag aaaccgttca
     1261 taattttctg accaatgaag aatcatcaac gctatcactt tctgttcaca aagtatgcgc
     1321 aatccacatc ggtatagaat ataatcgggg atgcctttat cttgaaaaaa tgcacccgca
     1381 gcttcgctag taatcagtaa acgcgggaag tggagtcagg ctttttttat ggaagagaaa
     1441 atagacacca aagtagcctt cttctaacct taacggacct acagtgcaaa aagttatcaa
     1501 gagactgcat tatagagcgc acaaaggaga aaaaaagtaa tctaagatgc tttgttagaa
     1561 aaatagcgct ctcgggatgc atttttgtag aacaaaaaag aagtatagat tctttgttgg
     1621 taaaatagcg ctctcgcgtt gcatttctgt tctgtaaaaa tgcagctcag attctttgtt
     1681 tgaaaaatta gcgctctcgc gttgcatttt tgttttacaa aaatgaagca cagattcttc
     1741 gttggtaaaa tagcgctttc gcgttgcatt tctgttctgt aaaaatgcag ctcagattct
     1801 ttgtttgaaa aattagcgct ctcgcgttgc atttttgttc tacaaaatga agcacagatg
     1861 cttcgttcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1921 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1981 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2041 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2101 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2161 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     2221 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     2281 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     2341 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2401 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2461 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2521 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2581 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2641 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2701 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2761 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2821 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2881 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2941 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3001 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3061 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3121 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3181 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3241 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3301 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3361 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3421 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3481 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3541 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3601 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3661 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3721 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3781 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     3841 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3901 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     3961 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     4021 catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctagta
     4081 tactctttct tcaacaatta aatactctcg gtagccaagt tggtttaagg cgcaagactg
     4141 taatttatca ctacgaaatc ttgagatcgg gcgttcgact cgcccccggg agagatggcc
     4201 ggcatggtcc cagcctcctc gctggcgccg gctgggcaac accttcgggt ggcgaatggg
     4261 actttcctct gtgtggtggt aattggtttt agagctagaa atagcaagtt aaaataaggc
     4321 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttat tttttgtcac
     4381 tattgttatg taaaatgcca cctctgacag tatggaacgc aaacttctgt ctagtggata
     4441 gtcgacatcc ttttgttgtt tccgggtgta caatatggac ttcctctttt ctggcaacca
     4501 aacccataca tcgggattcc tataatacct tcgttggtct ccctaacatg taggtggcgg
     4561 aggggagata tacaatagaa cagataccag acaagacata atgggctaaa caagactaca
     4621 ccaattacac tgcctcattg atggtggtac ataacgaact aatactgtag ccctagactt
     4681 gatagccatc atcatatcga agtttcacta ccctttttcc atttgccatc tattgaagta
     4741 ataataggcg catgcaactt cttttctttt tttttctttt ctctctcccc cgttgttgtc
     4801 tcaccatatc cgcaatgaca aaaaaatgat ggaagacact aaaggaaaaa attaacgaca
     4861 aagacagcac caacagatgt cgttgttcca gagctgatga ggggtatctc gaagcacacg
     4921 aaactttttc cttccttcat tcacgcacac tactctctaa tgagcaacgg tatacggcct
     4981 tccttccagt tacttgaatt tgaaataaaa aaaagtttgc tgtcttgcta tcaagtataa
     5041 atagacctgc aattattaat cttttgtttc ctcgtcattg ttctcgttcc ctttcttcct
     5101 tgtttctttt tctgcacaat atttcaagct ataccaagca tacaatcaac ttggaaaaaa
     5161 atggacaaga agtactccat tgggctcgat atcggcacaa acagcgtcgg ttgggccgtc
     5221 attacggacg agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa taccgatcgc
     5281 cacagcataa agaagaacct cattggcgcc ctcctgttcg actccgggga gacggccgaa
     5341 gccacgcggc tcaaaagaac agcacggcgc agatataccc gcagaaagaa tcggatctgc
     5401 tacctgcagg agatctttag taatgagatg gctaaggtgg atgactcttt cttccatagg
     5461 ctggaggagt cctttttggt ggaggaggat aaaaagaacg agcgccaccc aatctttggc
     5521 aatatcgtgg acgaggtggc gtaccatgaa aagtacccaa ccatatatca tctgaggaag
     5581 aagcttgtag acagtactga taaggctgac ttgcggttga tctatctcgc gctggcgcat
     5641 atgatcaaat ttcggggaca cttcctcatc gagggggacc tgaacccaga caacagcgat
     5701 gtcgacaaac tctttatcca actggttcag acttacaatc agcttttcga agagaacccg
     5761 atcaacgcat ccggagttga cgccaaagca atcctgagcg ctaggctgtc caaatcccgg
     5821 cggctcgaaa acctcatcgc acagctccct ggggagaaga agaacggcct gtttggtaat
     5881 cttatcgccc tgtcactcgg gctgaccccc aactttaaat ctaacttcga cctggccgaa
     5941 gatgccaagc ttcaactgag caaagacacc tacgatgatg atctcgacaa tctgctggcc
     6001 cagatcggcg accagtacgc agaccttttt ttggcggcaa agaacctgtc agacgccatt
     6061 ctgctgagtg atattctgcg agtgaacacg gagatcacca aagctccgct gagcgctagt
     6121 atgatcaagc gctatgatga gcaccaccaa gacttgactt tgctgaaggc ccttgtcaga
     6181 cagcaactgc ctgagaagta caaggaaatt ttcttcgatc agtctaaaaa tggctacgcc
     6241 ggatacattg acggcggagc aagccaggag gaattttaca aatttattaa gcccatcttg
     6301 gaaaaaatgg acggcaccga ggagctgctg gtaaagctta acagagaaga tctgttgcgc
     6361 aaacagcgca ctttcgacaa tggaagcatc ccccaccaga ttcacctggg cgaactgcac
     6421 gctatcctca ggcggcaaga ggatttctac ccctttttga aagataacag ggaaaagatt
     6481 gagaaaatcc tcacatttcg gataccctac tatgtaggcc ccctcgcccg gggaaattcc
     6541 agattcgcgt ggatgactcg caaatcagaa gagaccatca ctccctggaa cttcgaggaa
     6601 gtcgtggata agggggcctc tgcccagtcc ttcatcgaaa ggatgactaa ctttgataaa
     6661 aatctgccta acgaaaaggt gcttcctaaa cactctctgc tgtacgagta cttcacagtt
     6721 tataacgagc tcaccaaggt caaatacgtc acagaaggga tgagaaagcc agcattcctg
     6781 tctggagagc agaagaaagc tatcgtggac ctcctcttca agacgaaccg gaaagttacc
     6841 gtgaaacagc tcaaagaaga ctatttcaaa aagattgaat gtttcgactc tgttgaaatc
     6901 agcggagtgg aggatcgctt caacgcatcc ctgggaacgt atcacgatct cctgaaaatc
     6961 attaaagaca aggacttcct ggacaatgag gagaacgagg acattcttga ggacattgtc
     7021 ctcaccctta cgttgtttga agatagggag atgattgaag aacgcttgaa aacttacgct
     7081 catctcttcg acgacaaagt catgaaacag ctcaagaggc gccgatatac aggatggggg
     7141 cggctgtcaa gaaaactgat caatgggatc cgagacaagc agagtggaaa gacaatcctg
     7201 gattttctta agtccgatgg atttgccaac cggaacttca tgcagttgat ccatgatgac
     7261 tctctcacct ttaaggagga catccagaaa gcacaagttt ctggccaggg ggacagtctt
     7321 cacgagcaca tcgctaatct tgcaggtagc ccagctatca aaaagggaat actgcagacc
     7381 gttaaggtcg tggatgaact cgtcaaagta atgggaaggc ataagcccga gaatatcgtt
     7441 atcgagatgg cccgagagaa ccaaactacc cagaagggac agaagaacag tagggaaagg
     7501 atgaagagga ttgaagaggg tataaaagaa ctggggtccc aaatccttaa ggaacaccca
     7561 gttgaaaaca cccagcttca gaatgagaag ctctacctgt actacctgca gaacggcagg
     7621 gacatgtacg tggatcagga actggacatc aatcggctct ccgactacga cgtggatcat
     7681 atcgtgcccc agtcttttct caaagatgat tctattgata ataaagtgtt gacaagatcc
     7741 gataaaaata gagggaagag tgataacgtc ccctcagaag aagttgtcaa gaaaatgaaa
     7801 aattattggc ggcagctgct gaacgccaaa ctgatcacac aacggaagtt cgataatctg
     7861 actaaggctg aacgaggtgg cctgtctgag ttggataaag ccggcttcat caaaaggcag
     7921 cttgttgaga cacgccagat caccaagcac gtggcccaaa ttctcgattc acgcatgaac
     7981 accaagtacg atgaaaatga caaactgatt cgagaggtga aagttattac tctgaagtct
     8041 aagctggtct cagatttcag aaaggacttt cagttttata aggtgagaga gatcaacaat
     8101 taccaccatg cgcatgatgc ctacctgaat gcagtggtag gcactgcact tatcaaaaaa
     8161 tatcccaagc ttgaatctga atttgtttac ggagactata aagtgtacga tgttaggaaa
     8221 atgatcgcaa agtctgagca ggaaataggc aaggccaccg ctaagtactt cttttacagc
     8281 aatattatga attttttcaa gaccgagatt acactggcca atggagagat tcggaagcga
     8341 ccacttatcg aaacaaacgg agaaacagga gaaatcgtgt gggacaaggg tagggatttc
     8401 gcgacagtcc ggaaggtcct gtccatgccg caggtgaaca tcgttaaaaa gaccgaagta
     8461 cagaccggag gcttctccaa ggaaagtatc ctcccgaaaa ggaacagcga caagctgatc
     8521 gcacgcaaaa aagattggga ccccaagaaa tacggcggat tcgattctcc tacagtcgct
     8581 tacagtgtac tggttgtggc caaagtggag aaagggaagt ctaaaaaact caaaagcgtc
     8641 aaggaactgc tgggcatcac aatcatggag cgatcaagct tcgaaaaaaa ccccatcgac
     8701 tttctcgagg cgaaaggata taaagaggtc aaaaaagacc tcatcattaa gcttcccaag
     8761 tactctctct ttgagcttga aaacggccgg aaacgaatgc tcgctagtgc gggcgagctg
     8821 cagaaaggta acgagctggc actgccctct aaatacgtta atttcttgta tctggccagc
     8881 cactatgaaa agctcaaagg gtctcccgaa gataatgagc agaagcagct gttcgtggaa
     8941 caacacaaac actaccttga tgagatcatc gagcaaataa gcgaattctc caaaagagtg
     9001 atcctcgccg acgctaacct cgataaggtg ctttctgctt acaataagca cagggataag
     9061 cccatcaggg agcaggcaga aaacattatc tacttgttta ctctgaccaa cttgggcgcg
     9121 cctgcagcct tcaagtactt cgacaccacc atagacagaa agcggtacac ctctacaaag
     9181 gaggtcctgg acgccacact gattcatcag tcaattacgg ggctctatga aacaagaatc
     9241 gacctctctc agctcggtgg agacagcagg gctgacccca agaagaagag gaaggtgtga
     9301 tctcttctcg agtcatgtaa ttagttatgt cacgcttacg ttcacgccct ccccccacat
     9361 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     9421 ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
     9481 acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac
     9541 gctcgaaggc tttaatttgc ggccggtacc caattcgccc tatagtgagt cgtattacgc
     9601 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     9661 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     9721 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc
     9781 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     9841 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     9901 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     9961 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
    10021 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
    10081 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
    10141 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
    10201 atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
    10261 cacaccgcat agggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt
    10321 atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat
    10381 gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca
    10441 aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt
    10501 ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc
    10561 aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca
    10621 aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg
    10681 gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct
    10741 tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta
    10801 atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta
    10861 ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc
    10921 tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt
    10981 agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga
    11041 acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca
    11101 gcaaca
//
