LOCUS       sequence-193435-       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 ARS720a sequence 
            CAACAATTGTTACAATAGTA in yeast chromosome 7..
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-ARS720a
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(109..324)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(348..367)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     467..489
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(527..545)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      586..1928
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    918..965
                     /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        1955..2059
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2060..2128,2129..2920)
                     /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(2278..2297)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3091..3679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3580..3599
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3833..3850
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3967..3988
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4003..4008,4009..4026,4027..4033)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4041..4057
                     /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     4046..4068
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4065..4081
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4065..4081
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4100..4120
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4102..4120
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        4347..4422
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4508..5212
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             5222..9325
                     /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             9338..9358
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      9374..9621
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(9639..9658)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9640..9658)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9668..9685)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9668..9684)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9677..9699)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9825..10280
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9912..9931)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10122..10143
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(10411..11106,1..108))
                     /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 caatgaagca cacaagtttg tttgcttttc gtgcatgata ttaaatagct tggcagcaac
       61 aggactagga tgagtagcag cacgttcctt atatgtagct ttcgacatga tttatcttcg
      121 tttcctgcag gtttttgttc tgtgcagttg ggttaagaat actgggcaat ttcatgtttc
      181 ttcaacacta catatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc
      241 cttctgttcg gagattaccg aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga
      301 ataaaaaaaa aatgatgaat tgaaaaggtg gtatggtgca ctctcagtac aatctgctct
      361 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
      421 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
      481 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
      541 ctatttttat aggttaatgt catgataata atggtttctt agtatgatcc aatatcaaag
      601 gaaatgatag cattgaagga tgagactaat ccaattgagg agtggcagca tatagaacag
      661 ctaaagggta gtgctgaagg aagcatacga taccccgcat ggaatgggat aatatcacag
      721 gaggtactag actacctttc atcctacata aatagacgca tataagtacg catttaagca
      781 taaacacgca ctatgccgtt cttctcatgt atatatatat acaggcaaca cgcagatata
      841 ggtgcgacgt gaacagtgag ctgtatgtgc gcagctcgcg ttgcattttc ggaagcgctc
      901 gttttcggaa acgctttgaa gttcctattc cgaagttcct attctctaga aagtatagga
      961 acttcagagc gcttttgaaa accaaaagcg ctctgaagac gcactttcaa aaaaccaaaa
     1021 acgcaccgga ctgtaacgag ctactaaaat attgcgaata ccgcttccac aaacattgct
     1081 caaaagtatc tctttgctat atatctctgt gctatatccc tatataacct acccatccac
     1141 ctttcgctcc ttgaacttgc atctaaactc gacctctaca ttttttatgt ttatctctag
     1201 tattactctt tagacaaaaa aattgtagta agaactattc atagagtgaa tcgaaaacaa
     1261 tacgaaaatg taaacatttc ctatacgtag tatatagaga caaaatagaa gaaaccgttc
     1321 ataattttct gaccaatgaa gaatcatcaa cgctatcact ttctgttcac aaagtatgcg
     1381 caatccacat cggtatagaa tataatcggg gatgccttta tcttgaaaaa atgcacccgc
     1441 agcttcgcta gtaatcagta aacgcgggaa gtggagtcag gcttttttta tggaagagaa
     1501 aatagacacc aaagtagcct tcttctaacc ttaacggacc tacagtgcaa aaagttatca
     1561 agagactgca ttatagagcg cacaaaggag aaaaaaagta atctaagatg ctttgttaga
     1621 aaaatagcgc tctcgggatg catttttgta gaacaaaaaa gaagtataga ttctttgttg
     1681 gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa atgcagctca gattctttgt
     1741 ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca aaaatgaagc acagattctt
     1801 cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg taaaaatgca gctcagattc
     1861 tttgtttgaa aaattagcgc tctcgcgttg catttttgtt ctacaaaatg aagcacagat
     1921 gcttcgttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1981 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2041 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2101 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2161 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2221 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2281 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     2341 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     2401 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     2461 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     2521 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     2581 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     2641 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     2701 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     2761 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     2821 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2881 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     2941 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3001 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3061 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3121 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3181 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3241 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3301 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3361 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3421 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3481 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3541 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3601 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3661 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3721 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3781 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3841 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     3901 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     3961 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     4021 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     4081 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctagt
     4141 atactctttc ttcaacaatt aaatactctc ggtagccaag ttggtttaag gcgcaagact
     4201 gtaatttatc actacgaaat cttgagatcg ggcgttcgac tcgcccccgg gagagatggc
     4261 cggcatggtc ccagcctcct cgctggcgcc ggctgggcaa caccttcggg tggcgaatgg
     4321 gactttcaac aattgttaca atagtagttt tagagctaga aatagcaagt taaaataagg
     4381 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttta ttttttgtca
     4441 ctattgttat gtaaaatgcc acctctgaca gtatggaacg caaacttctg tctagtggat
     4501 agtcgacatc cttttgttgt ttccgggtgt acaatatgga cttcctcttt tctggcaacc
     4561 aaacccatac atcgggattc ctataatacc ttcgttggtc tccctaacat gtaggtggcg
     4621 gaggggagat atacaataga acagatacca gacaagacat aatgggctaa acaagactac
     4681 accaattaca ctgcctcatt gatggtggta cataacgaac taatactgta gccctagact
     4741 tgatagccat catcatatcg aagtttcact accctttttc catttgccat ctattgaagt
     4801 aataataggc gcatgcaact tcttttcttt ttttttcttt tctctctccc ccgttgttgt
     4861 ctcaccatat ccgcaatgac aaaaaaatga tggaagacac taaaggaaaa aattaacgac
     4921 aaagacagca ccaacagatg tcgttgttcc agagctgatg aggggtatct cgaagcacac
     4981 gaaacttttt ccttccttca ttcacgcaca ctactctcta atgagcaacg gtatacggcc
     5041 ttccttccag ttacttgaat ttgaaataaa aaaaagtttg ctgtcttgct atcaagtata
     5101 aatagacctg caattattaa tcttttgttt cctcgtcatt gttctcgttc cctttcttcc
     5161 ttgtttcttt ttctgcacaa tatttcaagc tataccaagc atacaatcaa cttggaaaaa
     5221 aatggacaag aagtactcca ttgggctcga tatcggcaca aacagcgtcg gttgggccgt
     5281 cattacggac gagtacaagg tgccgagcaa aaaattcaaa gttctgggca ataccgatcg
     5341 ccacagcata aagaagaacc tcattggcgc cctcctgttc gactccgggg agacggccga
     5401 agccacgcgg ctcaaaagaa cagcacggcg cagatatacc cgcagaaaga atcggatctg
     5461 ctacctgcag gagatcttta gtaatgagat ggctaaggtg gatgactctt tcttccatag
     5521 gctggaggag tcctttttgg tggaggagga taaaaagaac gagcgccacc caatctttgg
     5581 caatatcgtg gacgaggtgg cgtaccatga aaagtaccca accatatatc atctgaggaa
     5641 gaagcttgta gacagtactg ataaggctga cttgcggttg atctatctcg cgctggcgca
     5701 tatgatcaaa tttcggggac acttcctcat cgagggggac ctgaacccag acaacagcga
     5761 tgtcgacaaa ctctttatcc aactggttca gacttacaat cagcttttcg aagagaaccc
     5821 gatcaacgca tccggagttg acgccaaagc aatcctgagc gctaggctgt ccaaatcccg
     5881 gcggctcgaa aacctcatcg cacagctccc tggggagaag aagaacggcc tgtttggtaa
     5941 tcttatcgcc ctgtcactcg ggctgacccc caactttaaa tctaacttcg acctggccga
     6001 agatgccaag cttcaactga gcaaagacac ctacgatgat gatctcgaca atctgctggc
     6061 ccagatcggc gaccagtacg cagacctttt tttggcggca aagaacctgt cagacgccat
     6121 tctgctgagt gatattctgc gagtgaacac ggagatcacc aaagctccgc tgagcgctag
     6181 tatgatcaag cgctatgatg agcaccacca agacttgact ttgctgaagg cccttgtcag
     6241 acagcaactg cctgagaagt acaaggaaat tttcttcgat cagtctaaaa atggctacgc
     6301 cggatacatt gacggcggag caagccagga ggaattttac aaatttatta agcccatctt
     6361 ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt aacagagaag atctgttgcg
     6421 caaacagcgc actttcgaca atggaagcat cccccaccag attcacctgg gcgaactgca
     6481 cgctatcctc aggcggcaag aggatttcta cccctttttg aaagataaca gggaaaagat
     6541 tgagaaaatc ctcacatttc ggatacccta ctatgtaggc cccctcgccc ggggaaattc
     6601 cagattcgcg tggatgactc gcaaatcaga agagaccatc actccctgga acttcgagga
     6661 agtcgtggat aagggggcct ctgcccagtc cttcatcgaa aggatgacta actttgataa
     6721 aaatctgcct aacgaaaagg tgcttcctaa acactctctg ctgtacgagt acttcacagt
     6781 ttataacgag ctcaccaagg tcaaatacgt cacagaaggg atgagaaagc cagcattcct
     6841 gtctggagag cagaagaaag ctatcgtgga cctcctcttc aagacgaacc ggaaagttac
     6901 cgtgaaacag ctcaaagaag actatttcaa aaagattgaa tgtttcgact ctgttgaaat
     6961 cagcggagtg gaggatcgct tcaacgcatc cctgggaacg tatcacgatc tcctgaaaat
     7021 cattaaagac aaggacttcc tggacaatga ggagaacgag gacattcttg aggacattgt
     7081 cctcaccctt acgttgtttg aagataggga gatgattgaa gaacgcttga aaacttacgc
     7141 tcatctcttc gacgacaaag tcatgaaaca gctcaagagg cgccgatata caggatgggg
     7201 gcggctgtca agaaaactga tcaatgggat ccgagacaag cagagtggaa agacaatcct
     7261 ggattttctt aagtccgatg gatttgccaa ccggaacttc atgcagttga tccatgatga
     7321 ctctctcacc tttaaggagg acatccagaa agcacaagtt tctggccagg gggacagtct
     7381 tcacgagcac atcgctaatc ttgcaggtag cccagctatc aaaaagggaa tactgcagac
     7441 cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg cataagcccg agaatatcgt
     7501 tatcgagatg gcccgagaga accaaactac ccagaaggga cagaagaaca gtagggaaag
     7561 gatgaagagg attgaagagg gtataaaaga actggggtcc caaatcctta aggaacaccc
     7621 agttgaaaac acccagcttc agaatgagaa gctctacctg tactacctgc agaacggcag
     7681 ggacatgtac gtggatcagg aactggacat caatcggctc tccgactacg acgtggatca
     7741 tatcgtgccc cagtcttttc tcaaagatga ttctattgat aataaagtgt tgacaagatc
     7801 cgataaaaat agagggaaga gtgataacgt cccctcagaa gaagttgtca agaaaatgaa
     7861 aaattattgg cggcagctgc tgaacgccaa actgatcaca caacggaagt tcgataatct
     7921 gactaaggct gaacgaggtg gcctgtctga gttggataaa gccggcttca tcaaaaggca
     7981 gcttgttgag acacgccaga tcaccaagca cgtggcccaa attctcgatt cacgcatgaa
     8041 caccaagtac gatgaaaatg acaaactgat tcgagaggtg aaagttatta ctctgaagtc
     8101 taagctggtc tcagatttca gaaaggactt tcagttttat aaggtgagag agatcaacaa
     8161 ttaccaccat gcgcatgatg cctacctgaa tgcagtggta ggcactgcac ttatcaaaaa
     8221 atatcccaag cttgaatctg aatttgttta cggagactat aaagtgtacg atgttaggaa
     8281 aatgatcgca aagtctgagc aggaaatagg caaggccacc gctaagtact tcttttacag
     8341 caatattatg aattttttca agaccgagat tacactggcc aatggagaga ttcggaagcg
     8401 accacttatc gaaacaaacg gagaaacagg agaaatcgtg tgggacaagg gtagggattt
     8461 cgcgacagtc cggaaggtcc tgtccatgcc gcaggtgaac atcgttaaaa agaccgaagt
     8521 acagaccgga ggcttctcca aggaaagtat cctcccgaaa aggaacagcg acaagctgat
     8581 cgcacgcaaa aaagattggg accccaagaa atacggcgga ttcgattctc ctacagtcgc
     8641 ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag tctaaaaaac tcaaaagcgt
     8701 caaggaactg ctgggcatca caatcatgga gcgatcaagc ttcgaaaaaa accccatcga
     8761 ctttctcgag gcgaaaggat ataaagaggt caaaaaagac ctcatcatta agcttcccaa
     8821 gtactctctc tttgagcttg aaaacggccg gaaacgaatg ctcgctagtg cgggcgagct
     8881 gcagaaaggt aacgagctgg cactgccctc taaatacgtt aatttcttgt atctggccag
     8941 ccactatgaa aagctcaaag ggtctcccga agataatgag cagaagcagc tgttcgtgga
     9001 acaacacaaa cactaccttg atgagatcat cgagcaaata agcgaattct ccaaaagagt
     9061 gatcctcgcc gacgctaacc tcgataaggt gctttctgct tacaataagc acagggataa
     9121 gcccatcagg gagcaggcag aaaacattat ctacttgttt actctgacca acttgggcgc
     9181 gcctgcagcc ttcaagtact tcgacaccac catagacaga aagcggtaca cctctacaaa
     9241 ggaggtcctg gacgccacac tgattcatca gtcaattacg gggctctatg aaacaagaat
     9301 cgacctctct cagctcggtg gagacagcag ggctgacccc aagaagaaga ggaaggtgtg
     9361 atctcttctc gagtcatgta attagttatg tcacgcttac gttcacgccc tccccccaca
     9421 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     9481 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     9541 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
     9601 cgctcgaagg ctttaatttg cggccggtac ccaattcgcc ctatagtgag tcgtattacg
     9661 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     9721 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     9781 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg
     9841 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     9901 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     9961 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
    10021 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
    10081 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
    10141 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
    10201 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
    10261 aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
    10321 tcacaccgca tagggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag
    10381 tatacatgca tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata
    10441 tgcttcccag cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc
    10501 aaatagtcct cttccaacaa taataatgtc agatcctgta gagaccacat catccacggt
    10561 tctatactgt tgacccaatg cgtctccctt gtcatctaaa cccacaccgg gtgtcataat
    10621 caaccaatcg taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac
    10681 aaaatctttg tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag
    10741 ggagcccttg catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc
    10801 ttctgccgcc tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt
    10861 aatgtctgcc cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt
    10921 accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc
    10981 ctttagcggc ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt
    11041 tagtaaacaa attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg
    11101 aacatc
//
