LOCUS       sequence-193425-       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 ARS208a sequence 
            GTCCGCTAAACAAAAGATCT in yeast chromosome 2..
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-ARS208a
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(25..240)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(264..283)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     383..405
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(443..461)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      502..1844
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    834..881
                     /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        1871..1975
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1976..2044,2045..2836)
                     /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(2194..2213)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3007..3595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3496..3515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3749..3766
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3883..3904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3919..3924,3925..3942,3943..3949)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3957..3973
                     /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     3962..3984
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3981..3997
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3981..3997
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4016..4036
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4018..4036
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        4263..4338
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4424..5128
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             5138..9241
                     /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             9254..9274
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      9290..9537
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(9555..9574)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9556..9574)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9584..9601)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9584..9600)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9593..9615)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9741..10196
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9828..9847)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10038..10059
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(10327..11106,1..24))
                     /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 ttccttatat gtagctttcg acatgattta tcttcgtttc ctgcaggttt ttgttctgtg
       61 cagttgggtt aagaatactg ggcaatttca tgtttcttca acactacata tgcgtatata
      121 taccaatcta agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc
      181 aaaaaaattt caaggaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa
      241 aaggtggtat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
      301 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
      361 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
      421 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
      481 ataataatgg tttcttagta tgatccaata tcaaaggaaa tgatagcatt gaaggatgag
      541 actaatccaa ttgaggagtg gcagcatata gaacagctaa agggtagtgc tgaaggaagc
      601 atacgatacc ccgcatggaa tgggataata tcacaggagg tactagacta cctttcatcc
      661 tacataaata gacgcatata agtacgcatt taagcataaa cacgcactat gccgttcttc
      721 tcatgtatat atatatacag gcaacacgca gatataggtg cgacgtgaac agtgagctgt
      781 atgtgcgcag ctcgcgttgc attttcggaa gcgctcgttt tcggaaacgc tttgaagttc
      841 ctattccgaa gttcctattc tctagaaagt ataggaactt cagagcgctt ttgaaaacca
      901 aaagcgctct gaagacgcac tttcaaaaaa ccaaaaacgc accggactgt aacgagctac
      961 taaaatattg cgaataccgc ttccacaaac attgctcaaa agtatctctt tgctatatat
     1021 ctctgtgcta tatccctata taacctaccc atccaccttt cgctccttga acttgcatct
     1081 aaactcgacc tctacatttt ttatgtttat ctctagtatt actctttaga caaaaaaatt
     1141 gtagtaagaa ctattcatag agtgaatcga aaacaatacg aaaatgtaaa catttcctat
     1201 acgtagtata tagagacaaa atagaagaaa ccgttcataa ttttctgacc aatgaagaat
     1261 catcaacgct atcactttct gttcacaaag tatgcgcaat ccacatcggt atagaatata
     1321 atcggggatg cctttatctt gaaaaaatgc acccgcagct tcgctagtaa tcagtaaacg
     1381 cgggaagtgg agtcaggctt tttttatgga agagaaaata gacaccaaag tagccttctt
     1441 ctaaccttaa cggacctaca gtgcaaaaag ttatcaagag actgcattat agagcgcaca
     1501 aaggagaaaa aaagtaatct aagatgcttt gttagaaaaa tagcgctctc gggatgcatt
     1561 tttgtagaac aaaaaagaag tatagattct ttgttggtaa aatagcgctc tcgcgttgca
     1621 tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt
     1681 gcatttttgt tttacaaaaa tgaagcacag attcttcgtt ggtaaaatag cgctttcgcg
     1741 ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc
     1801 gcgttgcatt tttgttctac aaaatgaagc acagatgctt cgttcaggtg gcacttttcg
     1861 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1921 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1981 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2041 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2101 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2161 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2221 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2281 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2341 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2401 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2461 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2521 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2581 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2641 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2701 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2761 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2821 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2881 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2941 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3001 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3061 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3121 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3421 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3481 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3541 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3601 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3661 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3721 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3781 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     3841 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     3901 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     3961 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
     4021 taaccctcac taaagggaac aaaagctgga gctagtatac tctttcttca acaattaaat
     4081 actctcggta gccaagttgg tttaaggcgc aagactgtaa tttatcacta cgaaatcttg
     4141 agatcgggcg ttcgactcgc ccccgggaga gatggccggc atggtcccag cctcctcgct
     4201 ggcgccggct gggcaacacc ttcgggtggc gaatgggact ttgtccgcta aacaaaagat
     4261 ctgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
     4321 gtggcaccga gtcggtgctt tttttatttt ttgtcactat tgttatgtaa aatgccacct
     4381 ctgacagtat ggaacgcaaa cttctgtcta gtggatagtc gacatccttt tgttgtttcc
     4441 gggtgtacaa tatggacttc ctcttttctg gcaaccaaac ccatacatcg ggattcctat
     4501 aataccttcg ttggtctccc taacatgtag gtggcggagg ggagatatac aatagaacag
     4561 ataccagaca agacataatg ggctaaacaa gactacacca attacactgc ctcattgatg
     4621 gtggtacata acgaactaat actgtagccc tagacttgat agccatcatc atatcgaagt
     4681 ttcactaccc tttttccatt tgccatctat tgaagtaata ataggcgcat gcaacttctt
     4741 ttcttttttt ttcttttctc tctcccccgt tgttgtctca ccatatccgc aatgacaaaa
     4801 aaatgatgga agacactaaa ggaaaaaatt aacgacaaag acagcaccaa cagatgtcgt
     4861 tgttccagag ctgatgaggg gtatctcgaa gcacacgaaa ctttttcctt ccttcattca
     4921 cgcacactac tctctaatga gcaacggtat acggccttcc ttccagttac ttgaatttga
     4981 aataaaaaaa agtttgctgt cttgctatca agtataaata gacctgcaat tattaatctt
     5041 ttgtttcctc gtcattgttc tcgttccctt tcttccttgt ttctttttct gcacaatatt
     5101 tcaagctata ccaagcatac aatcaacttg gaaaaaaatg gacaagaagt actccattgg
     5161 gctcgatatc ggcacaaaca gcgtcggttg ggccgtcatt acggacgagt acaaggtgcc
     5221 gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac agcataaaga agaacctcat
     5281 tggcgccctc ctgttcgact ccggggagac ggccgaagcc acgcggctca aaagaacagc
     5341 acggcgcaga tatacccgca gaaagaatcg gatctgctac ctgcaggaga tctttagtaa
     5401 tgagatggct aaggtggatg actctttctt ccataggctg gaggagtcct ttttggtgga
     5461 ggaggataaa aagaacgagc gccacccaat ctttggcaat atcgtggacg aggtggcgta
     5521 ccatgaaaag tacccaacca tatatcatct gaggaagaag cttgtagaca gtactgataa
     5581 ggctgacttg cggttgatct atctcgcgct ggcgcatatg atcaaatttc ggggacactt
     5641 cctcatcgag ggggacctga acccagacaa cagcgatgtc gacaaactct ttatccaact
     5701 ggttcagact tacaatcagc ttttcgaaga gaacccgatc aacgcatccg gagttgacgc
     5761 caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca
     5821 gctccctggg gagaagaaga acggcctgtt tggtaatctt atcgccctgt cactcgggct
     5881 gacccccaac tttaaatcta acttcgacct ggccgaagat gccaagcttc aactgagcaa
     5941 agacacctac gatgatgatc tcgacaatct gctggcccag atcggcgacc agtacgcaga
     6001 cctttttttg gcggcaaaga acctgtcaga cgccattctg ctgagtgata ttctgcgagt
     6061 gaacacggag atcaccaaag ctccgctgag cgctagtatg atcaagcgct atgatgagca
     6121 ccaccaagac ttgactttgc tgaaggccct tgtcagacag caactgcctg agaagtacaa
     6181 ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga tacattgacg gcggagcaag
     6241 ccaggaggaa ttttacaaat ttattaagcc catcttggaa aaaatggacg gcaccgagga
     6301 gctgctggta aagcttaaca gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg
     6361 aagcatcccc caccagattc acctgggcga actgcacgct atcctcaggc ggcaagagga
     6421 tttctacccc tttttgaaag ataacaggga aaagattgag aaaatcctca catttcggat
     6481 accctactat gtaggccccc tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa
     6541 atcagaagag accatcactc cctggaactt cgaggaagtc gtggataagg gggcctctgc
     6601 ccagtccttc atcgaaagga tgactaactt tgataaaaat ctgcctaacg aaaaggtgct
     6661 tcctaaacac tctctgctgt acgagtactt cacagtttat aacgagctca ccaaggtcaa
     6721 atacgtcaca gaagggatga gaaagccagc attcctgtct ggagagcaga agaaagctat
     6781 cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg aaacagctca aagaagacta
     6841 tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa
     6901 cgcatccctg ggaacgtatc acgatctcct gaaaatcatt aaagacaagg acttcctgga
     6961 caatgaggag aacgaggaca ttcttgagga cattgtcctc acccttacgt tgtttgaaga
     7021 tagggagatg attgaagaac gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat
     7081 gaaacagctc aagaggcgcc gatatacagg atgggggcgg ctgtcaagaa aactgatcaa
     7141 tgggatccga gacaagcaga gtggaaagac aatcctggat tttcttaagt ccgatggatt
     7201 tgccaaccgg aacttcatgc agttgatcca tgatgactct ctcaccttta aggaggacat
     7261 ccagaaagca caagtttctg gccaggggga cagtcttcac gagcacatcg ctaatcttgc
     7321 aggtagccca gctatcaaaa agggaatact gcagaccgtt aaggtcgtgg atgaactcgt
     7381 caaagtaatg ggaaggcata agcccgagaa tatcgttatc gagatggccc gagagaacca
     7441 aactacccag aagggacaga agaacagtag ggaaaggatg aagaggattg aagagggtat
     7501 aaaagaactg gggtcccaaa tccttaagga acacccagtt gaaaacaccc agcttcagaa
     7561 tgagaagctc tacctgtact acctgcagaa cggcagggac atgtacgtgg atcaggaact
     7621 ggacatcaat cggctctccg actacgacgt ggatcatatc gtgccccagt cttttctcaa
     7681 agatgattct attgataata aagtgttgac aagatccgat aaaaatagag ggaagagtga
     7741 taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa
     7801 cgccaaactg atcacacaac ggaagttcga taatctgact aaggctgaac gaggtggcct
     7861 gtctgagttg gataaagccg gcttcatcaa aaggcagctt gttgagacac gccagatcac
     7921 caagcacgtg gcccaaattc tcgattcacg catgaacacc aagtacgatg aaaatgacaa
     7981 actgattcga gaggtgaaag ttattactct gaagtctaag ctggtctcag atttcagaaa
     8041 ggactttcag ttttataagg tgagagagat caacaattac caccatgcgc atgatgccta
     8101 cctgaatgca gtggtaggca ctgcacttat caaaaaatat cccaagcttg aatctgaatt
     8161 tgtttacgga gactataaag tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga
     8221 aataggcaag gccaccgcta agtacttctt ttacagcaat attatgaatt ttttcaagac
     8281 cgagattaca ctggccaatg gagagattcg gaagcgacca cttatcgaaa caaacggaga
     8341 aacaggagaa atcgtgtggg acaagggtag ggatttcgcg acagtccgga aggtcctgtc
     8401 catgccgcag gtgaacatcg ttaaaaagac cgaagtacag accggaggct tctccaagga
     8461 aagtatcctc ccgaaaagga acagcgacaa gctgatcgca cgcaaaaaag attgggaccc
     8521 caagaaatac ggcggattcg attctcctac agtcgcttac agtgtactgg ttgtggccaa
     8581 agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat
     8641 catggagcga tcaagcttcg aaaaaaaccc catcgacttt ctcgaggcga aaggatataa
     8701 agaggtcaaa aaagacctca tcattaagct tcccaagtac tctctctttg agcttgaaaa
     8761 cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag aaaggtaacg agctggcact
     8821 gccctctaaa tacgttaatt tcttgtatct ggccagccac tatgaaaagc tcaaagggtc
     8881 tcccgaagat aatgagcaga agcagctgtt cgtggaacaa cacaaacact accttgatga
     8941 gatcatcgag caaataagcg aattctccaa aagagtgatc ctcgccgacg ctaacctcga
     9001 taaggtgctt tctgcttaca ataagcacag ggataagccc atcagggagc aggcagaaaa
     9061 cattatctac ttgtttactc tgaccaactt gggcgcgcct gcagccttca agtacttcga
     9121 caccaccata gacagaaagc ggtacacctc tacaaaggag gtcctggacg ccacactgat
     9181 tcatcagtca attacggggc tctatgaaac aagaatcgac ctctctcagc tcggtggaga
     9241 cagcagggct gaccccaaga agaagaggaa ggtgtgatct cttctcgagt catgtaatta
     9301 gttatgtcac gcttacgttc acgccctccc cccacatccg ctctaaccga aaaggaagga
     9361 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
     9421 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
     9481 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc
     9541 cggtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     9601 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     9661 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     9721 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     9781 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     9841 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     9901 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     9961 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
    10021 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
    10081 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
    10141 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
    10201 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatagg gtaataactg
    10261 atataattaa attgaagctc taatttgtga gtttagtata catgcattta cttataatac
    10321 agttttttag ttttgctggc cgcatcttct caaatatgct tcccagcctg cttttctgta
    10381 acgttcaccc tctaccttag catcccttcc ctttgcaaat agtcctcttc caacaataat
    10441 aatgtcagat cctgtagaga ccacatcatc cacggttcta tactgttgac ccaatgcgtc
    10501 tcccttgtca tctaaaccca caccgggtgt cataatcaac caatcgtaac cttcatctct
    10561 tccacccatg tctctttgag caataaagcc gataacaaaa tctttgtcgc tcttcgcaat
    10621 gtcaacagta cccttagtat attctccagt agatagggag cccttgcatg acaattctgc
    10681 taacatcaaa aggcctctag gttcctttgt tacttcttct gccgcctgct tcaaaccgct
    10741 aacaatacct gggcccacca caccgtgtgc attcgtaatg tctgcccatt ctgctattct
    10801 gtatacaccc gcagagtact gcaatttgac tgtattacca atgtcagcaa attttctgtc
    10861 ttcgaagagt aaaaaattgt acttggcgga taatgccttt agcggcttaa ctgtgccctc
    10921 catggaaaaa tcagtcaaga tatccacatg tgtttttagt aaacaaattt tgggacctaa
    10981 tgcttcaact aactccagta attccttggt ggtacgaaca tccaatgaag cacacaagtt
    11041 tgtttgcttt tcgtgcatga tattaaatag cttggcagca acaggactag gatgagtagc
    11101 agcacg
//
