LOCUS       sequence-193428-       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 ARS416d sequence 
            TAGTGCACTTACCCCACGTT in yeast chromosome 4..
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-ARS416d
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     351..369
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(407..429)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     529..548
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        572..787
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             788..1591
                     /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"
     rep_origin      complement(1722..2177)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1859..1880)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2071..2090
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2303..2325
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2317..2334
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2318..2334
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2344..2363
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2344..2362
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2381..2628)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(2644..2664)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2677..6780)
                     /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"
     promoter        complement(6790..7494)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     misc_RNA        complement(7580..7655)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(7882..7902)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7882..7900)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7921..7937)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7921..7937)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7934..7956)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7945..7961
                     /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)."
     promoter        complement(join(7969..7975,7976..7993,7994..7999))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8014..8035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8152..8169)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8323..8911)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8403..8422)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9082..9873,9874..9942))
                     /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     9705..9724
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9943..10047)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(10074..11106,1..310)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(11037..11084)
                     /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)."
ORIGIN
        1 cttccgaaaa tgcaacgcga gctgcgcaca tacagctcac tgttcacgtc gcacctatat
       61 ctgcgtgttg cctgtatata tatatacatg agaagaacgg catagtgcgt gtttatgctt
      121 aaatgcgtac ttatatgcgt ctatttatgt aggatgaaag gtagtctagt acctcctgtg
      181 atattatccc attccatgcg gggtatcgta tgcttccttc agcactaccc tttagctgtt
      241 ctatatgctg ccactcctca attggattag tctcatcctt caatgctatc atttcctttg
      301 atattggatc atactaagaa accattatta tcatgacatt aacctataaa aataggcgta
      361 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
      421 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
      481 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
      541 agattgtact gagagtgcac cataccacct tttcaattca tcattttttt tttattcttt
      601 tttttgattt cggtttcctt gaaatttttt tgattcggta atctccgaac agaaggaaga
      661 acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg ttgaagaaac
      721 atgaaattgc ccagtattct taacccaact gcacagaaca aaaacctgca ggaaacgaag
      781 ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta gtcctgttgc
      841 tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt
      901 tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa tttgtttact
      961 aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta agccgctaaa
     1021 ggcattatcc gccaagtaca attttttact cttcgaagac agaaaatttg ctgacattgg
     1081 taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat gggcagacat
     1141 tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc aggcggcaga
     1201 agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat gcaagggctc
     1261 cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg acaaagattt
     1321 tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt acgattggtt
     1381 gattatgaca cccggtgtgg gtttagatga caagggagac gcattgggtc aacagtatag
     1441 aaccgtggat gatgtggtct ctacaggatc tgacattatt attgttggaa gaggactatt
     1501 tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag gctgggaagc
     1561 atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa atgcatgtat
     1621 actaaactca caaattagag cttcaattta attatatcag ttattaccct atgcggtgtg
     1681 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
     1741 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     1801 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     1861 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     1921 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     1981 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2041 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2101 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2161 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     2221 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     2281 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
     2341 gcgtaatacg actcactata gggcgaattg ggtaccggcc gcaaattaaa gccttcgagc
     2401 gtcccaaaac cttctcaagc aaggttttca gtataatgtt acatgcgtac acgcgtctgt
     2461 acagaaaaaa aagaaaaatt tgaaatataa ataacgttct taatactaac ataactataa
     2521 aaaaataaat agggacctag acttcaggtt gtctaactcc ttccttttcg gttagagcgg
     2581 atgtgggggg agggcgtgaa cgtaagcgtg acataactaa ttacatgact cgagaagaga
     2641 tcacaccttc ctcttcttct tggggtcagc cctgctgtct ccaccgagct gagagaggtc
     2701 gattcttgtt tcatagagcc ccgtaattga ctgatgaatc agtgtggcgt ccaggacctc
     2761 ctttgtagag gtgtaccgct ttctgtctat ggtggtgtcg aagtacttga aggctgcagg
     2821 cgcgcccaag ttggtcagag taaacaagta gataatgttt tctgcctgct ccctgatggg
     2881 cttatccctg tgcttattgt aagcagaaag caccttatcg aggttagcgt cggcgaggat
     2941 cactcttttg gagaattcgc ttatttgctc gatgatctca tcaaggtagt gtttgtgttg
     3001 ttccacgaac agctgcttct gctcattatc ttcgggagac cctttgagct tttcatagtg
     3061 gctggccaga tacaagaaat taacgtattt agagggcagt gccagctcgt tacctttctg
     3121 cagctcgccc gcactagcga gcattcgttt ccggccgttt tcaagctcaa agagagagta
     3181 cttgggaagc ttaatgatga ggtctttttt gacctcttta tatcctttcg cctcgagaaa
     3241 gtcgatgggg tttttttcga agcttgatcg ctccatgatt gtgatgccca gcagttcctt
     3301 gacgcttttg agttttttag acttcccttt ctccactttg gccacaacca gtacactgta
     3361 agcgactgta ggagaatcga atccgccgta tttcttgggg tcccaatctt ttttgcgtgc
     3421 gatcagcttg tcgctgttcc ttttcgggag gatactttcc ttggagaagc ctccggtctg
     3481 tacttcggtc tttttaacga tgttcacctg cggcatggac aggaccttcc ggactgtcgc
     3541 gaaatcccta cccttgtccc acacgatttc tcctgtttct ccgtttgttt cgataagtgg
     3601 tcgcttccga atctctccat tggccagtgt aatctcggtc ttgaaaaaat tcataatatt
     3661 gctgtaaaag aagtacttag cggtggcctt gcctatttcc tgctcagact ttgcgatcat
     3721 tttcctaaca tcgtacactt tatagtctcc gtaaacaaat tcagattcaa gcttgggata
     3781 ttttttgata agtgcagtgc ctaccactgc attcaggtag gcatcatgcg catggtggta
     3841 attgttgatc tctctcacct tataaaactg aaagtccttt ctgaaatctg agaccagctt
     3901 agacttcaga gtaataactt tcacctctcg aatcagtttg tcattttcat cgtacttggt
     3961 gttcatgcgt gaatcgagaa tttgggccac gtgcttggtg atctggcgtg tctcaacaag
     4021 ctgccttttg atgaagccgg ctttatccaa ctcagacagg ccacctcgtt cagccttagt
     4081 cagattatcg aacttccgtt gtgtgatcag tttggcgttc agcagctgcc gccaataatt
     4141 tttcattttc ttgacaactt cttctgaggg gacgttatca ctcttccctc tatttttatc
     4201 ggatcttgtc aacactttat tatcaataga atcatctttg agaaaagact ggggcacgat
     4261 atgatccacg tcgtagtcgg agagccgatt gatgtccagt tcctgatcca cgtacatgtc
     4321 cctgccgttc tgcaggtagt acaggtagag cttctcattc tgaagctggg tgttttcaac
     4381 tgggtgttcc ttaaggattt gggaccccag ttcttttata ccctcttcaa tcctcttcat
     4441 cctttcccta ctgttcttct gtcccttctg ggtagtttgg ttctctcggg ccatctcgat
     4501 aacgatattc tcgggcttat gccttcccat tactttgacg agttcatcca cgaccttaac
     4561 ggtctgcagt attccctttt tgatagctgg gctacctgca agattagcga tgtgctcgtg
     4621 aagactgtcc ccctggccag aaacttgtgc tttctggatg tcctccttaa aggtgagaga
     4681 gtcatcatgg atcaactgca tgaagttccg gttggcaaat ccatcggact taagaaaatc
     4741 caggattgtc tttccactct gcttgtctcg gatcccattg atcagttttc ttgacagccg
     4801 cccccatcct gtatatcggc gcctcttgag ctgtttcatg actttgtcgt cgaagagatg
     4861 agcgtaagtt ttcaagcgtt cttcaatcat ctccctatct tcaaacaacg taagggtgag
     4921 gacaatgtcc tcaagaatgt cctcgttctc ctcattgtcc aggaagtcct tgtctttaat
     4981 gattttcagg agatcgtgat acgttcccag ggatgcgttg aagcgatcct ccactccgct
     5041 gatttcaaca gagtcgaaac attcaatctt tttgaaatag tcttctttga gctgtttcac
     5101 ggtaactttc cggttcgtct tgaagaggag gtccacgata gctttcttct gctctccaga
     5161 caggaatgct ggctttctca tcccttctgt gacgtatttg accttggtga gctcgttata
     5221 aactgtgaag tactcgtaca gcagagagtg tttaggaagc accttttcgt taggcagatt
     5281 tttatcaaag ttagtcatcc tttcgatgaa ggactgggca gaggccccct tatccacgac
     5341 ttcctcgaag ttccagggag tgatggtctc ttctgatttg cgagtcatcc acgcgaatct
     5401 ggaatttccc cgggcgaggg ggcctacata gtagggtatc cgaaatgtga ggattttctc
     5461 aatcttttcc ctgttatctt tcaaaaaggg gtagaaatcc tcttgccgcc tgaggatagc
     5521 gtgcagttcg cccaggtgaa tctggtgggg gatgcttcca ttgtcgaaag tgcgctgttt
     5581 gcgcaacaga tcttctctgt taagctttac cagcagctcc tcggtgccgt ccattttttc
     5641 caagatgggc ttaataaatt tgtaaaattc ctcctggctt gctccgccgt caatgtatcc
     5701 ggcgtagcca tttttagact gatcgaagaa aatttccttg tacttctcag gcagttgctg
     5761 tctgacaagg gccttcagca aagtcaagtc ttggtggtgc tcatcatagc gcttgatcat
     5821 actagcgctc agcggagctt tggtgatctc cgtgttcact cgcagaatat cactcagcag
     5881 aatggcgtct gacaggttct ttgccgccaa aaaaaggtct gcgtactggt cgccgatctg
     5941 ggccagcaga ttgtcgagat catcatcgta ggtgtctttg ctcagttgaa gcttggcatc
     6001 ttcggccagg tcgaagttag atttaaagtt gggggtcagc ccgagtgaca gggcgataag
     6061 attaccaaac aggccgttct tcttctcccc agggagctgt gcgatgaggt tttcgagccg
     6121 ccgggatttg gacagcctag cgctcaggat tgctttggcg tcaactccgg atgcgttgat
     6181 cgggttctct tcgaaaagct gattgtaagt ctgaaccagt tggataaaga gtttgtcgac
     6241 atcgctgttg tctgggttca ggtccccctc gatgaggaag tgtccccgaa atttgatcat
     6301 atgcgccagc gcgagataga tcaaccgcaa gtcagcctta tcagtactgt ctacaagctt
     6361 cttcctcaga tgatatatgg ttgggtactt ttcatggtac gccacctcgt ccacgatatt
     6421 gccaaagatt gggtggcgct cgttcttttt atcctcctcc accaaaaagg actcctccag
     6481 cctatggaag aaagagtcat ccaccttagc catctcatta ctaaagatct cctgcaggta
     6541 gcagatccga ttctttctgc gggtatatct gcgccgtgct gttcttttga gccgcgtggc
     6601 ttcggccgtc tccccggagt cgaacaggag ggcgccaatg aggttcttct ttatgctgtg
     6661 gcgatcggta ttgcccagaa ctttgaattt tttgctcggc accttgtact cgtccgtaat
     6721 gacggcccaa ccgacgctgt ttgtgccgat atcgagccca atggagtact tcttgtccat
     6781 ttttttccaa gttgattgta tgcttggtat agcttgaaat attgtgcaga aaaagaaaca
     6841 aggaagaaag ggaacgagaa caatgacgag gaaacaaaag attaataatt gcaggtctat
     6901 ttatacttga tagcaagaca gcaaactttt ttttatttca aattcaagta actggaagga
     6961 aggccgtata ccgttgctca ttagagagta gtgtgcgtga atgaaggaag gaaaaagttt
     7021 cgtgtgcttc gagatacccc tcatcagctc tggaacaacg acatctgttg gtgctgtctt
     7081 tgtcgttaat tttttccttt agtgtcttcc atcatttttt tgtcattgcg gatatggtga
     7141 gacaacaacg ggggagagag aaaagaaaaa aaaagaaaag aagttgcatg cgcctattat
     7201 tacttcaata gatggcaaat ggaaaaaggg tagtgaaact tcgatatgat gatggctatc
     7261 aagtctaggg ctacagtatt agttcgttat gtaccaccat caatgaggca gtgtaattgg
     7321 tgtagtcttg tttagcccat tatgtcttgt ctggtatctg ttctattgta tatctcccct
     7381 ccgccaccta catgttaggg agaccaacga aggtattata ggaatcccga tgtatgggtt
     7441 tggttgccag aaaagaggaa gtccatattg tacacccgga aacaacaaaa ggatgtcgac
     7501 tatccactag acagaagttt gcgttccata ctgtcagagg tggcatttta cataacaata
     7561 gtgacaaaaa ataaaaaaag caccgactcg gtgccacttt ttcaagttga taacggacta
     7621 gccttatttt aacttgctat ttctagctct aaaacaacgt ggggtaagtg cactaaaagt
     7681 cccattcgcc acccgaaggt gttgcccagc cggcgccagc gaggaggctg ggaccatgcc
     7741 ggccatctct cccgggggcg agtcgaacgc ccgatctcaa gatttcgtag tgataaatta
     7801 cagtcttgcg ccttaaacca acttggctac cgagagtatt taattgttga agaaagagta
     7861 tactagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     7921 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     7981 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     8041 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     8101 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     8161 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     8221 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     8281 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     8341 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     8401 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     8461 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     8521 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     8581 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     8641 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     8701 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     8761 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     8821 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     8881 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     8941 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     9001 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     9061 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     9121 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     9181 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     9241 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     9301 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     9361 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     9421 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     9481 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     9541 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     9601 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     9661 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     9721 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     9781 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     9841 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     9901 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9961 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
    10021 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaacgaag
    10081 catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
    10141 agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac
    10201 gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca
    10261 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta
    10321 ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt
    10381 ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc
    10441 ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat
    10501 tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc
    10561 tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt
    10621 gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta
    10681 tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt
    10741 attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat
    10801 actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa
    10861 ggtggatggg taggttatat agggatatag cacagagata tatagcaaag agatactttt
    10921 gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg
    10981 tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag cgctctgaag
    11041 ttcctatact ttctagagaa taggaacttc ggaataggaa cttcaaagcg tttccgaaaa
    11101 cgagcg
//
