LOCUS       sequence-193431-       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 SAP155b sequence 
            GGTTTTCATACTGGGGCCGC in yeast chromosome 6.
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-SAP155b
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..52)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(206..794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(286..305)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(965..1756,1757..1825))
                     /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     1588..1607
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1826..1930)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1957..3299
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(2920..2967)
                     /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)."
     primer_bind     3340..3358
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3396..3418)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3518..3537
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3561..3776
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3777..4580
                     /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(4711..5166)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4848..4869)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5060..5079
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5292..5314
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5306..5323
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5307..5323
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5333..5352
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5333..5351
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(5370..5617)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(5633..5653)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(5666..9769)
                     /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(9779..10483)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     misc_RNA        complement(10569..10644)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(10871..10891)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(10871..10889)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10910..10926)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10910..10926)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10923..10945)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10934..10950
                     /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(10958..10964,10965..10982,10983..10988))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11003..11024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
       61 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
      961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1921 gggttccgcg cacatttccc cgaaaagtgc cacctgaacg aagcatctgt gcttcatttt
     1981 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     2041 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     2101 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     2161 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     2221 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     2281 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     2341 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     2401 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     2461 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     2521 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
     2581 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
     2641 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
     2701 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
     2761 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
     2821 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa
     2881 gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctaga
     2941 gaataggaac ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat
     3001 gcaacgcgag ctgcgcacat acagctcact gttcacgtcg cacctatatc tgcgtgttgc
     3061 ctgtatatat atatacatga gaagaacggc atagtgcgtg tttatgctta aatgcgtact
     3121 tatatgcgtc tatttatgta ggatgaaagg tagtctagta cctcctgtga tattatccca
     3181 ttccatgcgg ggtatcgtat gcttccttca gcactaccct ttagctgttc tatatgctgc
     3241 cactcctcaa ttggattagt ctcatccttc aatgctatca tttcctttga tattggatca
     3301 tactaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     3361 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     3421 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     3481 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     3541 agagtgcacc ataccacctt ttcaattcat catttttttt ttattctttt ttttgatttc
     3601 ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
     3661 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
     3721 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
     3781 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
     3841 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
     3901 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
     3961 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
     4021 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
     4081 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
     4141 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
     4201 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
     4261 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
     4321 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     4381 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     4441 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     4501 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     4561 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     4621 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     4681 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat
     4741 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4801 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4861 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4921 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4981 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     5041 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     5101 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     5161 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     5221 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     5281 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
     5341 ctcactatag ggcgaattgg gtaccggccg caaattaaag ccttcgagcg tcccaaaacc
     5401 ttctcaagca aggttttcag tataatgtta catgcgtaca cgcgtctgta cagaaaaaaa
     5461 agaaaaattt gaaatataaa taacgttctt aatactaaca taactataaa aaaataaata
     5521 gggacctaga cttcaggttg tctaactcct tccttttcgg ttagagcgga tgtgggggga
     5581 gggcgtgaac gtaagcgtga cataactaat tacatgactc gagaagagat cacaccttcc
     5641 tcttcttctt ggggtcagcc ctgctgtctc caccgagctg agagaggtcg attcttgttt
     5701 catagagccc cgtaattgac tgatgaatca gtgtggcgtc caggacctcc tttgtagagg
     5761 tgtaccgctt tctgtctatg gtggtgtcga agtacttgaa ggctgcaggc gcgcccaagt
     5821 tggtcagagt aaacaagtag ataatgtttt ctgcctgctc cctgatgggc ttatccctgt
     5881 gcttattgta agcagaaagc accttatcga ggttagcgtc ggcgaggatc actcttttgg
     5941 agaattcgct tatttgctcg atgatctcat caaggtagtg tttgtgttgt tccacgaaca
     6001 gctgcttctg ctcattatct tcgggagacc ctttgagctt ttcatagtgg ctggccagat
     6061 acaagaaatt aacgtattta gagggcagtg ccagctcgtt acctttctgc agctcgcccg
     6121 cactagcgag cattcgtttc cggccgtttt caagctcaaa gagagagtac ttgggaagct
     6181 taatgatgag gtcttttttg acctctttat atcctttcgc ctcgagaaag tcgatggggt
     6241 ttttttcgaa gcttgatcgc tccatgattg tgatgcccag cagttccttg acgcttttga
     6301 gttttttaga cttccctttc tccactttgg ccacaaccag tacactgtaa gcgactgtag
     6361 gagaatcgaa tccgccgtat ttcttggggt cccaatcttt tttgcgtgcg atcagcttgt
     6421 cgctgttcct tttcgggagg atactttcct tggagaagcc tccggtctgt acttcggtct
     6481 ttttaacgat gttcacctgc ggcatggaca ggaccttccg gactgtcgcg aaatccctac
     6541 ccttgtccca cacgatttct cctgtttctc cgtttgtttc gataagtggt cgcttccgaa
     6601 tctctccatt ggccagtgta atctcggtct tgaaaaaatt cataatattg ctgtaaaaga
     6661 agtacttagc ggtggccttg cctatttcct gctcagactt tgcgatcatt ttcctaacat
     6721 cgtacacttt atagtctccg taaacaaatt cagattcaag cttgggatat tttttgataa
     6781 gtgcagtgcc taccactgca ttcaggtagg catcatgcgc atggtggtaa ttgttgatct
     6841 ctctcacctt ataaaactga aagtcctttc tgaaatctga gaccagctta gacttcagag
     6901 taataacttt cacctctcga atcagtttgt cattttcatc gtacttggtg ttcatgcgtg
     6961 aatcgagaat ttgggccacg tgcttggtga tctggcgtgt ctcaacaagc tgccttttga
     7021 tgaagccggc tttatccaac tcagacaggc cacctcgttc agccttagtc agattatcga
     7081 acttccgttg tgtgatcagt ttggcgttca gcagctgccg ccaataattt ttcattttct
     7141 tgacaacttc ttctgagggg acgttatcac tcttccctct atttttatcg gatcttgtca
     7201 acactttatt atcaatagaa tcatctttga gaaaagactg gggcacgata tgatccacgt
     7261 cgtagtcgga gagccgattg atgtccagtt cctgatccac gtacatgtcc ctgccgttct
     7321 gcaggtagta caggtagagc ttctcattct gaagctgggt gttttcaact gggtgttcct
     7381 taaggatttg ggaccccagt tcttttatac cctcttcaat cctcttcatc ctttccctac
     7441 tgttcttctg tcccttctgg gtagtttggt tctctcgggc catctcgata acgatattct
     7501 cgggcttatg ccttcccatt actttgacga gttcatccac gaccttaacg gtctgcagta
     7561 ttcccttttt gatagctggg ctacctgcaa gattagcgat gtgctcgtga agactgtccc
     7621 cctggccaga aacttgtgct ttctggatgt cctccttaaa ggtgagagag tcatcatgga
     7681 tcaactgcat gaagttccgg ttggcaaatc catcggactt aagaaaatcc aggattgtct
     7741 ttccactctg cttgtctcgg atcccattga tcagttttct tgacagccgc ccccatcctg
     7801 tatatcggcg cctcttgagc tgtttcatga ctttgtcgtc gaagagatga gcgtaagttt
     7861 tcaagcgttc ttcaatcatc tccctatctt caaacaacgt aagggtgagg acaatgtcct
     7921 caagaatgtc ctcgttctcc tcattgtcca ggaagtcctt gtctttaatg attttcagga
     7981 gatcgtgata cgttcccagg gatgcgttga agcgatcctc cactccgctg atttcaacag
     8041 agtcgaaaca ttcaatcttt ttgaaatagt cttctttgag ctgtttcacg gtaactttcc
     8101 ggttcgtctt gaagaggagg tccacgatag ctttcttctg ctctccagac aggaatgctg
     8161 gctttctcat cccttctgtg acgtatttga ccttggtgag ctcgttataa actgtgaagt
     8221 actcgtacag cagagagtgt ttaggaagca ccttttcgtt aggcagattt ttatcaaagt
     8281 tagtcatcct ttcgatgaag gactgggcag aggccccctt atccacgact tcctcgaagt
     8341 tccagggagt gatggtctct tctgatttgc gagtcatcca cgcgaatctg gaatttcccc
     8401 gggcgagggg gcctacatag tagggtatcc gaaatgtgag gattttctca atcttttccc
     8461 tgttatcttt caaaaagggg tagaaatcct cttgccgcct gaggatagcg tgcagttcgc
     8521 ccaggtgaat ctggtggggg atgcttccat tgtcgaaagt gcgctgtttg cgcaacagat
     8581 cttctctgtt aagctttacc agcagctcct cggtgccgtc cattttttcc aagatgggct
     8641 taataaattt gtaaaattcc tcctggcttg ctccgccgtc aatgtatccg gcgtagccat
     8701 ttttagactg atcgaagaaa atttccttgt acttctcagg cagttgctgt ctgacaaggg
     8761 ccttcagcaa agtcaagtct tggtggtgct catcatagcg cttgatcata ctagcgctca
     8821 gcggagcttt ggtgatctcc gtgttcactc gcagaatatc actcagcaga atggcgtctg
     8881 acaggttctt tgccgccaaa aaaaggtctg cgtactggtc gccgatctgg gccagcagat
     8941 tgtcgagatc atcatcgtag gtgtctttgc tcagttgaag cttggcatct tcggccaggt
     9001 cgaagttaga tttaaagttg ggggtcagcc cgagtgacag ggcgataaga ttaccaaaca
     9061 ggccgttctt cttctcccca gggagctgtg cgatgaggtt ttcgagccgc cgggatttgg
     9121 acagcctagc gctcaggatt gctttggcgt caactccgga tgcgttgatc gggttctctt
     9181 cgaaaagctg attgtaagtc tgaaccagtt ggataaagag tttgtcgaca tcgctgttgt
     9241 ctgggttcag gtccccctcg atgaggaagt gtccccgaaa tttgatcata tgcgccagcg
     9301 cgagatagat caaccgcaag tcagccttat cagtactgtc tacaagcttc ttcctcagat
     9361 gatatatggt tgggtacttt tcatggtacg ccacctcgtc cacgatattg ccaaagattg
     9421 ggtggcgctc gttcttttta tcctcctcca ccaaaaagga ctcctccagc ctatggaaga
     9481 aagagtcatc caccttagcc atctcattac taaagatctc ctgcaggtag cagatccgat
     9541 tctttctgcg ggtatatctg cgccgtgctg ttcttttgag ccgcgtggct tcggccgtct
     9601 ccccggagtc gaacaggagg gcgccaatga ggttcttctt tatgctgtgg cgatcggtat
     9661 tgcccagaac tttgaatttt ttgctcggca ccttgtactc gtccgtaatg acggcccaac
     9721 cgacgctgtt tgtgccgata tcgagcccaa tggagtactt cttgtccatt tttttccaag
     9781 ttgattgtat gcttggtata gcttgaaata ttgtgcagaa aaagaaacaa ggaagaaagg
     9841 gaacgagaac aatgacgagg aaacaaaaga ttaataattg caggtctatt tatacttgat
     9901 agcaagacag caaacttttt tttatttcaa attcaagtaa ctggaaggaa ggccgtatac
     9961 cgttgctcat tagagagtag tgtgcgtgaa tgaaggaagg aaaaagtttc gtgtgcttcg
    10021 agatacccct catcagctct ggaacaacga catctgttgg tgctgtcttt gtcgttaatt
    10081 ttttccttta gtgtcttcca tcattttttt gtcattgcgg atatggtgag acaacaacgg
    10141 gggagagaga aaagaaaaaa aaagaaaaga agttgcatgc gcctattatt acttcaatag
    10201 atggcaaatg gaaaaagggt agtgaaactt cgatatgatg atggctatca agtctagggc
    10261 tacagtatta gttcgttatg taccaccatc aatgaggcag tgtaattggt gtagtcttgt
    10321 ttagcccatt atgtcttgtc tggtatctgt tctattgtat atctcccctc cgccacctac
    10381 atgttaggga gaccaacgaa ggtattatag gaatcccgat gtatgggttt ggttgccaga
    10441 aaagaggaag tccatattgt acacccggaa acaacaaaag gatgtcgact atccactaga
    10501 cagaagtttg cgttccatac tgtcagaggt ggcattttac ataacaatag tgacaaaaaa
    10561 taaaaaaagc accgactcgg tgccactttt tcaagttgat aacggactag ccttatttta
    10621 acttgctatt tctagctcta aaacgcggcc ccagtatgaa aaccaaagtc ccattcgcca
    10681 cccgaaggtg ttgcccagcc ggcgccagcg aggaggctgg gaccatgccg gccatctctc
    10741 ccgggggcga gtcgaacgcc cgatctcaag atttcgtagt gataaattac agtcttgcgc
    10801 cttaaaccaa cttggctacc gagagtattt aattgttgaa gaaagagtat actagctcca
    10861 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
    10921 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
    10981 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
    11041 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
    11101 cgggga
//
