LOCUS       sequence-116839-       11928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High copy Cas9 expression vector with LEU2 marker for yeast 
            transformation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11928)
  AUTHORS   Laughery MF, Hunter T, Brown A, Hoopes J, Ostbye T, Shumaker T, 
            Wyrick JJ
  TITLE     New vectors for simple and streamlined CRISPR-Cas9 genome editing in
            Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2015 Dec;32(12):711-20. doi: 10.1002/yea.3098. Epub 2015 Sep 
            21.
  PUBMED    26305040
REFERENCE   2  (bases 1 to 11928)
  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: "10.1002/yea.3098"; 
            journalName: "Yeast"; date: "2015-12"; volume: "32"; issue: "12"; 
            pages: "711-20"
COMMENT     Sequence Label: pJH001
FEATURES             Location/Qualifiers
     source          1..11928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(664..1758)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1759..2166)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2467..2922)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2604..2625)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2816..2835
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3048..3070
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3062..3079
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3063..3079
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3089..3108
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3089..3107
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3132..3148
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3133..3149
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3194..3860
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3730..3753
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     regulatory      3889..3898
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3895..7998
                     /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
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             3895..7998
                     /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
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8008..8028
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8056..8243
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(8111..8135)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(8323..8343)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8323..8341)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8362..8378)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8362..8378)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8375..8397)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8386..8402
                     /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(8410..8416,8417..8434,8435..8440))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8455..8476
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8593..8610)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8764..9352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8844..8863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9523..10314,10315..10383))
                     /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     10146..10165
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10384..10488)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      10515..11861
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     11901..11919
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatcga ctacgtcgtt aaggccgttt ctgacagagt aaaattcttg agggaacttt
      241 caccattatg ggaaatggtt caagaaggta ttgacttaaa ctccatcaaa tggtcaggtc
      301 attgagtgtt ttttatttgt tgtatttttt tttttttaga gaaaatcctc caatatataa
      361 attaggaatc atagtttcat gattttctgt tacacctaac tttttgtgtg gtgccctcct
      421 ccttgtcaat attaatgtta aagtgcaatt ctttttcctt atcacgttga gccattagta
      481 tcaatttgct tacctgtatt cctttacatc ctcctttttc tccttcttga taaatgtatg
      541 tagattgcgt atatagtttc gtctacccta tgaacatatt ccattttgta atttcgtgtc
      601 gtttctatta tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc
      661 tttttaagca aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact
      721 gttggaacca cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc
      781 ttcaatggcc ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga
      841 caagatagtg gcgatagggt tgaccttatt ctttggcaaa tctggagcag aaccgtggca
      901 tggttcgtac aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg
      961 caacaaaccc aaggaacctg ggataacgga ggcttcatcg gagatgatat caccaaacat
     1021 gttgctggtg attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc
     1081 agaatcaatc aattgatgtt gaaccttcaa tgtaggaaat tcgttcttga tggtttcctc
     1141 cacagttttt ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat
     1201 aggcaatggt ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac
     1261 ttctggaacg gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct
     1321 cttaccaaag taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc
     1381 aaattgtggc ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct
     1441 taagttggcg tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac
     1501 actacctgta ccccatttag gaccacccac agcacctaac aaaacggcat cagccttctt
     1561 ggaggcttcc agcgcctcat ctggaagtgg aacacctgta gcatctatag cagcaccacc
     1621 aattaaatga ttttcgaaat cgaacttgac attggaacga acatcagaaa tagctttaag
     1681 aaccttaatg gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat
     1741 cttcttaggg gcagacatta gaatggtata tccttgaaat atatatatat atattgctga
     1801 aatgtaaaag gtaagaaaag ttagaaagta agacgattgc taaccaccta ttggaaaaaa
     1861 caataggtcc ttaaataata ttgtcaactt caagtattgt gatgcaagca tttagtcatg
     1921 aacgcttctc tattctatat gaaaagccgg ttccggcgct ctcacctttc ctttttctcc
     1981 caatttttca gttgaaaaag gtatatgcgt caggcgacct ctgaaattaa caaaaaattt
     2041 ccagtcatcg aatttgattc tgtgcgatag cgcccctgtg tgttctcgtt atgttgagga
     2101 aaaaaataat ggttgctaag agattcgaac tcttgcatct tacgatacct gagtattccc
     2161 acagttaact gcggtcaaga tatttcttga atcaggcgcc ttagaccgct cggccaaaca
     2221 accaattact tgttgagaaa tagagtataa ttatcctata aatataacgt ttttgaacac
     2281 acatgaacaa ggaagtacag gacaattgat tttgaagaga atgtggattt tgatgtaatt
     2341 gttgggattc catttttaat aaggcaataa tattaggtat gtagatatac tagaagttct
     2401 cctcgaccgg tcgatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     2461 atcaggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2521 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2581 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2641 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2701 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2761 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2821 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2881 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct
     2941 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     3001 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     3061 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac
     3121 cgggcccccc ctcgaggtcg acggtatcga taagcttgat atcgaattca gtctactagt
     3181 cagttcgagt ttatcattat caatactgcc atttcaaaga atacgtaaat aattaatagt
     3241 agtgattttc ctaactttat ttagtcaaaa aattagcctt ttaattctgc tgtaacccgt
     3301 acatgcccaa aatagggggc gggttacaca gaatatataa catcgtaggt gtctgggtga
     3361 acagtttatt cctggcatcc actaaatata atggagcccg ctttttaagc tggcatccag
     3421 aaaaaaaaag aatcccagca ccaaaatatt gttttcttca ccaaccatca gttcataggt
     3481 ccattctctt agcgcaacta cagagaacag gggcacaaac aggcaaaaaa cgggcacaac
     3541 ctcaatggag tgatgcaacc tgcctggagt aaatgatgac acaaggcaat tgacccacgc
     3601 atgtatctat ctcattttct tacaccttct attaccttct gctctctctg atttggaaaa
     3661 agctgaaaaa aaaggttgaa accagttccc tgaaattatt cccctacttg actaataagt
     3721 atataaagac ggtaggtatt gattgtaatt ctgtaaatct atttcttaaa cttcttaaat
     3781 tctactttta tagttagtct tttttttagt tttaaaacac caagaactta gtttcgaata
     3841 aacacacata aacaaacaaa agatctgata tggatcgaat tagatctcgc caccatggac
     3901 aagaagtatt ctatcggact ggacatcggg actaatagcg tcgggtgggc cgtgatcact
     3961 gacgagtaca aggtgccctc taagaagttc aaggtgctcg ggaacaccga ccggcattcc
     4021 atcaagaaaa atctgatcgg agctctcctc tttgattcag gggagaccgc tgaagcaacc
     4081 cgcctcaagc ggactgctag acggcggtac accaggagga agaaccggat ttgttacctt
     4141 caagagatat tctccaacga aatggcaaag gtcgacgaca gcttcttcca taggctggaa
     4201 gaatcattcc tcgtggaaga ggataagaag catgaacggc atcccatctt cggtaatatc
     4261 gtcgacgagg tggcctatca cgagaaatac ccaaccatct accatcttcg caaaaagctg
     4321 gtggactcaa ccgacaaggc agacctccgg cttatctacc tggccctggc ccacatgatc
     4381 aagttcagag gccacttcct gatcgagggc gacctcaatc ctgacaatag cgatgtggat
     4441 aaactgttca tccagctggt gcagacttac aaccagctct ttgaagagaa ccccatcaat
     4501 gcaagcggag tcgatgccaa ggccattctg tcagcccggc tgtcaaagag ccgcagactt
     4561 gagaatctta tcgctcagct gccgggtgaa aagaaaaatg gactgttcgg gaacctgatt
     4621 gctctttcac ttgggctgac tcccaatttc aagtctaatt tcgacctggc agaggatgcc
     4681 aagctgcaac tgtccaagga cacctatgat gacgatctcg acaacctcct ggcccagatc
     4741 ggtgaccaat acgccgacct tttccttgct gctaagaatc tttctgacgc catcctgctg
     4801 tctgacattc tccgcgtgaa cactgaaatc accaaggccc ctctttcagc ttcaatgatt
     4861 aagcggtatg atgagcacca ccaggacctg accctgctta aggcactcgt ccggcagcag
     4921 cttccggaga agtacaagga aatcttcttt gaccagtcaa agaatggata cgccggctac
     4981 atcgacggag gtgcctccca agaggaattt tataagttta tcaaacctat ccttgagaag
     5041 atggacggca ccgaagagct cctcgtgaaa ctgaatcggg aggatctgct gcggaagcag
     5101 cgcactttcg acaatgggag cattccccac cagatccatc ttggggagct tcacgccatc
     5161 cttcggcgcc aagaggactt ctaccccttt cttaaggaca acagggagaa gattgagaaa
     5221 attctcactt tccgcatccc ctactacgtg ggacccctcg ccagaggaaa tagccggttt
     5281 gcttggatga ccagaaagtc agaagaaact atcactccct ggaacttcga agaggtggtg
     5341 gacaagggag ccagcgctca gtcattcatc gaacggatga ctaacttcga taagaacctc
     5401 cccaatgaga aggtcctgcc gaaacattcc ctgctctacg agtactttac cgtgtacaac
     5461 gagctgacca aggtgaaata tgtcaccgaa gggatgagga agcccgcatt cctgtcaggc
     5521 gaacaaaaga aggcaattgt ggaccttctg ttcaagacca atagaaaggt gaccgtgaag
     5581 cagctgaagg aggactattt caagaaaatt gaatgcttcg actctgtgga gattagcggg
     5641 gtcgaagatc ggttcaacgc aagcctgggt acctaccatg atctgcttaa gatcatcaag
     5701 gacaaggatt ttctggacaa tgaggagaac gaggacatcc ttgaggacat tgtcctgact
     5761 ctcactctgt tcgaggaccg ggaaatgatc gaggagaggc ttaagaccta cgcccatctg
     5821 ttcgacgata aagtgatgaa gcaacttaaa cggagaagat ataccggatg gggacgcctt
     5881 agccgcaaac tcatcaacgg aatccgggac aaacagagcg gaaagaccat tcttgatttc
     5941 cttaagagcg acggattcgc taatcgcaac ttcatgcaac ttatccatga tgattccctg
     6001 acctttaagg aggacatcca gaaggcccaa gtgtctggac aaggtgactc actgcacgag
     6061 catatcgcaa atctggctgg ttcacccgct attaagaagg gtattctcca gaccgtgaaa
     6121 gtcgtggacg agctggtcaa ggtgatgggt cgccataaac cagagaacat tgtcatcgag
     6181 atggccaggg aaaaccagac tacccagaag ggacagaaga acagcaggga gcggatgaaa
     6241 agaattgagg aagggattaa ggagctcggg tcacagatcc ttaaagagca cccggtggaa
     6301 aacacccagc ttcagaatga gaagctctat ctgtactacc ttcaaaatgg acgcgatatg
     6361 tatgtggacc aagagcttga tatcaacagg ctctcagact acgacgtgga ccacatcgtc
     6421 cctcagagct tcctcaaaga cgactcaatt gacaataagg tgctgactcg ctcagacaag
     6481 aaccggggaa agtcagataa cgtgccctca gaggaagtcg tgaaaaagat gaagaactat
     6541 tggcgccagc ttctgaacgc aaagctgatc actcagcgga agttcgacaa tctcactaag
     6601 gctgagaggg gcggactgag cgaactggac aaagcaggat tcattaaacg gcaacttgtg
     6661 gagactcggc agattactaa acatgtcgcc caaatccttg actcacgcat gaataccaag
     6721 tacgacgaaa acgacaaact tatccgcgag gtgaaggtga ttaccctgaa gtccaagctg
     6781 gtcagcgatt tcagaaagga ctttcaattc tacaaagtgc gggagatcaa taactatcat
     6841 catgctcatg acgcatatct gaatgccgtg gtgggaaccg ccctgatcaa gaagtaccca
     6901 aagctggaaa gcgagttcgt gtacggagac tacaaggtct acgacgtgcg caagatgatt
     6961 gccaaatctg agcaggagat cggaaaggcc accgcaaagt acttcttcta cagcaacatc
     7021 atgaatttct tcaagaccga aatcaccctt gcaaacggtg agatccggaa gaggccgctc
     7081 atcgagacta atggggagac tggcgaaatc gtgtgggaca agggcagaga tttcgctacc
     7141 gtgcgcaaag tgctttctat gcctcaagtg aacatcgtga agaaaaccga ggtgcaaacc
     7201 ggaggctttt ctaaggaatc aatcctcccc aagcgcaact ccgacaagct cattgcaagg
     7261 aagaaggatt gggaccctaa gaagtacggc ggattcgatt caccaactgt ggcttattct
     7321 gtcctggtcg tggctaaggt ggaaaaagga aagtctaaga agctcaagag cgtgaaggaa
     7381 ctgctgggta tcaccattat ggagcgcagc tccttcgaga agaacccaat tgactttctc
     7441 gaagccaaag gttacaagga agtcaagaag gaccttatca tcaagctccc aaagtatagc
     7501 ctgttcgaac tggagaatgg gcggaagcgg atgctcgcct ccgctggcga acttcagaag
     7561 ggtaatgagc tggctctccc ctccaagtac gtgaatttcc tctaccttgc aagccattac
     7621 gagaagctga aggggagccc cgaggacaac gagcaaaagc aactgtttgt ggagcagcat
     7681 aagcattatc tggacgagat cattgagcag atttccgagt tttctaaacg cgtcattctc
     7741 gctgatgcca acctcgataa agtccttagc gcatacaata agcacagaga caaaccaatt
     7801 cgggagcagg ctgagaatat catccacctg ttcaccctca ccaatcttgg tgcccctgcc
     7861 gcattcaagt acttcgacac caccatcgac cggaaacgct atacctccac caaagaagtg
     7921 ctggacgcca ccctcatcca ccagagcatc accggacttt acgaaactcg gattgacctc
     7981 tcacagctcg gaggggatga gggagctccc aagaaaaagc gcaaggtagg tagttccgga
     8041 tcctagactc tcgaggcgaa tttcttatga tttatgattt ttattattaa ataagttata
     8101 aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta
     8161 ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagca tgaggtcgct
     8221 cttattgacc acacctctac cggcatgccg agcaaatgcc tgcaaatcgc tccccatttc
     8281 tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat
     8341 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     8401 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     8461 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     8521 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     8581 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     8641 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     8701 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     8761 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     8821 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     8881 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     8941 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     9001 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     9061 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     9121 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     9181 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     9241 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     9301 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     9361 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     9421 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     9481 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     9541 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     9601 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     9661 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     9721 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     9781 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     9841 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     9901 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     9961 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
    10021 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
    10081 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
    10141 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
    10201 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
    10261 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
    10321 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    10381 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
    10441 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    10501 aaaagtgcca cctgaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga
    10561 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
    10621 cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa
    10681 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
    10741 caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa
    10801 tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg
    10861 tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa
    10921 gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg
    10981 tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta
    11041 tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat
    11101 tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat
    11161 aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa
    11221 tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag
    11281 atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat
    11341 atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt
    11401 agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg
    11461 gttttcaaaa gcgctctgaa gttcctatac tttctagcta gagaatagga acttcggaat
    11521 aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa atgcaacgcg agctgcgcac
    11581 atacagctca ctgttcacgt cgcacctata tctgcgtgtt gcctgtatat atatatacat
    11641 gagaagaacg gcatagtgcg tgtttatgct taaatgcgta cttatatgcg tctatttatg
    11701 taggatgaaa ggtagtctag tacctcctgt gatattatcc cattccatgc ggggtatcgt
    11761 atgcttcctt cagcactacc ctttagctgt tctatatgct gccactcctc aattggatta
    11821 gtctcatcct tcaatgctat catttccttt gatattggat catctaagaa accattatta
    11881 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
