LOCUS       sequence-449556-       12840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Disruption of LEU2 genes in the Saccharomyces sense strict group 
            species.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12840)
  AUTHORS   Kiyokawa K, Sakuma T, Moriguchi K, Sugiyama M, Akao T, Yamamoto T, 
            Suzuki K
  TITLE     Conversion of polyploid and alloploid Saccharomyces sensu stricto 
            strains to leu2 mutants by genome DNA editing.
  JOURNAL   Appl Microbiol Biotechnol. 2024 Jul 12;108(1):416. doi: 
            10.1007/s00253-024-13242-y.
  PUBMED    38995331
REFERENCE   2  (bases 1 to 12840)
  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"; journalName: "Appl 
            Microbiol Biotechnol."; date: "2024-07-12"; volume: "108(1):416. 
            doi"; pages: " 10.1007/s00253-024-13242-y"
COMMENT     Sequence Label: pYAMTr2GCsgSc-SeLEU2
FEATURES             Location/Qualifiers
     source          1..12840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      29..1375
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     1416..1434
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1472..1494)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1594..1613
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     gene            1669..3025
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1669..2012
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2013..2822
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2080..2099)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2828..3025
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(3107..3562)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3244..3265)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3456..3475
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3688..3710
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3702..3719
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3703..3719
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3726..3745
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3726..3744
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3777..3793
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3777..3793
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    3867..3888
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3903..3908,3909..3926,3927..3933)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3941..3957
                     /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     3965..3981
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4002..4020
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4051..4319
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        4340..4414
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      4419..4438
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     terminator      complement(4556..4743)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     4664..4688
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(4771..4791)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(7186..8904)
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     regulatory      8901..8910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(8939..9605)
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     complement(9046..9069)
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(9650..9666)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9651..9667)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(9695..9713)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9734..9750)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9758..9774
                     /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(9782..9788,9789..9806,9807..9812))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_feature    9850..9874
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(10141..10935)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(10252..10271)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     10862..10881
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      11349..12120
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             12121..12783
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
        1 gtccttttca tcacgtgagt gccacctgaa cgaagcatct gtgcttcatt ttgtagaaca
       61 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
      121 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta
      181 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
      241 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
      301 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact
      361 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
      421 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
      481 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
      541 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
      601 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
      661 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
      721 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
      781 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
      841 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
      901 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtct
      961 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat
     1021 aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa
     1081 cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt
     1141 atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg cgtacttata
     1201 tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt atcccattcc
     1261 atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata tgctgccact
     1321 cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt ggatcatact
     1381 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     1441 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     1501 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     1561 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     1621 tgcaccatag tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc
     1681 cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc
     1741 ccgtacattt agcccataca tccccatgta taatcatttg catccataca ttttgatggc
     1801 cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca gggaaacgct
     1861 cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt
     1921 taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac
     1981 agttctcaca tcacatccga acataaacaa ccatgggtaa ggaaaagact cacgtttcga
     2041 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     2101 atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt
     2161 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     2221 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     2281 atgatgcatg gttactcacc actgcgatcc ccggcaaaac agcattccag gtattagaag
     2341 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     2401 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     2461 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     2521 gctggcctgt tgaacaagtc tggaaagaaa tgcataagct tttgccattc tcaccggatt
     2581 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     2641 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     2701 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     2761 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     2821 aatcagtact gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt
     2881 ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct
     2941 cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg
     3001 tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccagtgtc
     3061 gaaaacaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt
     3121 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3181 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3241 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3301 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3361 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3421 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3481 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3541 aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc
     3601 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     3661 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     3721 aattgtaata cgactcacta tagggcgaat tggagctcca ccgcggtggc ggccgctcta
     3781 gaactagtgg atcccccggg ctgcaggaat tcgatctggc acgacaggtt tcccgactgg
     3841 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     3901 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     3961 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     4021 gaacaaaagc tggagctcca ccgcggacaa tctttgaaaa gataatgtat gattatgctt
     4081 tcactcatat ttatacagaa acttgatgtt ttctttcgag tatatacaag gtgattacat
     4141 gtacgtttga agtacaactc tagattttgt agtgccctct tgggctagcg gtaaaggtgc
     4201 gcattttttc acaccctaca atgttctgtt caaaagattt tggtcaaacg ctgtagaagt
     4261 gaaagttggt gcgcatgttt cggcgttcga aacttctccg cagtgaaaga taaatgatct
     4321 gctgtgggtg gtcctaaatg ttttagagct agaaatagca agttaaaata aggctagtcc
     4381 gttatcaact tgaaaaagtg gcaccgagtc ggtggtgctt tttttgtttt ttatgtcttc
     4441 gagtcatgta attagttata agcatgtgag ctaagacact gtaattgcca atctaaacga
     4501 taccacggcc gctctagaga aatggggagc gatttgcagg catttgctcg gcatgccggt
     4561 agaggtgtgg tcaataagag cgacctcatg ctatacctga gaaagcaacc tgacctacag
     4621 gaaagagtta ctcaagaata agaattttcg ttttaaaacc taagagtcac tttaaaattt
     4681 gtatacactt atttttttta taacttattt aataataaaa atcataaatc ataagaaatt
     4741 cgcctcgaga gtctaggatc cggaactacc taccttgcgc tttttcttgg gagctccctc
     4801 atcccctccg agctgtgaga ggtcaatccg agtttcgtaa agtccggtga tgctctggtg
     4861 gatgagggtg gcgtccagca cttctttggt ggaggtatag cgtttccggt cgatggtggt
     4921 gtcgaagtac ttgaatgcgg caggggcacc aagattggtg agggtgaaca ggtggatgat
     4981 attctcagcc tgctcccgaa ttggtttgtc tctgtgctta ttgtatgcgc taaggacttt
     5041 atcgaggttg gcatcagcga gaatgacgcg tttagaaaac tcggaaatct gctcaatgat
     5101 ctcgtccaga taatgcttat gctgctccac aaacagttgc ttttgctcgt tgtcctcggg
     5161 gctccccttc agcttctcgt aatggcttgc aaggtagagg aaattcacgt acttggaggg
     5221 gagagccagc tcattaccct tctgaagttc gccagcggag gcgagcatcc gcttccgccc
     5281 attctccagt tcgaacaggc tatactttgg gagcttgatg ataaggtcct tcttgacttc
     5341 cttgtaacct ttggcttcga gaaagtcaat tgggttcttc tcgaaggagc tgcgctccat
     5401 aatggtgata cccagcagtt ccttcacgct cttgagcttc ttagactttc ctttttccac
     5461 cttagccacg accaggacag aataagccac agttggtgaa tcgaatccgc cgtacttctt
     5521 agggtcccaa tccttcttcc ttgcaatgag cttgtcggag ttgcgcttgg ggaggattga
     5581 ttccttagaa aagcctccgg tttgcacctc ggttttcttc acgatgttca cttgaggcat
     5641 agaaagcact ttgcgcacgg tagcgaaatc tctgcccttg tcccacacga tttcgccagt
     5701 ctccccatta gtctcgatga gcggcctctt ccggatctca ccgtttgcaa gggtgatttc
     5761 ggtcttgaag aaattcatga tgttgctgta gaagaagtac tttgcggtgg cctttccgat
     5821 ctcctgctca gatttggcaa tcatcttgcg cacgtcgtag accttgtagt ctccgtacac
     5881 gaactcgctt tccagctttg ggtacttctt gattagggcg gttcccacca cggcattcag
     5941 atatgcgtca tgagcatgat gatagttatt gatctcccgc actttgtaga attgaaagtc
     6001 ctttctgaaa tcgctgacca gcttggactt cagggtaatc accttcacct cgcggataag
     6061 tttgtcgttt tcgtcgtact tggtattcat gcgtgagtca aggatttggg ctacatgttt
     6121 agtaatctgc cgagtctcca caagttgccg tttaatgaat cctgctttgt ccagttcgct
     6181 cagtccgccc ctctcagcct tagtgagatt gtcgaacttc cgctgagtga ttagctttgc
     6241 gttcagaagc tggcgccaat agttcttcat ctttttcacg acttcctctg agggcacgtt
     6301 atctgacttt ccccggttct tgtctgagcg agtcagcacc ttattgtcaa ttgagtcgtc
     6361 tttgaggaag ctctgaggga cgatgtggtc cacgtcgtag tctgagagcc tgttgatatc
     6421 aagctcttgg tccacataca tatcgcgtcc attttgaagg tagtacagat agagcttctc
     6481 attctgaagc tgggtgtttt ccaccgggtg ctctttaagg atctgtgacc cgagctcctt
     6541 aatcccttcc tcaattcttt tcatccgctc cctgctgttc ttctgtccct tctgggtagt
     6601 ctggttttcc ctggccatct cgatgacaat gttctctggt ttatggcgac ccatcacctt
     6661 gaccagctcg tccacgactt tcacggtctg gagaataccc ttcttaatag cgggtgaacc
     6721 agccagattt gcgatatgct cgtgcagtga gtcaccttgt ccagacactt gggccttctg
     6781 gatgtcctcc ttaaaggtca gggaatcatc atggataagt tgcatgaagt tgcgattagc
     6841 gaatccgtcg ctcttaagga aatcaagaat ggtctttccg ctctgtttgt cccggattcc
     6901 gttgatgagt ttgcggctaa ggcgtcccca tccggtatat cttctccgtt taagttgctt
     6961 catcacttta tcgtcgaaca gatgggcgta ggtcttaagc ctctcctcga tcatttcccg
     7021 gtcctcgaac agagtgagag tcaggacaat gtcctcaagg atgtcctctt tctcctcatt
     7081 gtccagaaaa tccttgtcct tgatgatctt aagcagatca tggtaggtac ccaggcttgc
     7141 gttgaaccga tcttcgaccc cgctaatctc cacagagtcg aagcattcaa ttttcttgaa
     7201 atagtcctcc ttcagctgct tcacggtcac ctttctattg gtcttgaaca gaaggtccac
     7261 aattgccttc ttttgttcgc ctgacaggaa tgcgggcttc ctcatccctt cggtgacata
     7321 tttcaccttg gtcagctcgt tgtacacggt aaagtactcg tagagcaggg aatgtttcgg
     7381 caggaccttc tcattgggga ggttcttatc gaagttagtc atccgttcga tgaatgactg
     7441 agcgctggct cccttgtcca ccacctcttc gaagttccag ggagtgatag tttcttctga
     7501 ctttctggtc atccaagcaa accggctatt tcctctggcg aggggtccca cgtagtaggg
     7561 gatgcggaaa gtgagaattt tctcaatctt ctccctgttg tccttaagaa aggggtagaa
     7621 gtcctcttgg cgccgaagga tggcgtgaag ctccccaaga tggatctggt ggggaatgct
     7681 cccattgtcg aaagtgcgct gcttccgcag cagatcctcc cgattcagtt tcacgaggag
     7741 ctcttcggtg ccgtccatct tctcaaggat aggtttgata aacttataaa attcctcttg
     7801 ggaggcacct ccgtcgatgt agccggcgta tccattcttt gactggtcaa agaagatttc
     7861 cttgtacttc tccggaagct gctgccggac gagtgcctta agcagggtca ggtcctggtg
     7921 gtgctcatca taccgcttaa tcattgaagc tgaaagaggg gccttggtga tttcagtgtt
     7981 cacgcggaga atgtcagaca gcaggatggc gtcagaaaga ttcttagcag caaggaaaag
     8041 gtcggcgtat tggtcaccga tctgggccag gaggttgtcg agatcgtcat cataggtgtc
     8101 cttggacagt tgcagcttgg catcctctgc caggtcgaaa ttagacttga aattgggagt
     8161 cagcccaagt gaaagagcaa tcaggttccc gaacagtcca tttttctttt cacccggcag
     8221 ctgagcgata agattctcaa gtctgcggct ctttgacagc cgggctgaca gaatggcctt
     8281 ggcatcgact ccgcttgcat tgatggggtt ctcttcaaag agctggttgt aagtctgcac
     8341 cagctggatg aacagtttat ccacatcgct attgtcagga ttgaggtcgc cctcgatcag
     8401 gaagtggcct ctgaacttaa tcatgtgggc cagggccagg tagataagcc ggaggtctgc
     8461 cttgtcggtt gagtccacca gctttttgcg aagatggtag atggttgggt atttctcgtg
     8521 ataggccacc tcgtcgacga tattaccgaa gatgggatgc cgttcatgct tcttatcctc
     8581 ttccacgagg aatgattctt ccagcctatg gaagaagctg tcgtcgacct ttgccatttc
     8641 gttggagaat atctcttgaa ggtaacaaat ccggttcttc ctcctggtgt accgccgtct
     8701 agcagtccgc ttgaggcggg ttgcttcagc ggtctcccct gaatcaaaga ggagagctcc
     8761 gatcagattt ttcttgatgg aatgccggtc ggtgttcccg agcaccttga acttcttaga
     8821 gggcaccttg tactcgtcag tgatgacggc ccacccgacg ctattagtcc cgatgtccag
     8881 tccgatagaa tacttcttgt ccatggtggc gagatctaat tcgatccata tcagatcttt
     8941 tgtttgttta tgtgtgttta ttcgaaacta agttcttggt gttttaaaac taaaaaaaag
     9001 actaactata aaagtagaat ttaagaagtt taagaaatag atttacagaa ttacaatcaa
     9061 tacctaccgt ctttatatac ttattagtca agtaggggaa taatttcagg gaactggttt
     9121 caaccttttt tttcagcttt ttccaaatca gagagagcag aaggtaatag aaggtgtaag
     9181 aaaatgagat agatacatgc gtgggtcaat tgccttgtgt catcatttac tccaggcagg
     9241 ttgcatcact ccattgaggt tgtgcccgtt ttttgcctgt ttgtgcccct gttctctgta
     9301 gttgcgctaa gagaatggac ctatgaactg atggttggtg aagaaaacaa tattttggtg
     9361 ctgggattct ttttttttct ggatgccagc ttaaaaagcg ggctccatta tatttagtgg
     9421 atgccaggaa taaactgttc acccagacac ctacgatgtt atatattctg tgtaacccgc
     9481 cccctatttt gggcatgtac gggttacagc agaattaaaa ggctaatttt ttgactaaat
     9541 aaagttagga aaatcactac tattaattat ttacgtattc tttgaaatgg cagtattgat
     9601 aatgataaac tcgaactgac tagtagactg aattcgatat caagcttatc gataccgtcg
     9661 acctcgaggg ggggcccggt accagctttt gttcccttta gtgagggtta atttcgagct
     9721 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     9781 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgaggtacg
     9841 gatcgatcct gacaggatat attggcgggt aaacctaaga gaaaagagcg tttattagaa
     9901 taatcggata tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg tatgtgcatg
     9961 ccaaccacag ggttccccag atctggcgcc ggccagcgag acgagcaaga ttggccgcgg
    10021 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
    10081 aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaacccc agagtcccgc
    10141 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
    10201 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
    10261 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
    10321 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
    10381 gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc
    10441 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
    10501 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
    10561 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
    10621 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
    10681 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
    10741 cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
    10801 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
    10861 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
    10921 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct
    10981 gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac
    11041 cttaccagag ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca
    11101 gtctagctat cgccatgtaa gcccactgcc tcgagactag ttggtgtcca accggctcga
    11161 cgggggcagc gcaaggcggt gcctccggcg ggccactcaa tgcttgagta tactcactag
    11221 actttgcttc gcaaagtcgt gaccgcctac ggcggctgcg gcgccctacg ggcttgctct
    11281 ccgggcttcg ccctgcgcgg tcgctgcgct cccttgccag cccgtggata tgtggacgat
    11341 ggccgcgagc ggccaccggc tggctcgctt cgctcggccc gtggacaacc ctgctggaca
    11401 agctgatgga caggctgcgc ctgcccacga gcttgaccac agggattgcc caccggctac
    11461 ccagccttcg accacatacc caccggctcc aactgcgcgg cctgcggcct tgccccatca
    11521 atttttttaa ttttctctgg ggaaaagcct ccggcctgcg gcctgcgcgc ttcgcttgcc
    11581 ggttggacac caagtggaag gcgggtcaag gctcgcgcag cgaccgcgca gcggcttggc
    11641 cttgacgcgc ctggaacgac ccaagcctat gcgagtgggg gcagtcgaag ggcgaagccc
    11701 gcccgcctgc cccccgagcc tcacggcggc gagtgcgggg gttccaaggg ggcagcgcca
    11761 ccttgggcaa ggccgaaggc cgcgcagtcg atcaacaagc cccggagggg ccactttttg
    11821 ccggaggggg agccgcgccg aaggcgtggg ggaaccccgc aggggtgccc ttctttgggc
    11881 accaaagaac tagatatagg gcgaaatgcg aaagacttaa aaatcaacaa cttaaaaaag
    11941 gggggtacgc aacagctcat tgcggcaccc cccgcaatag ctcattgcgt aggttaaaga
    12001 aaatctgtaa ttgactgcca cttttacgca acgcataatt gttgtcgcgc tgccgaaaag
    12061 ttgcagctga ttgcgcatgg tgccgcaacc gtgcggcacc cctaccgcat ggagataagc
    12121 atggccacgc agtccagaga aatcggcatt caagccaaga acaagcccgg tcactgggtg
    12181 caaacggaac gcaaagcgca tgaggcgtgg gccgggctta ttgcgaggaa acccacggcg
    12241 gcaatgctgc tgcatcacct cgtggcgcag atgggccacc agaacgccgt ggtggtcagc
    12301 cagaagacac tttccaagct catcggacgt tctttgcgga cggtccaata cgcagtcaag
    12361 gacttggtgg ccgagcgctg gatctccgtc gtgaagctca acggccccgg caccgtgtcg
    12421 gcctacgtgg tcaatgaccg cgtggcgtgg ggccagcccc gcgaccagtt gcgcctgtcg
    12481 gtgttcagtg ccgccgtggt ggttgatcac gacgaccagg acgaatcgct gttggggcat
    12541 ggcgacctgc gccgcatccc gaccctgtat ccgggcgagc agcaactacc gaccggcccc
    12601 ggcgaggagc cgcccagcca gcccggcatt ccgggcatgg aaccagacct gccagccttg
    12661 accgaaacgg aggaatggga acggcgcggg cagcagcgcc tgccgatgcc cgatgagccg
    12721 tgttttctgg acgatggcga gccgttggag ccgccgacac gggtcacgct gccgcgccgg
    12781 tagcacttgg gttgcgcagc aacccgtaag tgcgctgttc cagactatcg gctgtagccg
//
