LOCUS       sequence-449569-       12104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast genome editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12104)
  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 12104)
  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: pYAMTr2GCas
FEATURES             Location/Qualifiers
     source          1..12104
                     /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"
     terminator      complement(3820..4007)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     3928..3952
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(4035..4055)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(6450..8168)
                     /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      8165..8174
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(8203..8869)
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     complement(8310..8333)
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(8914..8930)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8915..8931)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(8959..8977)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8998..9014)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8998..9014)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9011..9033)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9022..9038
                     /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(9046..9052,9053..9070,9071..9076))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_feature    9114..9138
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(9405..10199)
                     /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(9516..9535)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     10126..10145
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      10613..11384
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             11385..12047
                     /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 gagaaatggg gagcgatttg caggcatttg ctcggcatgc cggtagaggt gtggtcaata
     3841 agagcgacct catgctatac ctgagaaagc aacctgacct acaggaaaga gttactcaag
     3901 aataagaatt ttcgttttaa aacctaagag tcactttaaa atttgtatac acttattttt
     3961 tttataactt atttaataat aaaaatcata aatcataaga aattcgcctc gagagtctag
     4021 gatccggaac tacctacctt gcgctttttc ttgggagctc cctcatcccc tccgagctgt
     4081 gagaggtcaa tccgagtttc gtaaagtccg gtgatgctct ggtggatgag ggtggcgtcc
     4141 agcacttctt tggtggaggt atagcgtttc cggtcgatgg tggtgtcgaa gtacttgaat
     4201 gcggcagggg caccaagatt ggtgagggtg aacaggtgga tgatattctc agcctgctcc
     4261 cgaattggtt tgtctctgtg cttattgtat gcgctaagga ctttatcgag gttggcatca
     4321 gcgagaatga cgcgtttaga aaactcggaa atctgctcaa tgatctcgtc cagataatgc
     4381 ttatgctgct ccacaaacag ttgcttttgc tcgttgtcct cggggctccc cttcagcttc
     4441 tcgtaatggc ttgcaaggta gaggaaattc acgtacttgg aggggagagc cagctcatta
     4501 cccttctgaa gttcgccagc ggaggcgagc atccgcttcc gcccattctc cagttcgaac
     4561 aggctatact ttgggagctt gatgataagg tccttcttga cttccttgta acctttggct
     4621 tcgagaaagt caattgggtt cttctcgaag gagctgcgct ccataatggt gatacccagc
     4681 agttccttca cgctcttgag cttcttagac tttccttttt ccaccttagc cacgaccagg
     4741 acagaataag ccacagttgg tgaatcgaat ccgccgtact tcttagggtc ccaatccttc
     4801 ttccttgcaa tgagcttgtc ggagttgcgc ttggggagga ttgattcctt agaaaagcct
     4861 ccggtttgca cctcggtttt cttcacgatg ttcacttgag gcatagaaag cactttgcgc
     4921 acggtagcga aatctctgcc cttgtcccac acgatttcgc cagtctcccc attagtctcg
     4981 atgagcggcc tcttccggat ctcaccgttt gcaagggtga tttcggtctt gaagaaattc
     5041 atgatgttgc tgtagaagaa gtactttgcg gtggcctttc cgatctcctg ctcagatttg
     5101 gcaatcatct tgcgcacgtc gtagaccttg tagtctccgt acacgaactc gctttccagc
     5161 tttgggtact tcttgattag ggcggttccc accacggcat tcagatatgc gtcatgagca
     5221 tgatgatagt tattgatctc ccgcactttg tagaattgaa agtcctttct gaaatcgctg
     5281 accagcttgg acttcagggt aatcaccttc acctcgcgga taagtttgtc gttttcgtcg
     5341 tacttggtat tcatgcgtga gtcaaggatt tgggctacat gtttagtaat ctgccgagtc
     5401 tccacaagtt gccgtttaat gaatcctgct ttgtccagtt cgctcagtcc gcccctctca
     5461 gccttagtga gattgtcgaa cttccgctga gtgattagct ttgcgttcag aagctggcgc
     5521 caatagttct tcatcttttt cacgacttcc tctgagggca cgttatctga ctttccccgg
     5581 ttcttgtctg agcgagtcag caccttattg tcaattgagt cgtctttgag gaagctctga
     5641 gggacgatgt ggtccacgtc gtagtctgag agcctgttga tatcaagctc ttggtccaca
     5701 tacatatcgc gtccattttg aaggtagtac agatagagct tctcattctg aagctgggtg
     5761 ttttccaccg ggtgctcttt aaggatctgt gacccgagct ccttaatccc ttcctcaatt
     5821 cttttcatcc gctccctgct gttcttctgt cccttctggg tagtctggtt ttccctggcc
     5881 atctcgatga caatgttctc tggtttatgg cgacccatca ccttgaccag ctcgtccacg
     5941 actttcacgg tctggagaat acccttctta atagcgggtg aaccagccag atttgcgata
     6001 tgctcgtgca gtgagtcacc ttgtccagac acttgggcct tctggatgtc ctccttaaag
     6061 gtcagggaat catcatggat aagttgcatg aagttgcgat tagcgaatcc gtcgctctta
     6121 aggaaatcaa gaatggtctt tccgctctgt ttgtcccgga ttccgttgat gagtttgcgg
     6181 ctaaggcgtc cccatccggt atatcttctc cgtttaagtt gcttcatcac tttatcgtcg
     6241 aacagatggg cgtaggtctt aagcctctcc tcgatcattt cccggtcctc gaacagagtg
     6301 agagtcagga caatgtcctc aaggatgtcc tctttctcct cattgtccag aaaatccttg
     6361 tccttgatga tcttaagcag atcatggtag gtacccaggc ttgcgttgaa ccgatcttcg
     6421 accccgctaa tctccacaga gtcgaagcat tcaattttct tgaaatagtc ctccttcagc
     6481 tgcttcacgg tcacctttct attggtcttg aacagaaggt ccacaattgc cttcttttgt
     6541 tcgcctgaca ggaatgcggg cttcctcatc ccttcggtga catatttcac cttggtcagc
     6601 tcgttgtaca cggtaaagta ctcgtagagc agggaatgtt tcggcaggac cttctcattg
     6661 gggaggttct tatcgaagtt agtcatccgt tcgatgaatg actgagcgct ggctcccttg
     6721 tccaccacct cttcgaagtt ccagggagtg atagtttctt ctgactttct ggtcatccaa
     6781 gcaaaccggc tatttcctct ggcgaggggt cccacgtagt aggggatgcg gaaagtgaga
     6841 attttctcaa tcttctccct gttgtcctta agaaaggggt agaagtcctc ttggcgccga
     6901 aggatggcgt gaagctcccc aagatggatc tggtggggaa tgctcccatt gtcgaaagtg
     6961 cgctgcttcc gcagcagatc ctcccgattc agtttcacga ggagctcttc ggtgccgtcc
     7021 atcttctcaa ggataggttt gataaactta taaaattcct cttgggaggc acctccgtcg
     7081 atgtagccgg cgtatccatt ctttgactgg tcaaagaaga tttccttgta cttctccgga
     7141 agctgctgcc ggacgagtgc cttaagcagg gtcaggtcct ggtggtgctc atcataccgc
     7201 ttaatcattg aagctgaaag aggggccttg gtgatttcag tgttcacgcg gagaatgtca
     7261 gacagcagga tggcgtcaga aagattctta gcagcaagga aaaggtcggc gtattggtca
     7321 ccgatctggg ccaggaggtt gtcgagatcg tcatcatagg tgtccttgga cagttgcagc
     7381 ttggcatcct ctgccaggtc gaaattagac ttgaaattgg gagtcagccc aagtgaaaga
     7441 gcaatcaggt tcccgaacag tccatttttc ttttcacccg gcagctgagc gataagattc
     7501 tcaagtctgc ggctctttga cagccgggct gacagaatgg ccttggcatc gactccgctt
     7561 gcattgatgg ggttctcttc aaagagctgg ttgtaagtct gcaccagctg gatgaacagt
     7621 ttatccacat cgctattgtc aggattgagg tcgccctcga tcaggaagtg gcctctgaac
     7681 ttaatcatgt gggccagggc caggtagata agccggaggt ctgccttgtc ggttgagtcc
     7741 accagctttt tgcgaagatg gtagatggtt gggtatttct cgtgataggc cacctcgtcg
     7801 acgatattac cgaagatggg atgccgttca tgcttcttat cctcttccac gaggaatgat
     7861 tcttccagcc tatggaagaa gctgtcgtcg acctttgcca tttcgttgga gaatatctct
     7921 tgaaggtaac aaatccggtt cttcctcctg gtgtaccgcc gtctagcagt ccgcttgagg
     7981 cgggttgctt cagcggtctc ccctgaatca aagaggagag ctccgatcag atttttcttg
     8041 atggaatgcc ggtcggtgtt cccgagcacc ttgaacttct tagagggcac cttgtactcg
     8101 tcagtgatga cggcccaccc gacgctatta gtcccgatgt ccagtccgat agaatacttc
     8161 ttgtccatgg tggcgagatc taattcgatc catatcagat cttttgtttg tttatgtgtg
     8221 tttattcgaa actaagttct tggtgtttta aaactaaaaa aaagactaac tataaaagta
     8281 gaatttaaga agtttaagaa atagatttac agaattacaa tcaataccta ccgtctttat
     8341 atacttatta gtcaagtagg ggaataattt cagggaactg gtttcaacct tttttttcag
     8401 ctttttccaa atcagagaga gcagaaggta atagaaggtg taagaaaatg agatagatac
     8461 atgcgtgggt caattgcctt gtgtcatcat ttactccagg caggttgcat cactccattg
     8521 aggttgtgcc cgttttttgc ctgtttgtgc ccctgttctc tgtagttgcg ctaagagaat
     8581 ggacctatga actgatggtt ggtgaagaaa acaatatttt ggtgctggga ttcttttttt
     8641 ttctggatgc cagcttaaaa agcgggctcc attatattta gtggatgcca ggaataaact
     8701 gttcacccag acacctacga tgttatatat tctgtgtaac ccgcccccta ttttgggcat
     8761 gtacgggtta cagcagaatt aaaaggctaa ttttttgact aaataaagtt aggaaaatca
     8821 ctactattaa ttatttacgt attctttgaa atggcagtat tgataatgat aaactcgaac
     8881 tgactagtag actgaattcg atatcaagct tatcgatacc gtcgacctcg agggggggcc
     8941 cggtaccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc
     9001 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     9061 aagcataaag tgtaaagcct ggggtgccta atgagtgagg tacggatcga tcctgacagg
     9121 atatattggc gggtaaacct aagagaaaag agcgtttatt agaataatcg gatatttaaa
     9181 agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acagggttcc
     9241 ccagatctgg cgccggccag cgagacgagc aagattggcc gcggtcacac tgcttccggt
     9301 agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga
     9361 accgacgacc gggtcgaatt tgctttcgaa ccccagagtc ccgctcagaa gaactcgtca
     9421 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     9481 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     9541 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     9601 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     9661 tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     9721 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     9781 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     9841 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     9901 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     9961 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc
    10021 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
    10081 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
    10141 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
    10201 cgaaacgatc ctcatcctgt ctcttgatca gatcttgatc ccctgcgcca tcagatcctt
    10261 ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc
    10321 ccagctggca attccggttc gcttgctgtc cataaaaccg cccagtctag ctatcgccat
    10381 gtaagcccac tgcctcgaga ctagttggtg tccaaccggc tcgacggggg cagcgcaagg
    10441 cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg cttcgcaaag
    10501 tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc ttcgccctgc
    10561 gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc gagcggccac
    10621 cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga tggacaggct
    10681 gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc ttcgaccaca
    10741 tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt ttaattttct
    10801 ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg acaccaagtg
    10861 gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac gcgcctggaa
    10921 cgacccaagc ctatgcgagt gggggcagtc gaagggcgaa gcccgcccgc ctgccccccg
    10981 agcctcacgg cggcgagtgc gggggttcca agggggcagc gccaccttgg gcaaggccga
    11041 aggccgcgca gtcgatcaac aagccccgga ggggccactt tttgccggag ggggagccgc
    11101 gccgaaggcg tgggggaacc ccgcaggggt gcccttcttt gggcaccaaa gaactagata
    11161 tagggcgaaa tgcgaaagac ttaaaaatca acaacttaaa aaaggggggt acgcaacagc
    11221 tcattgcggc accccccgca atagctcatt gcgtaggtta aagaaaatct gtaattgact
    11281 gccactttta cgcaacgcat aattgttgtc gcgctgccga aaagttgcag ctgattgcgc
    11341 atggtgccgc aaccgtgcgg cacccctacc gcatggagat aagcatggcc acgcagtcca
    11401 gagaaatcgg cattcaagcc aagaacaagc ccggtcactg ggtgcaaacg gaacgcaaag
    11461 cgcatgaggc gtgggccggg cttattgcga ggaaacccac ggcggcaatg ctgctgcatc
    11521 acctcgtggc gcagatgggc caccagaacg ccgtggtggt cagccagaag acactttcca
    11581 agctcatcgg acgttctttg cggacggtcc aatacgcagt caaggacttg gtggccgagc
    11641 gctggatctc cgtcgtgaag ctcaacggcc ccggcaccgt gtcggcctac gtggtcaatg
    11701 accgcgtggc gtggggccag ccccgcgacc agttgcgcct gtcggtgttc agtgccgccg
    11761 tggtggttga tcacgacgac caggacgaat cgctgttggg gcatggcgac ctgcgccgca
    11821 tcccgaccct gtatccgggc gagcagcaac taccgaccgg ccccggcgag gagccgccca
    11881 gccagcccgg cattccgggc atggaaccag acctgccagc cttgaccgaa acggaggaat
    11941 gggaacggcg cgggcagcag cgcctgccga tgcccgatga gccgtgtttt ctggacgatg
    12001 gcgagccgtt ggagccgccg acacgggtca cgctgccgcg ccggtagcac ttgggttgcg
    12061 cagcaacccg taagtgcgct gttccagact atcggctgta gccg
//
