LOCUS       sequence-449564-       11243 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 11243)
  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 11243)
  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: pYAMTrGCas
FEATURES             Location/Qualifiers
     source          1..11243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    10..513
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     555..573
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(611..633)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     733..752
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     gene            808..2164
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        808..1151
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1152..1961
                     /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(1219..1238)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1967..2164
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(2246..2701)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2383..2404)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2595..2614
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2827..2849
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2841..2858
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2842..2858
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2865..2884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2865..2883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2959..3146)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     3067..3091
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3174..3194)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(5589..7307)
                     /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      7304..7313
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7342..8008)
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     complement(7449..7472)
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(8053..8069)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8054..8070)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(8098..8116)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8137..8153)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8137..8153)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8150..8172)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8161..8177
                     /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(8185..8191,8192..8209,8210..8215))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_feature    8253..8277
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(8544..9338)
                     /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(8655..8674)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     9265..9284
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      9752..10523
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             10524..11186
                     /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 tcacgtgcta taaaaataat tataatttaa attttttaat ataaatatat
       61 aaattaaaaa tagaaagtaa aaaaagaaat taaagaaaaa atagtttttg ttttccgaag
      121 atgtaaaaga ctctaggggg atcgccaaca aatactacct tttatcttgc tcttcctgct
      181 ctcaggtatt aatgccgaat tgtttcatct tgtctgtgta gaagaccaca cacgaaaatc
      241 ctgtgatttt acattttact tatcgttaat cgaatgtata tctatttaat ctgcttttct
      301 tgtctaataa atatatatgt aaagtacgct ttttgttgaa attttttaaa cctttgttta
      361 tttttttttc ttcattccgt aactcttcta ccttctttat ttactttcta aaatccaaat
      421 acaaaacata aaaataaata aacacagagt aaattcccaa attattccat cattaaaaga
      481 tacgaggcgc gtgtaagtta caggcaagcg atccgtccta agaaaccatt attatcatga
      541 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
      601 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
      661 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
      721 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatagt ttagcttgcc
      781 tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct ccttgacagt
      841 cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat
      901 ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat
      961 tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat
     1021 tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc
     1081 ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa
     1141 cataaacaac catgggtaag gaaaagactc acgtttcgag gccgcgatta aattccaaca
     1201 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     1261 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     1321 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     1381 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     1441 ctgcgatccc cggcaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     1501 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     1561 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     1621 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     1681 ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt
     1741 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     1801 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     1861 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     1921 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagtactg acaataaaaa
     1981 gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt
     2041 ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc
     2101 gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat
     2161 actgctgtcg attcgatact aacgccgcca tccagtgtcg aaaacaatac cgcacagatg
     2221 cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg
     2281 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2341 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2401 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2461 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2521 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2581 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2641 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2701 tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     2761 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     2821 ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat
     2881 agggcgaatt ggagctccac cgcggtggcg gccgctctag agaaatgggg agcgatttgc
     2941 aggcatttgc tcggcatgcc ggtagaggtg tggtcaataa gagcgacctc atgctatacc
     3001 tgagaaagca acctgaccta caggaaagag ttactcaaga ataagaattt tcgttttaaa
     3061 acctaagagt cactttaaaa tttgtataca cttatttttt ttataactta tttaataata
     3121 aaaatcataa atcataagaa attcgcctcg agagtctagg atccggaact acctaccttg
     3181 cgctttttct tgggagctcc ctcatcccct ccgagctgtg agaggtcaat ccgagtttcg
     3241 taaagtccgg tgatgctctg gtggatgagg gtggcgtcca gcacttcttt ggtggaggta
     3301 tagcgtttcc ggtcgatggt ggtgtcgaag tacttgaatg cggcaggggc accaagattg
     3361 gtgagggtga acaggtggat gatattctca gcctgctccc gaattggttt gtctctgtgc
     3421 ttattgtatg cgctaaggac tttatcgagg ttggcatcag cgagaatgac gcgtttagaa
     3481 aactcggaaa tctgctcaat gatctcgtcc agataatgct tatgctgctc cacaaacagt
     3541 tgcttttgct cgttgtcctc ggggctcccc ttcagcttct cgtaatggct tgcaaggtag
     3601 aggaaattca cgtacttgga ggggagagcc agctcattac ccttctgaag ttcgccagcg
     3661 gaggcgagca tccgcttccg cccattctcc agttcgaaca ggctatactt tgggagcttg
     3721 atgataaggt ccttcttgac ttccttgtaa cctttggctt cgagaaagtc aattgggttc
     3781 ttctcgaagg agctgcgctc cataatggtg atacccagca gttccttcac gctcttgagc
     3841 ttcttagact ttcctttttc caccttagcc acgaccagga cagaataagc cacagttggt
     3901 gaatcgaatc cgccgtactt cttagggtcc caatccttct tccttgcaat gagcttgtcg
     3961 gagttgcgct tggggaggat tgattcctta gaaaagcctc cggtttgcac ctcggttttc
     4021 ttcacgatgt tcacttgagg catagaaagc actttgcgca cggtagcgaa atctctgccc
     4081 ttgtcccaca cgatttcgcc agtctcccca ttagtctcga tgagcggcct cttccggatc
     4141 tcaccgtttg caagggtgat ttcggtcttg aagaaattca tgatgttgct gtagaagaag
     4201 tactttgcgg tggcctttcc gatctcctgc tcagatttgg caatcatctt gcgcacgtcg
     4261 tagaccttgt agtctccgta cacgaactcg ctttccagct ttgggtactt cttgattagg
     4321 gcggttccca ccacggcatt cagatatgcg tcatgagcat gatgatagtt attgatctcc
     4381 cgcactttgt agaattgaaa gtcctttctg aaatcgctga ccagcttgga cttcagggta
     4441 atcaccttca cctcgcggat aagtttgtcg ttttcgtcgt acttggtatt catgcgtgag
     4501 tcaaggattt gggctacatg tttagtaatc tgccgagtct ccacaagttg ccgtttaatg
     4561 aatcctgctt tgtccagttc gctcagtccg cccctctcag ccttagtgag attgtcgaac
     4621 ttccgctgag tgattagctt tgcgttcaga agctggcgcc aatagttctt catctttttc
     4681 acgacttcct ctgagggcac gttatctgac tttccccggt tcttgtctga gcgagtcagc
     4741 accttattgt caattgagtc gtctttgagg aagctctgag ggacgatgtg gtccacgtcg
     4801 tagtctgaga gcctgttgat atcaagctct tggtccacat acatatcgcg tccattttga
     4861 aggtagtaca gatagagctt ctcattctga agctgggtgt tttccaccgg gtgctcttta
     4921 aggatctgtg acccgagctc cttaatccct tcctcaattc ttttcatccg ctccctgctg
     4981 ttcttctgtc ccttctgggt agtctggttt tccctggcca tctcgatgac aatgttctct
     5041 ggtttatggc gacccatcac cttgaccagc tcgtccacga ctttcacggt ctggagaata
     5101 cccttcttaa tagcgggtga accagccaga tttgcgatat gctcgtgcag tgagtcacct
     5161 tgtccagaca cttgggcctt ctggatgtcc tccttaaagg tcagggaatc atcatggata
     5221 agttgcatga agttgcgatt agcgaatccg tcgctcttaa ggaaatcaag aatggtcttt
     5281 ccgctctgtt tgtcccggat tccgttgatg agtttgcggc taaggcgtcc ccatccggta
     5341 tatcttctcc gtttaagttg cttcatcact ttatcgtcga acagatgggc gtaggtctta
     5401 agcctctcct cgatcatttc ccggtcctcg aacagagtga gagtcaggac aatgtcctca
     5461 aggatgtcct ctttctcctc attgtccaga aaatccttgt ccttgatgat cttaagcaga
     5521 tcatggtagg tacccaggct tgcgttgaac cgatcttcga ccccgctaat ctccacagag
     5581 tcgaagcatt caattttctt gaaatagtcc tccttcagct gcttcacggt cacctttcta
     5641 ttggtcttga acagaaggtc cacaattgcc ttcttttgtt cgcctgacag gaatgcgggc
     5701 ttcctcatcc cttcggtgac atatttcacc ttggtcagct cgttgtacac ggtaaagtac
     5761 tcgtagagca gggaatgttt cggcaggacc ttctcattgg ggaggttctt atcgaagtta
     5821 gtcatccgtt cgatgaatga ctgagcgctg gctcccttgt ccaccacctc ttcgaagttc
     5881 cagggagtga tagtttcttc tgactttctg gtcatccaag caaaccggct atttcctctg
     5941 gcgaggggtc ccacgtagta ggggatgcgg aaagtgagaa ttttctcaat cttctccctg
     6001 ttgtccttaa gaaaggggta gaagtcctct tggcgccgaa ggatggcgtg aagctcccca
     6061 agatggatct ggtggggaat gctcccattg tcgaaagtgc gctgcttccg cagcagatcc
     6121 tcccgattca gtttcacgag gagctcttcg gtgccgtcca tcttctcaag gataggtttg
     6181 ataaacttat aaaattcctc ttgggaggca cctccgtcga tgtagccggc gtatccattc
     6241 tttgactggt caaagaagat ttccttgtac ttctccggaa gctgctgccg gacgagtgcc
     6301 ttaagcaggg tcaggtcctg gtggtgctca tcataccgct taatcattga agctgaaaga
     6361 ggggccttgg tgatttcagt gttcacgcgg agaatgtcag acagcaggat ggcgtcagaa
     6421 agattcttag cagcaaggaa aaggtcggcg tattggtcac cgatctgggc caggaggttg
     6481 tcgagatcgt catcataggt gtccttggac agttgcagct tggcatcctc tgccaggtcg
     6541 aaattagact tgaaattggg agtcagccca agtgaaagag caatcaggtt cccgaacagt
     6601 ccatttttct tttcacccgg cagctgagcg ataagattct caagtctgcg gctctttgac
     6661 agccgggctg acagaatggc cttggcatcg actccgcttg cattgatggg gttctcttca
     6721 aagagctggt tgtaagtctg caccagctgg atgaacagtt tatccacatc gctattgtca
     6781 ggattgaggt cgccctcgat caggaagtgg cctctgaact taatcatgtg ggccagggcc
     6841 aggtagataa gccggaggtc tgccttgtcg gttgagtcca ccagcttttt gcgaagatgg
     6901 tagatggttg ggtatttctc gtgataggcc acctcgtcga cgatattacc gaagatggga
     6961 tgccgttcat gcttcttatc ctcttccacg aggaatgatt cttccagcct atggaagaag
     7021 ctgtcgtcga cctttgccat ttcgttggag aatatctctt gaaggtaaca aatccggttc
     7081 ttcctcctgg tgtaccgccg tctagcagtc cgcttgaggc gggttgcttc agcggtctcc
     7141 cctgaatcaa agaggagagc tccgatcaga tttttcttga tggaatgccg gtcggtgttc
     7201 ccgagcacct tgaacttctt agagggcacc ttgtactcgt cagtgatgac ggcccacccg
     7261 acgctattag tcccgatgtc cagtccgata gaatacttct tgtccatggt ggcgagatct
     7321 aattcgatcc atatcagatc ttttgtttgt ttatgtgtgt ttattcgaaa ctaagttctt
     7381 ggtgttttaa aactaaaaaa aagactaact ataaaagtag aatttaagaa gtttaagaaa
     7441 tagatttaca gaattacaat caatacctac cgtctttata tacttattag tcaagtaggg
     7501 gaataatttc agggaactgg tttcaacctt ttttttcagc tttttccaaa tcagagagag
     7561 cagaaggtaa tagaaggtgt aagaaaatga gatagataca tgcgtgggtc aattgccttg
     7621 tgtcatcatt tactccaggc aggttgcatc actccattga ggttgtgccc gttttttgcc
     7681 tgtttgtgcc cctgttctct gtagttgcgc taagagaatg gacctatgaa ctgatggttg
     7741 gtgaagaaaa caatattttg gtgctgggat tctttttttt tctggatgcc agcttaaaaa
     7801 gcgggctcca ttatatttag tggatgccag gaataaactg ttcacccaga cacctacgat
     7861 gttatatatt ctgtgtaacc cgccccctat tttgggcatg tacgggttac agcagaatta
     7921 aaaggctaat tttttgacta aataaagtta ggaaaatcac tactattaat tatttacgta
     7981 ttctttgaaa tggcagtatt gataatgata aactcgaact gactagtaga ctgaattcga
     8041 tatcaagctt atcgataccg tcgacctcga gggggggccc ggtaccagct tttgttccct
     8101 ttagtgaggg ttaatttcga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     8161 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     8221 gggtgcctaa tgagtgaggt acggatcgat cctgacagga tatattggcg ggtaaaccta
     8281 agagaaaaga gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc
     8341 gttcgtccat ttgtatgtgc atgccaacca cagggttccc cagatctggc gccggccagc
     8401 gagacgagca agattggccg cggtcacact gcttccggta gtcaataaac cggtaaacca
     8461 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt
     8521 gctttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg
     8581 cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg
     8641 ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca
     8701 cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc
     8761 aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc
     8821 ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg
     8881 acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg
     8941 aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat
     9001 actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat
     9061 agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc
     9121 gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac
     9181 aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca
     9241 tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg
     9301 gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc
     9361 tcttgatcag atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag
     9421 tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa ttccggttcg
     9481 cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg taagcccact gcctcgagac
     9541 tagttggtgt ccaaccggct cgacgggggc agcgcaaggc ggtgcctccg gcgggccact
     9601 caatgcttga gtatactcac tagactttgc ttcgcaaagt cgtgaccgcc tacggcggct
     9661 gcggcgccct acgggcttgc tctccgggct tcgccctgcg cggtcgctgc gctcccttgc
     9721 cagcccgtgg atatgtggac gatggccgcg agcggccacc ggctggctcg cttcgctcgg
     9781 cccgtggaca accctgctgg acaagctgat ggacaggctg cgcctgccca cgagcttgac
     9841 cacagggatt gcccaccggc tacccagcct tcgaccacat acccaccggc tccaactgcg
     9901 cggcctgcgg ccttgcccca tcaatttttt taattttctc tggggaaaag cctccggcct
     9961 gcggcctgcg cgcttcgctt gccggttgga caccaagtgg aaggcgggtc aaggctcgcg
    10021 cagcgaccgc gcagcggctt ggccttgacg cgcctggaac gacccaagcc tatgcgagtg
    10081 ggggcagtcg aagggcgaag cccgcccgcc tgccccccga gcctcacggc ggcgagtgcg
    10141 ggggttccaa gggggcagcg ccaccttggg caaggccgaa ggccgcgcag tcgatcaaca
    10201 agccccggag gggccacttt ttgccggagg gggagccgcg ccgaaggcgt gggggaaccc
    10261 cgcaggggtg cccttctttg ggcaccaaag aactagatat agggcgaaat gcgaaagact
    10321 taaaaatcaa caacttaaaa aaggggggta cgcaacagct cattgcggca ccccccgcaa
    10381 tagctcattg cgtaggttaa agaaaatctg taattgactg ccacttttac gcaacgcata
    10441 attgttgtcg cgctgccgaa aagttgcagc tgattgcgca tggtgccgca accgtgcggc
    10501 acccctaccg catggagata agcatggcca cgcagtccag agaaatcggc attcaagcca
    10561 agaacaagcc cggtcactgg gtgcaaacgg aacgcaaagc gcatgaggcg tgggccgggc
    10621 ttattgcgag gaaacccacg gcggcaatgc tgctgcatca cctcgtggcg cagatgggcc
    10681 accagaacgc cgtggtggtc agccagaaga cactttccaa gctcatcgga cgttctttgc
    10741 ggacggtcca atacgcagtc aaggacttgg tggccgagcg ctggatctcc gtcgtgaagc
    10801 tcaacggccc cggcaccgtg tcggcctacg tggtcaatga ccgcgtggcg tggggccagc
    10861 cccgcgacca gttgcgcctg tcggtgttca gtgccgccgt ggtggttgat cacgacgacc
    10921 aggacgaatc gctgttgggg catggcgacc tgcgccgcat cccgaccctg tatccgggcg
    10981 agcagcaact accgaccggc cccggcgagg agccgcccag ccagcccggc attccgggca
    11041 tggaaccaga cctgccagcc ttgaccgaaa cggaggaatg ggaacggcgc gggcagcagc
    11101 gcctgccgat gcccgatgag ccgtgttttc tggacgatgg cgagccgttg gagccgccga
    11161 cacgggtcac gctgccgcgc cggtagcact tgggttgcgc agcaacccgt aagtgcgctg
    11221 ttccagacta tcggctgtag ccg
//
