LOCUS       sequence-372326-        8757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for yeast expression of Cas9; contains a golden gate cloning
            site to additionally express a gRNA for a target sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8757)
  AUTHORS   Aguilar RR, Shen ZJ, Tyler JK
  TITLE     A Simple, Improved Method for Scarless Genome Editing of Budding 
            Yeast Using CRISPR-Cas9.
  JOURNAL   Methods Protoc. 2022 Oct 4;5(5):79. doi: 10.3390/mps5050079.
  PUBMED    36287051
REFERENCE   2  (bases 1 to 8757)
  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: "Methods 
            Protoc."; date: "2022-10-4"; volume: "5(5):79. doi"; pages: " 
            10.3390/mps5050079"
COMMENT     Sequence Label: pCASB
FEATURES             Location/Qualifiers
     source          1..8757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(749..796)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_RNA        1183..1258
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     gene            1392..2748
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1392..1735
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1736..2545
                     /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(1803..1822)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2551..2748
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             3511..7614
                     /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             3511..7614
                     /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             7624..7644
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7660..7683
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      7687..7876
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(7933..8521)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8013..8032)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      join(8543..8757,1..768)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg
       61 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
      121 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
      181 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
      241 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
      301 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
      361 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
      421 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
      481 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
      541 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
      601 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
      661 aatattttag tagcccgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcttca
      721 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagag aataggaact
      781 tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc
      841 tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata
      901 tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtatat gtgttatgta
      961 gtatactctt tcttcaacaa ttaaatactc tcggtagcca agttggttta aggcgcaaga
     1021 ctgtaattta tcactacgaa atcttgagat cgggcgttcg actcgccccc gggagagatg
     1081 gccggcatgg tcccagcctc ctcgctggcg ccggctgggc aacaccttcg ggtggcgaat
     1141 gggactttag agacccataa taacgtccaa taaatggtct cagttttaga gctagaaata
     1201 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     1261 tttttatttt ttgtcactat tgttatgtaa aatgccacct ctgacagtat ggaacgcaaa
     1321 cttctgtcta gtggatatag gtctagagat ctgtttagct tgcctcgtcc ccgccgggtc
     1381 acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct
     1441 caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat
     1501 ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc
     1561 agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc
     1621 ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc
     1681 cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa caaccatggg
     1741 taaggaaaag actcacgttt cgaggccgcg attaaattcc aacatggatg ctgatttata
     1801 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta
     1861 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     1921 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     1981 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggcaa
     2041 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2101 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2161 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     2221 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     2281 gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     2341 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     2401 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     2461 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     2521 tttgatgctc gatgagtttt tctaatcagt actgacaata aaaagattct tgttttcaag
     2581 aacttgtcat ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc
     2641 gtgatttata ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag
     2701 aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga
     2761 tactaacgcc gccatccagt gtcgaaaacg agctctcgag aacccttaat agtcgaacaa
     2821 gaagcaggca aagtttagag cactgcccct ccgcactcaa aaaagaaaaa actaggagga
     2881 aaataaaatt ctcaaccaca caaacacata aacacataca aatacaaata caagcttatt
     2941 tacttgacat cgcgcgatct tccactattc agcgccgtcc gccctctctc gtgttttttg
     3001 tttacgcgac aactatgcga aatccggagc aacgggcaac cgtttgggga aagaccacac
     3061 ccacgcgcga tcgccatggc aacgaggtcg cacacgcccc acacccagac ctccctgcga
     3121 gcgggcatgg gtacaatgtc cccgttgcca cagacaccac ttcgtagcac agcgcagagc
     3181 gtagcgtgtt gttgctgctg acaaaagaaa atttttctta gcaaagcaaa ggaggggaag
     3241 cacgggcaga tagcaccgta ccataccctt ggaaactcga aatgaacgaa gcaggaaatg
     3301 agagaatgag agttttgtag gtatatatag cggtagtgtt tgcgcgttac catcatcttc
     3361 tggatctatc tattgttctt ttcctcatca ctttcccctt tttcgctctt cttcttgtct
     3421 tttatttctt tctttttttt aattgttccc tcgattggct atctaccaaa gaatccaaac
     3481 ttaatacacg tatttatttg tccaattacc atggataaga aatactcaat aggcttagat
     3541 atcggcacaa atagcgtcgg atgggcggtg atcactgatg aatataaggt tccgtctaaa
     3601 aagttcaagg ttctgggaaa tacagaccgc cacagtatca aaaaaaatct tataggggct
     3661 cttttatttg acagtggaga gacagcggaa gcgactcgtc tcaaacggac agctcgtaga
     3721 aggtatacac gtcggaagaa tcgtatttgt tatctacagg agattttttc aaatgagatg
     3781 gcgaaagtag atgatagttt ctttcatcga cttgaagagt cttttttggt ggaagaagac
     3841 aagaagcatg aacgtcatcc tatttttgga aatatagtag atgaagttgc ttatcatgag
     3901 aaatatccaa ctatctatca tctgcgaaaa aaattggtag attctactga taaagcggat
     3961 ttgcgcttaa tctatttggc cttagcgcat atgattaagt ttcgtggtca ttttttgatt
     4021 gagggagatt taaatcctga taatagtgat gtggacaaac tatttatcca gttggtacaa
     4081 acctacaatc aattatttga agaaaaccct attaacgcaa gtggagtaga tgctaaagcg
     4141 attctttctg cacgattgag taaatcaaga cgattagaaa atctcattgc tcagctcccc
     4201 ggtgagaaga aaaatggctt atttgggaat ctcattgctt tgtcattggg tttgacccct
     4261 aattttaaat caaattttga tttggcagaa gatgctaaat tacagctttc aaaagatact
     4321 tacgatgatg atttagataa tttattggcg caaattggag atcaatatgc tgatttgttt
     4381 ttggcagcta agaatttatc agatgctatt ttactttcag atatcctaag agtaaatact
     4441 gaaataacta aggctcccct atcagcttca atgattaaac gctacgatga acatcatcaa
     4501 gacttgactc ttttaaaagc tttagttcga caacaacttc cagaaaagta taaagaaatc
     4561 ttttttgatc aatcaaaaaa cggatatgca ggttatattg atgggggagc tagccaagaa
     4621 gaattttata aatttatcaa accaatttta gaaaaaatgg atggtactga ggaattattg
     4681 gtgaaactaa atcgtgaaga tttgctgcgc aagcaacgga cctttgacaa cggctctatt
     4741 ccccatcaaa ttcacttggg tgagctgcat gctattttga gaagacaaga agacttttat
     4801 ccatttttaa aagacaatcg tgagaagatt gaaaaaatct tgacttttcg aattccttat
     4861 tatgttggtc cattggcgcg tggcaatagt cgttttgcat ggatgactcg gaagtctgaa
     4921 gaaacaatta ccccatggaa ttttgaagaa gttgtcgata aaggtgcttc agctcaatca
     4981 tttattgaac gcatgacaaa ctttgataaa aatcttccaa atgaaaaagt actaccaaaa
     5041 catagtttgc tttatgagta ttttacggtt tataacgaat tgacaaaggt caaatatgtt
     5101 actgaaggaa tgcgaaaacc agcatttctt tcaggtgaac agaagaaagc cattgttgat
     5161 ttactcttca aaacaaatcg aaaagtaacc gttaagcaat taaaagaaga ttatttcaaa
     5221 aaaatagaat gttttgatag tgttgaaatt tcaggagttg aagatagatt taatgcttca
     5281 ttaggtacct accatgattt gctaaaaatt attaaagata aagatttttt ggataatgaa
     5341 gaaaatgaag atatcttaga ggatattgtt ttaacattga ccttatttga agatagggag
     5401 atgattgagg aaagacttaa aacatatgct cacctctttg atgataaggt gatgaaacag
     5461 cttaaacgtc gccgttatac tggttgggga cgtttgtctc gaaaattgat taatggtatt
     5521 agggataagc aatctggcaa aacaatatta gattttttga aatcagatgg ttttgccaat
     5581 cgcaatttta tgcagctgat ccatgatgat agtttgacat ttaaagaaga cattcaaaaa
     5641 gcacaagtgt ctggacaagg cgatagttta catgaacata ttgcaaattt agctggtagc
     5701 cctgctatta aaaaaggtat tttacagact gtaaaagttg ttgatgaatt ggtcaaagta
     5761 atggggcggc ataagccaga aaatatcgtt attgaaatgg cacgtgaaaa tcagacaact
     5821 caaaagggcc agaaaaattc gcgagagcgt atgaaacgaa tcgaagaagg tatcaaagaa
     5881 ttaggaagtc agattcttaa agagcatcct gttgaaaata ctcaattgca aaatgaaaag
     5941 ctctatctct attatctcca aaatggaaga gacatgtatg tggaccaaga attagatatt
     6001 aatcgtttaa gtgattatga tgtcgatcac attgttccac aaagtttcct taaagacgat
     6061 tcaatagaca ataaggtctt aacgcgttct gataaaaatc gtggtaaatc ggataacgtt
     6121 ccaagtgaag aagtagtcaa aaagatgaaa aactattgga gacaacttct aaacgccaag
     6181 ttaatcactc aacgtaagtt tgataattta acgaaagctg aacgtggagg tttgagtgaa
     6241 cttgataaag ctggttttat caaacgccaa ttggttgaaa ctcgccaaat cactaagcat
     6301 gtggcacaaa ttttggatag tcgcatgaat actaaatacg atgaaaatga taaacttatt
     6361 cgagaggtta aagtgattac cttaaaatct aaattagttt ctgacttccg aaaagatttc
     6421 caattctata aagtacgtga gattaacaat taccatcatg cccatgatgc gtatctaaat
     6481 gccgtcgttg gaactgcttt gattaagaaa tatccaaaac ttgaatcgga gtttgtctat
     6541 ggtgattata aagtttatga tgttcgtaaa atgattgcta agtctgagca agaaataggc
     6601 aaagcaaccg caaaatattt cttttactct aatatcatga acttcttcaa aacagaaatt
     6661 acacttgcaa atggagagat tcgcaaacgc cctctaatcg aaactaatgg ggaaactgga
     6721 gaaattgtct gggataaagg gcgagatttt gccacagtgc gcaaagtatt gtccatgccc
     6781 caagtcaata ttgtcaagaa aacagaagta cagacaggcg gattctccaa ggagtcaatt
     6841 ttaccaaaaa gaaattcgga caagcttatt gctcgtaaaa aagactggga tccaaaaaaa
     6901 tatggtggtt ttgatagtcc aacggtagct tattcagtcc tagtggttgc taaggtggaa
     6961 aaagggaaat cgaagaagtt aaaatccgtt aaagagttac tagggatcac aattatggaa
     7021 agaagttcct ttgaaaaaaa tccgattgac tttttagaag ctaaaggata taaggaagtt
     7081 aaaaaagact taatcattaa actacctaaa tatagtcttt ttgagttaga aaacggtcgt
     7141 aaacggatgc tggctagtgc cggagaatta caaaaaggaa atgagctggc tctgccaagc
     7201 aaatatgtga attttttata tttagctagt cattatgaaa agttgaaggg tagtccagaa
     7261 gataacgaac aaaaacaatt gtttgtggag cagcataagc attatttaga tgagattatt
     7321 gagcaaatca gtgaattttc taagcgtgtt attttagcag atgccaattt agataaagtt
     7381 cttagtgcat ataacaaaca tagagacaaa ccaatacgtg aacaagcaga aaatattatt
     7441 catttattta cgttgacgaa tcttggagct cccgctgctt ttaaatattt tgatacaaca
     7501 attgatcgta aacgatatac gtctacaaaa gaagttttag atgccactct tatccatcaa
     7561 tccatcactg gtctttatga aacacgcatt gatttgagtc agctaggagg tgacggtgga
     7621 gggccaaaaa agaaaagaaa agttgaagat gcttctgctc atcatcacca tcaccatcac
     7681 cattaaatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     7741 ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt
     7801 ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt
     7861 tttgggacgc tcgaagtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7921 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7981 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     8041 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     8101 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     8161 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     8221 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     8281 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     8341 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     8401 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     8461 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     8521 agaagatcct ttgatctttt ctaccgaagc atctgtgctt cattttgtag aacaaaaatg
     8581 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     8641 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     8701 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttaca
//
