LOCUS       sequence-386150-       10373 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allows the expression of Cas9 in Saccharomyces cerevisiae and 
            contains a gRNA expression unit with a mRFP1 placeholder for 
            user-defined gRNA insertion using BsaI-mediated Golden Gate 
            Assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10373)
  AUTHORS   Maestroni L, Butti P, Senatore VG, Branduardi P
  TITLE     pCEC-red: a new vector for easier and faster CRISPR-Cas9 genome 
            editing in Saccharomyces cerevisiae.
  JOURNAL   FEMS Yeast Res. 2023 Jan 4;23:foad002. doi: 10.1093/femsyr/foad002.
  PUBMED    36640150
REFERENCE   2  (bases 1 to 10373)
  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: "FEMS Yeast 
            Res."; date: "2023-01-4"; volume: "23:foad002. doi"; pages: " 
            10.1093/femsyr/foad002"
COMMENT     Sequence Label: pCEC-red
FEATURES             Location/Qualifiers
     source          1..10373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4101..4121
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      4149..4336
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4204..4228)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     terminator      complement(4454..4473)
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     terminator      complement(4555..4582)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(4598..4669)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4714..5391)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             5398..5409
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    5427..5443
                     /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(5451..5457,5458..5475,5476..5481))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5496..5517
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        complement(5531..5799)
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     primer_bind     complement(5949..5966)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6120..6708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6200..6219)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      6802..8144
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(7765..7812)
                     /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)."
     primer_bind     8185..8203
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     gene            8214..9570
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        8214..8557
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             8558..9367
                     /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(8625..8644)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      9373..9570
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     9598..9614
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     9599..9615
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9660..10326
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     10196..10219
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     regulatory      10355..10364
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(10361..10373,1..4091)
                     /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             join(10361..10373,1..4091)
                     /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"
ORIGIN
        1 attctatcgg actggacatc gggactaata gcgtcgggtg ggccgtcatc actgacgagt
       61 acaaggtgcc ctctaagaag ttcaaggtgc tcgggaacac cgaccggcat tccatcaaga
      121 aaaatctgat cggagctctc ctctttgatt caggggaaac cgctgaagca acccgcctca
      181 agcggactgc tagacggcgg tacaccagga ggaagaaccg gatttgttac cttcaagaga
      241 tattctccaa cgaaatggca aaggtcgacg acagcttctt ccataggctg gaagaatcat
      301 tcctcgtgga agaggataag aagcatgaac ggcatcccat cttcggtaat atcgtcgacg
      361 aggtggccta tcacgagaaa tacccaacca tctaccatct tcgcaaaaag ctggtggact
      421 caaccgacaa ggcagacctc cggcttatct acctggccct ggcccacatg attaagttca
      481 gaggccactt cctgatcgag ggcgacctca atcctgacaa tagcgatgtg gataaactgt
      541 tcatccagct ggtgcagact tacaaccagc tctttgaaga gaaccccatc aatgcaagcg
      601 gagtcgatgc caaggccatt ctgtcagccc ggctgtcaaa gagccgcaga cttgagaatc
      661 ttatcgctca gctgccgggt gaaaagaaaa atggactgtt cgggaacctg attgctcttt
      721 cacttgggct gactcccaat ttcaagtcta atttcgacct ggcagaggat gccaagctgc
      781 aactgtccaa ggacacctat gatgacgatc tcgacaacct cctggcccag atcggtgacc
      841 aatacgccga ccttttcctt gctgctaaga atctttctga cgccatcctg ctgtctgaca
      901 ttctccgcgt gaacactgaa atcaccaagg cccctctttc agcttcaatg attaagcggt
      961 atgatgagca ccaccaggac ctgaccctgc ttaaggcact cgtccggcag cagcttccgg
     1021 agaagtacaa ggaaatcttc tttgaccagt caaagaatgg atacgccggc tacatcgacg
     1081 gaggtgcctc ccaagaggaa ttttataagt ttatcaaacc tatccttgag aagatggacg
     1141 gcaccgaaga gctcctcgtg aaactgaatc gggaggatct gctgcggaag cagcgcactt
     1201 tcgacaatgg gagcattccc caccagatcc atcttgggga gcttcacgcc atccttcggc
     1261 gccaagagga cttctacccc tttcttaagg acaacaggga gaagattgag aaaattctca
     1321 ctttccgcat cccctactac gtgggacccc tcgccagagg aaatagccgg tttgcttgga
     1381 tgaccagaaa gtcagaagaa actatcactc cctggaactt cgaagaggtg gtggacaagg
     1441 gagccagcgc tcagtcattc atcgaacgga tgactaactt cgataagaac ctccccaatg
     1501 agaaggtcct gccgaaacat tccctgctct acgagtactt taccgtgtac aacgagctga
     1561 ccaaggtgaa atatgtcacc gaagggatga ggaagcccgc attcctgtca ggcgaacaaa
     1621 agaaggcaat tgtggacctt ctgttcaaga ccaatagaaa ggtgaccgtg aagcagctga
     1681 aggaggacta tttcaagaaa attgaatgct tcgactctgt ggagattagc ggggtcgaag
     1741 atcggttcaa cgcaagcctg ggtacctacc atgatctgct taagatcatc aaggacaagg
     1801 attttctgga caatgaggag aatgaggaca tccttgagga cattgtcctg actctcactc
     1861 tgttcgagga ccgggaaatg atcgaggaga ggcttaagac ctacgcccat ctgttcgacg
     1921 ataaagtgat gaagcaactt aaacggagaa gatataccgg atggggacgc cttagccgca
     1981 aactcatcaa cggaatccgg gacaaacaga gcggaaagac cattcttgat ttccttaaga
     2041 gcgacggatt cgctaatcgc aacttcatgc aacttatcca tgatgattcc ctgaccttta
     2101 aggaggacat ccagaaggcc caagtgtctg gacaaggtga ctcactgcac gagcatatcg
     2161 caaatctggc tggttcaccc gctattaaga agggtattct ccagaccgtg aaagtcgtgg
     2221 acgagctggt caaggtgatg ggtcgccata aaccagagaa cattgtcatc gagatggcca
     2281 gggaaaacca gactacccag aagggacaga agaacagcag ggagcggatg aaaagaattg
     2341 aggaagggat taaggagctc gggtcacaga tccttaaaga gcacccggtg gaaaacaccc
     2401 agcttcagaa tgagaagctc tatctgtact accttcaaaa tggacgcgat atgtatgtgg
     2461 accaagagct tgatatcaac aggctctcag actacgacgt ggaccacatc gtccctcaga
     2521 gcttcctcaa agacgactca attgacaata aggtgctgac tcgctcagac aagaaccggg
     2581 gaaagtcaga taacgtgccc tcagaggaag tcgtgaaaaa gatgaagaac tattggcgcc
     2641 agcttctgaa cgcaaagcta atcactcagc ggaagttcga caatctcact aaggctgaga
     2701 ggggcggact gagcgaactg gacaaagcag gattcattaa acggcaactt gtggagactc
     2761 ggcagattac taaacatgta gcccaaatcc ttgactcacg catgaatacc aagtacgacg
     2821 aaaacgacaa acttatccgc gaggtgaagg tgattaccct gaagtccaag ctggtcagcg
     2881 atttcagaaa ggactttcaa ttctacaaag tgcgggagat caataactat catcatgctc
     2941 atgacgcata tctgaatgcc gtggtgggaa ccgccctaat caagaagtac ccaaagctgg
     3001 aaagcgagtt cgtgtacgga gactacaagg tctacgacgt gcgcaagatg attgccaaat
     3061 ctgagcagga gatcggaaag gccaccgcaa agtacttctt ctacagcaac atcatgaatt
     3121 tcttcaagac cgaaatcacc cttgcaaacg gtgagatccg gaagaggccg ctcatcgaga
     3181 ctaatgggga gactggcgaa atcgtgtggg acaagggcag agatttcgct accgtgcgca
     3241 aagtgctttc tatgcctcaa gtgaacatcg tgaagaaaac cgaggtgcaa accggaggct
     3301 tttctaagga atcaatcctc cccaagcgca actccgacaa gctcattgca aggaagaagg
     3361 attgggaccc taagaagtac ggcggattcg attcaccaac tgtggcttat tctgtcctgg
     3421 tcgtggctaa ggtggaaaaa ggaaagtcta agaagctcaa gagcgtgaag gaactgctgg
     3481 gtatcaccat tatggagcgc agctccttcg agaagaaccc aattgacttt ctcgaagcca
     3541 aaggttacaa ggaagtcaag aaggacctta tcatcaagct cccaaagtat agcctgttcg
     3601 aactggagaa tgggcggaag cggatgctcg cctccgctgg cgaacttcag aagggtaatg
     3661 agctggctct cccctccaag tacgtgaatt tcctctacct tgcaagccat tacgagaagc
     3721 tgaaggggag ccccgaggac aacgagcaaa agcaactgtt tgtggagcag cataagcatt
     3781 atctggacga gatcattgag cagatttccg agttttctaa acgcgtcatt ctcgctgatg
     3841 ccaacctcga taaagtcctt agcgcataca ataagcacag agacaaacca attcgggagc
     3901 aggctgagaa tatcatccac ctgttcaccc tcaccaatct tggtgcccct gccgcattca
     3961 agtacttcga caccaccatc gaccggaaac gctatacctc caccaaagaa gtgctggacg
     4021 ccaccctcat ccaccagagc atcaccggac tttacgaaac tcggattgac ctctcacagc
     4081 tcggagggga tgagggagct cccaagaaaa agcgcaaggt aggtagttcc ggatcctaga
     4141 ctctcgaggc gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa
     4201 taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga
     4261 gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc gctcttattg
     4321 accacacctc taccggcatg ccgagcaaat gcctgcaaat cgctccccat ttctctagag
     4381 cggccgtggt atcgtttaga ttggcaatta cagtgtctta gctcacatgc ttataactaa
     4441 ttacatgact cgaagacata aaaaacaaaa aaagcaccac cgactcggtg ccactttttc
     4501 aagttgataa cggactagcc ttattttaac ttgctatttc gagacctata taaacgcaga
     4561 aaggcccacc cgaaggtgag ccagtgtgac tctagtagag agcgttcacc gacaaacaac
     4621 agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggctc
     4681 tagtagcgat ctacactagc actatcagcg ttattaagca ccggtggagt gacgaccttc
     4741 agcacgttcg tactgttcaa cgatggtgta gtcttcgttg tgggaggtga tgtccagttt
     4801 gatgtcggtt ttgtaagcac ccggcagctg aaccggtttt ttagccatgt aggtggtttt
     4861 aacttcagcg tcgtagtgac caccgtcttt cagtttcaga cgcattttga tttcaccttt
     4921 cagagcaccg tcttccgggt acatacgttc ggtggaagct tcccaaccca tggttttttt
     4981 ctgcataacc ggaccgtcgg acgggaagtt ggtaccacgc agtttaactt tgtagatgaa
     5041 ctcaccgtct tgcagggagg agtcctgggt aacggtaaca acaccaccgt cttcgaagtt
     5101 cataacacgt tcccatttga aaccttccgg gaaggacagt ttcaggtagt ccgggatgtc
     5161 agccgggtgt ttaacgtaag ctttggaacc gtactggaac tgcggggaca ggatgtccca
     5221 agcgaacggc agcggaccac ctttggtaac tttcagttta gcggtctggg taccttcgta
     5281 cggacgacct tcaccttcac cttcgatttc gaactcgtga ccgttaacgg aaccttccat
     5341 acgaactttg aaacgcatga actctttgat aacgtcttcg gaggaagcca tctagtattt
     5401 ctcctctttc tctagtatgt gtgaaattgt tatccgctca caattccaca caacatacga
     5461 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5521 gcgggtctct gatcatttat ctttcactgc ggagaagttt cgaacgccga aacatgcgca
     5581 ccaactttca cttctacagc gtttgaccaa aatcttttga acagaacatt gtagggtgtg
     5641 aaaaaatgcg cacctttacc gctagcccaa gagggcacta caaaatctag agttgtactt
     5701 caaacgtaca tgtaatcacc ttgtatatac tcgaaagaaa acatcaagtt tctgtataaa
     5761 tatgagtgaa agcataatca tacattatct tttcaaagat tgtccgcggt ggagctccag
     5821 cttttgttac taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5881 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5941 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6001 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6061 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6121 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6181 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6241 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6301 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6361 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6421 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6481 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6541 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6601 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6661 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6721 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gcagacacat
     6781 ttccccgaaa agtgccacct gaacgaagca tctgtgcttc attttgtaga acaaaaatgc
     6841 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     6901 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
     6961 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     7021 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
     7081 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
     7141 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
     7201 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
     7261 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
     7321 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
     7381 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
     7441 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
     7501 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
     7561 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
     7621 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
     7681 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc
     7741 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagagaata ggaacttcgg
     7801 aataggaact tcaaagcgtt tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg
     7861 cacatacagc tcactgttca cgtcgcacct atatctgcgt gttgcctgta tatatatata
     7921 catgagaaga acggcatagt gcgtgtttat gcttaaatgc gtacttatat gcgtctattt
     7981 atgtaggatg aaaggtagtc tagtacctcc tgtgatatta tcccattcca tgcggggtat
     8041 cgtatgcttc cttcagcact accctttagc tgttctatat gctgccactc ctcaattgga
     8101 ttagtctcat ccttcaatgc tatcatttcc tttgatattg gatcatacta agaaaccatt
     8161 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg agcgacatgg
     8221 aggcccagaa taccctcctt gacagtcttg acgtgcgcag ctcaggggca tgatgtgact
     8281 gtcgcccgta catttagccc atacatcccc atgtataatc atttgcatcc atacattttg
     8341 atggccgcac ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc gagcagggaa
     8401 acgctcccct cacagacgcg ttgaattgtc cccacgccgc gcccctgtag agaaatataa
     8461 aaggttagga tttgccactg aggttcttct ttcatatact tccttttaaa atcttgctag
     8521 gatacagttc tcacatcaca tccgaacata aacaaccatg ggtaaggaaa agactcacgt
     8581 ttcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     8641 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     8701 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     8761 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     8821 tcctgatgat gcatggttac tcaccactgc gatccccggc aaaacagcat tccaggtatt
     8881 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     8941 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcggctcgc
     9001 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     9061 taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc
     9121 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     9181 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     9241 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     9301 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     9361 tttctaatca gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag
     9421 tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt
     9481 cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc
     9541 aatcgtatgt gaatgctggt cgctatactg tgcccgaatt gggtaccggg ccccccctcg
     9601 aggtcgacgg tatcgataag cttgatatcg aattcagtct actagtcagt tcgagtttat
     9661 cattatcaat actgccattt caaagaatac gtaaataatt aatagtagtg attttcctaa
     9721 ctttatttag tcaaaaaatt agccttttaa ttctgctgta acccgtacat gcccaaaata
     9781 gggggcgggt tacacagaat atataacatc gtaggtgtct gggtgaacag tttattcctg
     9841 gcatccacta aatataatgg agcccgcttt ttaagctggc atccagaaaa aaaaagaatc
     9901 ccagcaccaa aatattgttt tcttcaccaa ccatcagttc ataggtccat tctcttagcg
     9961 caactacaga gaacaggggc acaaacaggc aaaaaacggg cacaacctca atggagtgat
    10021 gcaacctgcc tggagtaaat gatgacacaa ggcaattgac ccacgcatgt atctatctca
    10081 ttttcttaca ccttctatta ccttctgctc tctctgattt ggaaaaagct gaaaaaaaag
    10141 gttgaaacca gttccctgaa attattcccc tacttgacta ataagtatat aaagacggta
    10201 ggtattgatt gtaattctgt aaatctattt cttaaacttc ttaaattcta cttttatagt
    10261 tagtcttttt tttagtttta aaacaccaag aacttagttt cgaataaaca cacataaaca
    10321 aacaaaagat ctgatatgga tcgaattaga tctcgccacc atggacaaga agt
//
