LOCUS       sequence-459849-       11484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast CEN plasmid for galactose-inducible expression of spCas9 and 
            Ec86 reverse transcriptase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11484)
  AUTHORS   Stuecker TN, Hood SE, Molina Pineda J, Lenaduwe S, Winter J, Sadhu 
            MJ, Lewis JA
  TITLE     Improved vectors for retron-mediated CRISPR-Cas9 genome editing in 
            Saccharomyces cerevisiae.
  JOURNAL   G3 (Bethesda). 2025 Aug 4:jkaf175. doi: 10.1093/g3journal/jkaf175.
  PUBMED    40758833
REFERENCE   2  (bases 1 to 11484)
  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: "G3 
            (Bethesda). 2025 Aug 4:jkaf175. doi: 10.1093/g3journal/jkaf175."
COMMENT     Sequence Label: pCas9-EcRT-GAL-Kan
FEATURES             Location/Qualifiers
     source          1..11484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     7..23
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7..23
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        44..62
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     gene            190..1546
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        190..533
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             534..1343
                     /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(601..620)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1349..1546
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1662..1681)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1663..1681)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1688..1705)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1688..1704)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1697..1719)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1845..2300
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1932..1951)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2142..2163
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3605..3625)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        3847..4288
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    3847..3964
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4259..4282
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             4321..8424
                     /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             4321..8424
                     /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             4321..8424
                     /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             8437..8457
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(8489..8507)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      8519..8708
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     8722..8744
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8782..8800)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    8841..9344
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        9381..9485
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9486..9554,9555..10346)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9704..9723)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10517..11105
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11006..11025
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11259..11276
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11393..11414
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11429..11434,11435..11452,11453..11459)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11467..11483
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     join(11472..11484,1..10)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 ttcacacagg aaacagctat gaccatgatt acgccaagct cgaaattaac cctcactaaa
       61 gggaacaaaa gctgctagag cggccgccag ctgaagcttc gtacgctgca ggtcgacgga
      121 tccccgggtt aattaaggcg cgccagatct gtttagcttg cctcgtcccc gccgggtcac
      181 ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca
      241 ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt
      301 gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcgg
      361 acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc
      421 ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct
      481 tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca accatgggta
      541 aggaaaagac tcacgtttcg aggccgcgat taaattccaa catggatgct gatttatatg
      601 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg
      661 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
      721 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
      781 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggcaaaa
      841 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
      901 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
      961 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     1021 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc
     1081 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     1141 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     1201 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     1261 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     1321 tgatgctcga tgagtttttc taatcagtac tgacaataaa aagattcttg ttttcaagaa
     1381 cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt
     1441 gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa
     1501 agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata
     1561 ctaacgccgc catccagtgt cgaaaacgag ctcgaattca tcgatgatat cagatccact
     1621 agtggcctat gcggccgcca ccgcggtgga gctccaattc gccctatagt gagtcgtatt
     1681 acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     1741 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     1801 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg
     1861 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1921 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1981 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2041 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2101 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2161 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2221 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2281 aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     2341 tcacaccgca tagggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag
     2401 tatacatgca tgcgaattgc atatctttcc atagattttc aatttttgat gtctccatgg
     2461 tggtacagag aacttgtaaa caattcggtc cctacatgtg aggaaattcg ctgtgacact
     2521 tttatcactg aactccaaat ttaaaaaata gcataaaatt cgttatacag caaatctatg
     2581 tgttgcaatt aagaactaaa agatatagag tgcatatttt caagaaggat agtaagctgg
     2641 caaatcaagt cttagccttg ttcaatgggt tcttaccgta cttcttttcc aacttagaga
     2701 tgtaggtaat caatcttctg tgggacttag aatcaacgga caagatgaaa gacaaccaac
     2761 ctctaacatg ttcgatttcg gaggatttac cgcagaaaat atggtggatc ttggctctga
     2821 tttctttgta cttttctcta ccaataccaa ccttttcttg ggagataacc aaaccggtaa
     2881 cttttctttg ggatcttgga ccagaaatac aggtcttttt ggagttgatg accaaacctt
     2941 ctgatggaat aatggagaac aagaagtctc tggccttaac aactttcttc atagattgag
     3001 cagacaaggt caagtcatca gcgtatctag tgtatatcaa acctctagaa ccagcataac
     3061 cttgaattct gtagtccaac ttggagcaaa tcaagttagc caattttgga gaagatggag
     3121 caccttgtgg caacaagttc ttgtagcagc aaatcttggt caaaacagag gagatcaatc
     3181 tgttgtaacc caaagaatgg aaaacaccga aaaccttgtt agcagtcaaa gatgggaaga
     3241 aatcttccaa gtcgatgttc aaaatgaagt tggcaccaat atgtggagtg gcattgttca
     3301 agatggattg gtgcttttca aaaccgatag aaaatgggga ggaggataac ttgtccaaga
     3361 tgtttctcaa aacccaacct tgtaaagcct tcaattctct ggatggttgg tagatagttc
     3421 tcattctctt ttctggaccc ttcttttcaa cagtgtagat tctgtatctg aaatcggcgg
     3481 tgtagatcaa taatctcaag gtttcaacag aaattctggt agccttagac atatcgtgca
     3541 agttgttcat gactggcaaa cccaaatttc tcaatctgaa ggtgttcaag tattcagcgg
     3601 acttgacctt tcttttcttc tttggtggca tttatattga attttcaaaa attcttactt
     3661 tttttttgga tggacgcaaa gaagtttaat aatcatatta catggcatta ccaccatata
     3721 catatccata tctaatctta cttatatgtt gtggaaatgt aaagagcccc attatcttag
     3781 cctaaaaaaa ccttctcttt ggaactttca gtaatacgct taactgctca ttgctatatt
     3841 gaagtacgga ttagaagccg ccgagcgggc gacagccctc cgacggaaga ctctcctccg
     3901 tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctcgc gccgcactgc
     3961 tccgaacaat aaagattcta caatactagc ttttatggtt atgaagagga aaaattggca
     4021 gtaacctggc cccacaaacc ttcaaattaa cgaatcaaat taacaaccat aggatgataa
     4081 tgcgattagt tttttagcct tatttctggg gtaattaatc agcgaagcga tgatttttga
     4141 tctattaaca gatatataaa tggaaaagct gcataaccac tttaactaat actttcaaca
     4201 ttttcagttt gtattacttc ttattcaaat gtcataaaag tatcaacaaa aaattgttaa
     4261 tatacctcta tactttaacg tcaaggagaa aaaacaacta gtggatcccc cgggaaaaaa
     4321 atggacaaga agtactccat tgggctcgat atcggcacaa acagcgtcgg ttgggccgtc
     4381 attacggacg agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa taccgatcgc
     4441 cacagcataa agaagaacct cattggcgcc ctcctgttcg actccgggga gacggccgaa
     4501 gccacgcggc tcaaaagaac agcacggcgc agatataccc gcagaaagaa tcggatctgc
     4561 tacctgcagg agatctttag taatgagatg gctaaggtgg atgactcttt cttccatagg
     4621 ctggaggagt cctttttggt ggaggaggat aaaaagcacg agcgccaccc aatctttggc
     4681 aatatcgtgg acgaggtggc gtaccatgaa aagtacccaa ccatatatca tctgaggaag
     4741 aagcttgtag acagtactga taaggctgac ttgcggttga tctatctcgc gctggcgcat
     4801 atgatcaaat ttcggggaca cttcctcatc gagggggacc tgaacccaga caacagcgat
     4861 gtcgacaaac tctttatcca actggttcag acttacaatc agcttttcga agagaacccg
     4921 atcaacgcat ccggagttga cgccaaagca atcctgagcg ctaggctgtc caaatcccgg
     4981 cggctcgaaa acctcatcgc acagctccct ggggagaaga agaacggcct gtttggtaat
     5041 cttatcgccc tgtcactcgg gctgaccccc aactttaaat ctaacttcga cctggccgaa
     5101 gatgccaagc ttcaactgag caaagacacc tacgatgatg atctcgacaa tctgctggcc
     5161 cagatcggcg accagtacgc agaccttttt ttggcggcaa agaacctgtc agacgccatt
     5221 ctgctgagtg atattctgcg agtgaacacg gagatcacca aagctccgct gagcgctagt
     5281 atgatcaagc gctatgatga gcaccaccaa gacttgactt tgctgaaggc ccttgtcaga
     5341 cagcaactgc ctgagaagta caaggaaatt ttcttcgatc agtctaaaaa tggctacgcc
     5401 ggatacattg acggcggagc aagccaggag gaattttaca aatttattaa gcccatcttg
     5461 gaaaaaatgg acggcaccga ggagctgctg gtaaagctta acagagaaga tctgttgcgc
     5521 aaacagcgca ctttcgacaa tggaagcatc ccccaccaga ttcacctggg cgaactgcac
     5581 gctatcctca ggcggcaaga ggatttctac ccctttttga aagataacag ggaaaagatt
     5641 gagaaaatcc tcacatttcg gataccctac tatgtaggcc ccctcgcccg gggaaattcc
     5701 agattcgcgt ggatgactcg caaatcagaa gagaccatca ctccctggaa cttcgaggaa
     5761 gtcgtggata agggggcctc tgcccagtcc ttcatcgaaa ggatgactaa ctttgataaa
     5821 aatctgccta acgaaaaggt gcttcctaaa cactctctgc tgtacgagta cttcacagtt
     5881 tataacgagc tcaccaaggt caaatacgtc acagaaggga tgagaaagcc agcattcctg
     5941 tctggagagc agaagaaagc tatcgtggac ctcctcttca agacgaaccg gaaagttacc
     6001 gtgaaacagc tcaaagaaga ctatttcaaa aagattgaat gtttcgactc tgttgaaatc
     6061 agcggagtgg aggatcgctt caacgcatcc ctgggaacgt atcacgatct cctgaaaatc
     6121 attaaagaca aggacttcct ggacaatgag gagaacgagg acattcttga ggacattgtc
     6181 ctcaccctta cgttgtttga agatagggag atgattgaag aacgcttgaa aacttacgct
     6241 catctcttcg acgacaaagt catgaaacag ctcaagaggc gccgatatac aggatggggg
     6301 cggctgtcaa gaaaactgat caatgggatc cgagacaagc agagtggaaa gacaatcctg
     6361 gattttctta agtccgatgg atttgccaac cggaacttca tgcagttgat ccatgatgac
     6421 tctctcacct ttaaggagga catccagaaa gcacaagttt ctggccaggg ggacagtctt
     6481 cacgagcaca tcgctaatct tgcaggtagc ccagctatca aaaagggaat actgcagacc
     6541 gttaaggtcg tggatgaact cgtcaaagta atgggaaggc ataagcccga gaatatcgtt
     6601 atcgagatgg cccgagagaa ccaaactacc cagaagggac agaagaacag tagggaaagg
     6661 atgaagagga ttgaagaggg tataaaagaa ctggggtccc aaatccttaa ggaacaccca
     6721 gttgaaaaca cccagcttca gaatgagaag ctctacctgt actacctgca gaacggcagg
     6781 gacatgtacg tggatcagga actggacatc aatcggctct ccgactacga cgtggatcat
     6841 atcgtgcccc agtcttttct caaagatgat tctattgata ataaagtgtt gacaagatcc
     6901 gataaaaata gagggaagag tgataacgtc ccctcagaag aagttgtcaa gaaaatgaaa
     6961 aattattggc ggcagctgct gaacgccaaa ctgatcacac aacggaagtt cgataatctg
     7021 actaaggctg aacgaggtgg cctgtctgag ttggataaag ccggcttcat caaaaggcag
     7081 cttgttgaga cacgccagat caccaagcac gtggcccaaa ttctcgattc acgcatgaac
     7141 accaagtacg atgaaaatga caaactgatt cgagaggtga aagttattac tctgaagtct
     7201 aagctggtct cagatttcag aaaggacttt cagttttata aggtgagaga gatcaacaat
     7261 taccaccatg cgcatgatgc ctacctgaat gcagtggtag gcactgcact tatcaaaaaa
     7321 tatcccaagc ttgaatctga atttgtttac ggagactata aagtgtacga tgttaggaaa
     7381 atgatcgcaa agtctgagca ggaaataggc aaggccaccg ctaagtactt cttttacagc
     7441 aatattatga attttttcaa gaccgagatt acactggcca atggagagat tcggaagcga
     7501 ccacttatcg aaacaaacgg agaaacagga gaaatcgtgt gggacaaggg tagggatttc
     7561 gcgacagtcc ggaaggtcct gtccatgccg caggtgaaca tcgttaaaaa gaccgaagta
     7621 cagaccggag gcttctccaa ggaaagtatc ctcccgaaaa ggaacagcga caagctgatc
     7681 gcacgcaaaa aagattggga ccccaagaaa tacggcggat tcgattctcc tacagtcgct
     7741 tacagtgtac tggttgtggc caaagtggag aaagggaagt ctaaaaaact caaaagcgtc
     7801 aaggaactgc tgggcatcac aatcatggag cgatcaagct tcgaaaaaaa ccccatcgac
     7861 tttctcgagg cgaaaggata taaagaggtc aaaaaagacc tcatcattaa gcttcccaag
     7921 tactctctct ttgagcttga aaacggccgg aaacgaatgc tcgctagtgc gggcgagctg
     7981 cagaaaggta acgagctggc actgccctct aaatacgtta atttcttgta tctggccagc
     8041 cactatgaaa agctcaaagg gtctcccgaa gataatgagc agaagcagct gttcgtggaa
     8101 caacacaaac actaccttga tgagatcatc gagcaaataa gcgaattctc caaaagagtg
     8161 atcctcgccg acgctaacct cgataaggtg ctttctgctt acaataagca cagggataag
     8221 cccatcaggg agcaggcaga aaacattatc cacttgttta ctctgaccaa cttgggcgcg
     8281 cctgcagcct tcaagtactt cgacaccacc atagacagaa agcggtacac ctctacaaag
     8341 gaggtcctgg acgccacact gattcatcag tcaattacgg ggctctatga aacaagaatc
     8401 gacctctctc agctcggtgg agacagcagg gctgacccca agaagaagag gaaggtgtga
     8461 tctcttctcg agtcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat
     8521 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     8581 ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
     8641 acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac
     8701 gctcgaaggc tttaatttca tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     8761 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     8821 taataatggt ttcttagacg gatcgcttgc ctgtaactta cacgcgcctc gtatctttta
     8881 atgatggaat aatttgggaa tttactctgt gtttatttat ttttatgttt tgtatttgga
     8941 ttttagaaag taaataaaga aggtagaaga gttacggaat gaagaaaaaa aaataaacaa
     9001 aggtttaaaa aatttcaaca aaaagcgtac tttacatata tatttattag acaagaaaag
     9061 cagattaaat agatatacat tcgattaacg ataagtaaaa tgtaaaatca caggattttc
     9121 gtgtgtggtc ttctacacag acaagatgaa acaattcggc attaatacct gagagcagga
     9181 agagcaagat aaaaggtagt atttgttggc gatcccccta gagtctttta catcttcgga
     9241 aaacaaaaac tattttttct ttaatttctt tttttacttt ctatttttaa tttatatatt
     9301 tatattaaaa aatttaaatt ataattattt ttatagcacg tgatgaaaag gacccaggtg
     9361 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     9421 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     9481 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     9541 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     9601 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     9661 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     9721 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     9781 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     9841 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     9901 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     9961 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
    10021 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
    10081 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
    10141 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
    10201 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
    10261 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
    10321 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
    10381 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
    10441 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
    10501 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
    10561 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
    10621 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
    10681 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
    10741 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
    10801 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
    10861 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
    10921 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
    10981 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
    11041 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
    11101 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
    11161 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
    11221 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
    11281 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
    11341 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
    11401 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
    11461 gtggaattgt gagcggataa caat
//
