LOCUS       sequence-459850-       13557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast CEN plasmid for estradiol-inducible expression of spCas9 and 
            Ec86 reverse transcriptase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13557)
  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 13557)
  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-Z3-Kan
FEATURES             Location/Qualifiers
     source          1..13557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             54..74
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     rep_origin      complement(3406..3861)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3543..3564)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3755..3774
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3987..4009
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4001..4018
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4002..4018
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4025..4044
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4025..4043
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            complement(4160..5516)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(4160..4357)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(4363..5172)
                     /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     5086..5105
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5173..5516)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        complement(5644..5662)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5683..5699)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5683..5699)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5696..5718)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5707..5723
                     /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(5731..5737,5738..5755,5756..5761))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5776..5797
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5914..5931)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6085..6673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6165..6184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6844..7635,7636..7704))
                     /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     7467..7486
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7705..7809)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7846..8349
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     8390..8408
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8446..8468)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     terminator      complement(8482..8671)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     8683..8701
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(8733..8753)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8766..12869)
                     /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             complement(8766..12869)
                     /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             complement(8766..12869)
                     /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"
     primer_bind     complement(12912..12935)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
        1 aaaaaaaaag taagaatttt tgaaaattca atataaattc aatataaatg ccaccaaaga
       61 agaaaagaaa ggtcaagtcc gctgaatact tgaacacctt cagattgaga aatttgggtt
      121 tgccagtcat gaacaacttg cacgatatgt ctaaggctac cagaatttct gttgaaacct
      181 tgagattatt gatctacacc gccgatttca gatacagaat ctacactgtt gaaaagaagg
      241 gtccagaaaa gagaatgaga actatctacc aaccatccag agaattgaag gctttacaag
      301 gttgggtttt gagaaacatc ttggacaagt tatcctcctc cccattttct atcggttttg
      361 aaaagcacca atccatcttg aacaatgcca ctccacatat tggtgccaac ttcattttga
      421 acatcgactt ggaagatttc ttcccatctt tgactgctaa caaggttttc ggtgttttcc
      481 attctttggg ttacaacaga ttgatctcct ctgttttgac caagatttgc tgctacaaga
      541 acttgttgcc acaaggtgct ccatcttctc caaaattggc taacttgatt tgctccaagt
      601 tggactacag aattcaaggt tatgctggtt ctagaggttt gatatacact agatacgctg
      661 atgacttgac cttgtctgct caatctatga agaaagttgt taaggccaga gacttcttgt
      721 tctccattat tccatcagaa ggtttggtca tcaactccaa aaagacctgt atttctggtc
      781 caagatccca aagaaaagtt accggtttgg ttatctccca agaaaaggtt ggtattggta
      841 gagaaaagta caaagaaatc agagccaaga tccaccatat tttctgcggt aaatcctccg
      901 aaatcgaaca tgttagaggt tggttgtctt tcatcttgtc cgttgattct aagtcccaca
      961 gaagattgat tacctacatc tctaagttgg aaaagaagta cggtaagaac ccattgaaca
     1021 aggctaagac ttgatttgcc agcttactat ccttcttgaa aatatgcact ctatatcttt
     1081 tagttcttaa ttgcaacaca tagatttgct gtataacgaa ttttatgcta ttttttaaat
     1141 ttggagttca gtgataaaag tgtcacagcg aatttcctca catgtaggga ccgaattgtt
     1201 tacaagttct ctgtaccacc atggagacat caaaaattga aaatctatgg aaagatatgc
     1261 aattcgcatg catcccttta aaaacatatg cctcacccta acatattttc caattaaccc
     1321 tcaatatttc tctgtcaccc ggcctctatt ttccattttc ttctttaccc gccacgcgtt
     1381 tttttctttc aaattttttt cttccttctt ctttttcttc cacgtcctct tgcataaata
     1441 aataaaccgt tttgaaacca aactcgcctc tctctctcct ttttgaaata tttttgggtt
     1501 tgtttgatcc tttccttccc aatctctctt gtttaatata tattcattta tatcacgctc
     1561 tctttttatc ttcctttttt tcctctctct tgtattcttc cttccccttt ctactcaaac
     1621 caagaagaaa aagaaaaggt caatctttgt taaagaatag gatcttctac tacatcagct
     1681 tttagatttt tcacgcttac tgcttttttc ttcccaagat cgaaaattta ctgaattaac
     1741 aatgggaacg agaccctacg cttgccctgt agaaagttgt gataggagat tcagcagatc
     1801 tgacgagctg actaggcata ttcgtattca caccggccaa aagcctttcc aatgtagaat
     1861 ctgtatgaga aacttcagca gaagtgacca tttgactacg cacattagaa cccacaccgg
     1921 cgagaagccg ttcgcgtgtg acatttgtgg aagaaagttc gccagatctg atgaaagaaa
     1981 gagacataca aagatccata caggaggtgg gggcactccg gcagcagcct caaccctaga
     2041 agatccaagc gctggagaca tgagggcagc caacttgtgg ccaagcccat taatgatcaa
     2101 aagatctaag aagaattctc ttgcactgtc tttgaccgct gatcaaatgg tttccgcact
     2161 acttgatgct gaaccaccta tattgtacag tgaatacgac ccaactcgtc ctttttccga
     2221 ggcttccatg atgggtcttt tgacaaatct agcagacagg gaattagttc atatgattaa
     2281 ttgggccaaa agagttcccg gctttgttga cttgacatta catgaccagg tacatcttct
     2341 tgaatgcgcg tggttagaaa tattgatgat cggattagtg tggagaagta tggagcaccc
     2401 tgtaaaatta ttatttgcac ctaatttatt attggacagg aaccaaggta aatgcgtaga
     2461 aggtatggtt gagatttttg atatgcttct tgccactagc tccaggttca gaatgatgaa
     2521 tttgcaagga gaagaattcg tttgcctgaa atcaattatt ttgttaaata gcggtgtcta
     2581 tacattttta agctcaactc tgaaaagcct ggaagagaaa gaccatatac atagagtgtt
     2641 ggacaagatc actgatacat tgattcacct tatggcgaaa gcaggattga ctctgcagca
     2701 acagcatcag aggctagctc agttactact aattttgtca cacattagac atatgtctaa
     2761 caagggtatg gagcacctgt attctatgaa gtgtaaaaac gtcgttcctc tgtatgattt
     2821 attactagaa atgctggacg ctcacaggtt gcatgcacca acttctagag gcggagcctc
     2881 cgttgaagaa accgaccaat cccacctagc gacagcaggg tctacatcta gcgaactgca
     2941 tttggacggt gaagacgttg ccatggccca cgctgacgct ttagacgatt ttgatttaga
     3001 tatgttgggt gacggggact ctccagggcc tggcttcact ccacatgatt cagcacctta
     3061 cggtgcgctg gatatggctg acttcgaatt tgagcagatg ttcaccgatg ctctgggtat
     3121 tgatgagtat ggtggttagt agtaagctgg ttcttctttc tgccatttaa ttgtagactt
     3181 gttttctatt taatatcact gtatcctgta gcgtttgtat acattccgta atagaaggtc
     3241 cgatacttta ctttcatatc agtcgcagtt tgccacttga ggggagatcg acgcatgcat
     3301 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta ccctatgcgg
     3361 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta
     3421 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3481 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3541 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3601 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3661 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3721 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3781 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3841 atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg
     3901 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     3961 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     4021 attgtaatac gactcactat agggcgaatt ggagctccac cgcggtggcg gccgcatagg
     4081 ccactagtgg atctgatatc atcgatgaat tcgagctcgt tttcgacact ggatggcggc
     4141 gttagtatcg aatcgacagc agtatagcga ccagcattca catacgattg acgcatgata
     4201 ttactttctg cgcacttaac ttcgcatctg ggcagatgat gtcgaggcga aaaaaaatat
     4261 aaatcacgct aacatttgat taaaatagaa caactacaat ataaaaaaac tatacaaatg
     4321 acaagttctt gaaaacaaga atctttttat tgtcagtact gattagaaaa actcatcgag
     4381 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     4441 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     4501 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     4561 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     4621 caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     4681 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     4741 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     4801 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     4861 tgctgttttg ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     4921 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     4981 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     5041 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     5101 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctcgaaa cgtgagtctt
     5161 ttccttaccc atggttgttt atgttcggat gtgatgtgag aactgtatcc tagcaagatt
     5221 ttaaaaggaa gtatatgaaa gaagaacctc agtggcaaat cctaaccttt tatatttctc
     5281 tacaggggcg cggcgtgggg acaattcaac gcgtctgtga ggggagcgtt tccctgctcg
     5341 caggtccgca gcgaggagcc gtaatttttg cttcgcgccg tgcggccatc aaaatgtatg
     5401 gatgcaaatg attatacatg gggatgtatg ggctaaatgt acgggcgaca gtcacatcat
     5461 gcccctgagc tgcgcacgtc aagactgtca aggagggtat tctgggcctc catgtcgctg
     5521 gccgggtgac ccggcgggga cgaggcaagc taaacagatc tggcgcgcct taattaaccc
     5581 ggggatccgt cgacctgcag cgtacgaagc ttcagctggc ggccgctcta gcagcttttg
     5641 ttccctttag tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt
     5701 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5761 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5821 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5881 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5941 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6001 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6061 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6121 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6181 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6241 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6301 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6361 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6421 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6481 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6541 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6601 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6661 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6721 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6781 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6841 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6901 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6961 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7021 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7081 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7141 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7201 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7261 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7321 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7381 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7441 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7501 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7561 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7621 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7681 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7741 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7801 ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa
     7861 aataattata atttaaattt tttaatataa atatataaat taaaaataga aagtaaaaaa
     7921 agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct agggggatcg
     7981 ccaacaaata ctacctttta tcttgctctt cctgctctca ggtattaatg ccgaattgtt
     8041 tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat tttacttatc
     8101 gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat atatgtaaag
     8161 tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca ttccgtaact
     8221 cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa taaataaaca
     8281 cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt aagttacagg
     8341 caagcgatcc gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     8401 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     8461 gctcccggat gaaattaaag ccttcgagcg tcccaaaacc ttctcaagca aggttttcag
     8521 tataatgtta catgcgtaca cgcgtctgta cagaaaaaaa agaaaaattt gaaatataaa
     8581 taacgttctt aatactaaca taactataaa aaaataaata gggacctaga cttcaggttg
     8641 tctaactcct tccttttcgg ttagagcgga tgtgggggga gggcgtgaat gtaagcgtga
     8701 cataactaat tacatgactc gagaagagat cacaccttcc tcttcttctt ggggtcagcc
     8761 ctgctgtctc caccgagctg agagaggtcg attcttgttt catagagccc cgtaattgac
     8821 tgatgaatca gtgtggcgtc caggacctcc tttgtagagg tgtaccgctt tctgtctatg
     8881 gtggtgtcga agtacttgaa ggctgcaggc gcgcccaagt tggtcagagt aaacaagtgg
     8941 ataatgtttt ctgcctgctc cctgatgggc ttatccctgt gcttattgta agcagaaagc
     9001 accttatcga ggttagcgtc ggcgaggatc actcttttgg agaattcgct tatttgctcg
     9061 atgatctcat caaggtagtg tttgtgttgt tccacgaaca gctgcttctg ctcattatct
     9121 tcgggagacc ctttgagctt ttcatagtgg ctggccagat acaagaaatt aacgtattta
     9181 gagggcagtg ccagctcgtt acctttctgc agctcgcccg cactagcgag cattcgtttc
     9241 cggccgtttt caagctcaaa gagagagtac ttgggaagct taatgatgag gtcttttttg
     9301 acctctttat atcctttcgc ctcgagaaag tcgatggggt ttttttcgaa gcttgatcgc
     9361 tccatgattg tgatgcccag cagttccttg acgcttttga gttttttaga cttccctttc
     9421 tccactttgg ccacaaccag tacactgtaa gcgactgtag gagaatcgaa tccgccgtat
     9481 ttcttggggt cccaatcttt tttgcgtgcg atcagcttgt cgctgttcct tttcgggagg
     9541 atactttcct tggagaagcc tccggtctgt acttcggtct ttttaacgat gttcacctgc
     9601 ggcatggaca ggaccttccg gactgtcgcg aaatccctac ccttgtccca cacgatttct
     9661 cctgtttctc cgtttgtttc gataagtggt cgcttccgaa tctctccatt ggccagtgta
     9721 atctcggtct tgaaaaaatt cataatattg ctgtaaaaga agtacttagc ggtggccttg
     9781 cctatttcct gctcagactt tgcgatcatt ttcctaacat cgtacacttt atagtctccg
     9841 taaacaaatt cagattcaag cttgggatat tttttgataa gtgcagtgcc taccactgca
     9901 ttcaggtagg catcatgcgc atggtggtaa ttgttgatct ctctcacctt ataaaactga
     9961 aagtcctttc tgaaatctga gaccagctta gacttcagag taataacttt cacctctcga
    10021 atcagtttgt cattttcatc gtacttggtg ttcatgcgtg aatcgagaat ttgggccacg
    10081 tgcttggtga tctggcgtgt ctcaacaagc tgccttttga tgaagccggc tttatccaac
    10141 tcagacaggc cacctcgttc agccttagtc agattatcga acttccgttg tgtgatcagt
    10201 ttggcgttca gcagctgccg ccaataattt ttcattttct tgacaacttc ttctgagggg
    10261 acgttatcac tcttccctct atttttatcg gatcttgtca acactttatt atcaatagaa
    10321 tcatctttga gaaaagactg gggcacgata tgatccacgt cgtagtcgga gagccgattg
    10381 atgtccagtt cctgatccac gtacatgtcc ctgccgttct gcaggtagta caggtagagc
    10441 ttctcattct gaagctgggt gttttcaact gggtgttcct taaggatttg ggaccccagt
    10501 tcttttatac cctcttcaat cctcttcatc ctttccctac tgttcttctg tcccttctgg
    10561 gtagtttggt tctctcgggc catctcgata acgatattct cgggcttatg ccttcccatt
    10621 actttgacga gttcatccac gaccttaacg gtctgcagta ttcccttttt gatagctggg
    10681 ctacctgcaa gattagcgat gtgctcgtga agactgtccc cctggccaga aacttgtgct
    10741 ttctggatgt cctccttaaa ggtgagagag tcatcatgga tcaactgcat gaagttccgg
    10801 ttggcaaatc catcggactt aagaaaatcc aggattgtct ttccactctg cttgtctcgg
    10861 atcccattga tcagttttct tgacagccgc ccccatcctg tatatcggcg cctcttgagc
    10921 tgtttcatga ctttgtcgtc gaagagatga gcgtaagttt tcaagcgttc ttcaatcatc
    10981 tccctatctt caaacaacgt aagggtgagg acaatgtcct caagaatgtc ctcgttctcc
    11041 tcattgtcca ggaagtcctt gtctttaatg attttcagga gatcgtgata cgttcccagg
    11101 gatgcgttga agcgatcctc cactccgctg atttcaacag agtcgaaaca ttcaatcttt
    11161 ttgaaatagt cttctttgag ctgtttcacg gtaactttcc ggttcgtctt gaagaggagg
    11221 tccacgatag ctttcttctg ctctccagac aggaatgctg gctttctcat cccttctgtg
    11281 acgtatttga ccttggtgag ctcgttataa actgtgaagt actcgtacag cagagagtgt
    11341 ttaggaagca ccttttcgtt aggcagattt ttatcaaagt tagtcatcct ttcgatgaag
    11401 gactgggcag aggccccctt atccacgact tcctcgaagt tccagggagt gatggtctct
    11461 tctgatttgc gagtcatcca cgcgaatctg gaatttcccc gggcgagggg gcctacatag
    11521 tagggtatcc gaaatgtgag gattttctca atcttttccc tgttatcttt caaaaagggg
    11581 tagaaatcct cttgccgcct gaggatagcg tgcagttcgc ccaggtgaat ctggtggggg
    11641 atgcttccat tgtcgaaagt gcgctgtttg cgcaacagat cttctctgtt aagctttacc
    11701 agcagctcct cggtgccgtc cattttttcc aagatgggct taataaattt gtaaaattcc
    11761 tcctggcttg ctccgccgtc aatgtatccg gcgtagccat ttttagactg atcgaagaaa
    11821 atttccttgt acttctcagg cagttgctgt ctgacaaggg ccttcagcaa agtcaagtct
    11881 tggtggtgct catcatagcg cttgatcata ctagcgctca gcggagcttt ggtgatctcc
    11941 gtgttcactc gcagaatatc actcagcaga atggcgtctg acaggttctt tgccgccaaa
    12001 aaaaggtctg cgtactggtc gccgatctgg gccagcagat tgtcgagatc atcatcgtag
    12061 gtgtctttgc tcagttgaag cttggcatct tcggccaggt cgaagttaga tttaaagttg
    12121 ggggtcagcc cgagtgacag ggcgataaga ttaccaaaca ggccgttctt cttctcccca
    12181 gggagctgtg cgatgaggtt ttcgagccgc cgggatttgg acagcctagc gctcaggatt
    12241 gctttggcgt caactccgga tgcgttgatc gggttctctt cgaaaagctg attgtaagtc
    12301 tgaaccagtt ggataaagag tttgtcgaca tcgctgttgt ctgggttcag gtccccctcg
    12361 atgaggaagt gtccccgaaa tttgatcata tgcgccagcg cgagatagat caaccgcaag
    12421 tcagccttat cagtactgtc tacaagcttc ttcctcagat gatatatggt tgggtacttt
    12481 tcatggtacg ccacctcgtc cacgatattg ccaaagattg ggtggcgctc gtgcttttta
    12541 tcctcctcca ccaaaaagga ctcctccagc ctatggaaga aagagtcatc caccttagcc
    12601 atctcattac taaagatctc ctgcaggtag cagatccgat tctttctgcg ggtatatctg
    12661 cgccgtgctg ttcttttgag ccgcgtggct tcggccgtct ccccggagtc gaacaggagg
    12721 gcgccaatga ggttcttctt tatgctgtgg cgatcggtat tgcccagaac tttgaatttt
    12781 ttgctcggca ccttgtactc gtccgtaatg acggcccaac cgacgctgtt tgtgccgata
    12841 tcgagcccaa tggagtactt cttgtccatt tttttcccgg gggatccact agtttatagt
    12901 tttttctcct tgacgttaaa gtatagaggt atattaacaa ttttttgttg atacttttat
    12961 gacatttgaa taagaagtaa tacaaactga aaatgttgaa agtattagtt aaagtggtta
    13021 tgcagctttt ccatttatat atctgttaat agatcaaaaa tcatcgcttc gctgattaat
    13081 taccccagaa ataaggctaa aaaactaatc gcattatcat cctatggttg ttaatttgat
    13141 tcgttaattt gaaggtttgt ggggccaggt tactgccaat ttttcctctt cataaccata
    13201 aaagctagta ttgtagaatc tttattgttc ggagcagtgc ggcgcgaggc acatctgcgt
    13261 ttcaggaacg cgaccggtga agacgaggac gcacggtcta gacgcccacg cacgcccacg
    13321 cccgcccacg cacgcccacg cccgcccacg cacgcccacg cggccgcact tcaatatagc
    13381 aatgagcagt taagcgtatt actgaaagtt ccaaagagaa ggttttttta ggctaagata
    13441 atggggctct ttacatttcc acaacatata agtaagatta gatatggata tgtatatgga
    13501 tatgtatatg gtggtaatgc catgtaatat gattattaaa cttctttgcg tccatcc
//
