LOCUS       sequence-403782-       10025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TEF1p-Cas9-CYC1t and SNR52p-Not1-SUP4t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10025)
  AUTHORS   Zhao Y, Coelho C, Lauer S, Majewski M, Laurent JM, Brosh R, Boeke JD
  TITLE     CREEPY: CRISPR-mediated editing of synthetic episomes in yeast.
  JOURNAL   Nucleic Acids Res. 2023 Jun 16:gkad491. doi: 10.1093/nar/gkad491.
  PUBMED    37326023
REFERENCE   2  (bases 1 to 10025)
  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: "Nucleic 
            Acids Res. 2023 Jun 16:gkad491. doi: 10.1093/nar/gkad491."
COMMENT     Sequence Label: pYZ462
FEATURES             Location/Qualifiers
     source          1..10025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..24)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    65..568
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        605..709
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(710..778,779..1570)
                     /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(928..947)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1741..2329
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2230..2249
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2483..2500
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2617..2638
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2653..2658,2659..2676,2677..2683)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2691..2707
                     /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     2696..2718
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2715..2731
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2715..2731
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2750..2770
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2752..2770
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2788..3186
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     3190..3206
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3190..3206
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3218..7321
                     /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             3218..7321
                     /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             3218..7321
                     /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             7334..7354
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7370..7617
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7386..7404)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        7632..7900
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        7909..7983
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      7988..8007
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     primer_bind     complement(8056..8072)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8057..8073)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(8097..8116)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8098..8116)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8126..8143)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8126..8142)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8135..8157)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8283..8738
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8370..8389)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8580..8601
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(8869..9672)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(9673..9888)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(9912..9931)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
       61 gacggatcgc ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg gaataatttg
      121 ggaatttact ctgtgtttat ttatttttat gttttgtatt tggattttag aaagtaaata
      181 aagaaggtag aagagttacg gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc
      241 aacaaaaagc gtactttaca tatatattta ttagacaaga aaagcagatt aaatagatat
      301 acattcgatt aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac
      361 acagacaaga tgaaacaatt cggcattaat acctgagagc aggaagagca agataaaagg
      421 tagtatttgt tggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt
      481 ttctttaatt tcttttttta ctttctattt ttaatttata tatttatatt aaaaaattta
      541 aattataatt atttttatag cacgtgatga aaaggaccca ggtggcactt ttcggggaaa
      601 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      661 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
      721 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
      781 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
      841 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
      901 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
      961 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1021 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1081 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1141 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1201 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1261 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1321 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1381 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggatctc gcggtatcat
     1441 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1501 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     1561 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     1621 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1681 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1741 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1801 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1861 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1921 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1981 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2041 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2101 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2161 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2221 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2281 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2341 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2401 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2461 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     2521 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     2581 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     2641 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     2701 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     2761 tcactaaagg gaacaaaagc tggagctcat agcttcaaaa tgtttctact ccttttttac
     2821 tcttccagat tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag
     2881 catactaaat ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt
     2941 tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat
     3001 ttttatcacg tttctttttc ttgaaaattt tttttttgat ttttttctct ttcgatgacc
     3061 tcccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag tttcattttt
     3121 cttgttctat tacaactttt tttacttctt gctcattaga aagaaagcat agcaatctaa
     3181 tctaagtttt ctagaactag tggatccccc gggaaaaatg gacaagaagt actccattgg
     3241 gctcgatatc ggcacaaaca gcgtcggttg ggccgtcatt acggacgagt acaaggtgcc
     3301 gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac agcataaaga agaacctcat
     3361 tggcgccctc ctgttcgact ccggggagac ggccgaagcc acgcggctca aaagaacagc
     3421 acggcgcaga tatacccgca gaaagaatcg gatctgctac ctgcaggaga tctttagtaa
     3481 tgagatggct aaggtggatg actctttctt ccataggctg gaggagtcct ttttggtgga
     3541 ggaggataaa aagcacgagc gccacccaat ctttggcaat atcgtggacg aggtggcgta
     3601 ccatgaaaag tacccaacca tatatcatct gaggaagaag cttgtagaca gtactgataa
     3661 ggctgacttg cggttgatct atctcgcgct ggcgcatatg atcaaatttc ggggacactt
     3721 cctcatcgag ggggacctga acccagacaa cagcgatgtc gacaaactct ttatccaact
     3781 ggttcagact tacaatcagc ttttcgaaga gaacccgatc aacgcatccg gagttgacgc
     3841 caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca
     3901 gctccctggg gagaagaaga acggcctgtt tggtaatctt atcgccctgt cactcgggct
     3961 gacccccaac tttaaatcta acttcgacct ggccgaagat gccaagcttc aactgagcaa
     4021 agacacctac gatgatgatc tcgacaatct gctggcccag atcggcgacc agtacgcaga
     4081 cctttttttg gcggcaaaga acctgtcaga cgccattctg ctgagtgata ttctgcgagt
     4141 gaacacggag atcaccaaag ctccgctgag cgctagtatg atcaagcgct atgatgagca
     4201 ccaccaagac ttgactttgc tgaaggccct tgtcagacag caactgcctg agaagtacaa
     4261 ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga tacattgacg gcggagcaag
     4321 ccaggaggaa ttttacaaat ttattaagcc catcttggaa aaaatggacg gcaccgagga
     4381 gctgctggta aagcttaaca gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg
     4441 aagcatcccc caccagattc acctgggcga actgcacgct atcctcaggc ggcaagagga
     4501 tttctacccc tttttgaaag ataacaggga aaagattgag aaaatcctca catttcggat
     4561 accctactat gtaggccccc tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa
     4621 atcagaagag accatcactc cctggaactt cgaggaagtc gtggataagg gggcctctgc
     4681 ccagtccttc atcgaaagga tgactaactt tgataaaaat ctgcctaacg aaaaggtgct
     4741 tcctaaacac tctctgctgt acgagtactt cacagtttat aacgagctca ccaaggtcaa
     4801 atacgtcaca gaagggatga gaaagccagc attcctgtct ggagagcaga agaaagctat
     4861 cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg aaacagctca aagaagacta
     4921 tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa
     4981 cgcatccctg ggaacgtatc acgatctcct gaaaatcatt aaagacaagg acttcctgga
     5041 caatgaggag aacgaggaca ttcttgagga cattgtcctc acccttacgt tgtttgaaga
     5101 tagggagatg attgaagaac gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat
     5161 gaaacagctc aagaggcgcc gatatacagg atgggggcgg ctgtcaagaa aactgatcaa
     5221 tgggatccga gacaagcaga gtggaaagac aatcctggat tttcttaagt ccgatggatt
     5281 tgccaaccgg aacttcatgc agttgatcca tgatgactct ctcaccttta aggaggacat
     5341 ccagaaagca caagtttctg gccaggggga cagtcttcac gagcacatcg ctaatcttgc
     5401 aggtagccca gctatcaaaa agggaatact gcagaccgtt aaggtcgtgg atgaactcgt
     5461 caaagtaatg ggaaggcata agcccgagaa tatcgttatc gagatggccc gagagaacca
     5521 aactacccag aagggacaga agaacagtag ggaaaggatg aagaggattg aagagggtat
     5581 aaaagaactg gggtcccaaa tccttaagga acacccagtt gaaaacaccc agcttcagaa
     5641 tgagaagctc tacctgtact acctgcagaa cggcagggac atgtacgtgg atcaggaact
     5701 ggacatcaat cggctctccg actacgacgt ggatcatatc gtgccccagt cttttctcaa
     5761 agatgattct attgataata aagtgttgac aagatccgat aaaaatagag ggaagagtga
     5821 taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa
     5881 cgccaaactg atcacacaac ggaagttcga taatctgact aaggctgaac gaggtggcct
     5941 gtctgagttg gataaagccg gcttcatcaa aaggcagctt gttgagacac gccagatcac
     6001 caagcacgtg gcccaaattc tcgattcacg catgaacacc aagtacgatg aaaatgacaa
     6061 actgattcga gaggtgaaag ttattactct gaagtctaag ctggtctcag atttcagaaa
     6121 ggactttcag ttttataagg tgagagagat caacaattac caccatgcgc atgatgccta
     6181 cctgaatgca gtggtaggca ctgcacttat caaaaaatat cccaagcttg aatctgaatt
     6241 tgtttacgga gactataaag tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga
     6301 aataggcaag gccaccgcta agtacttctt ttacagcaat attatgaatt ttttcaagac
     6361 cgagattaca ctggccaatg gagagattcg gaagcgacca cttatcgaaa caaacggaga
     6421 aacaggagaa atcgtgtggg acaagggtag ggatttcgcg acagtccgga aggtcctgtc
     6481 catgccgcag gtgaacatcg ttaaaaagac cgaagtacag accggaggct tctccaagga
     6541 aagtatcctc ccgaaaagga acagcgacaa gctgatcgca cgcaaaaaag attgggaccc
     6601 caagaaatac ggcggattcg attctcctac agtcgcttac agtgtactgg ttgtggccaa
     6661 agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat
     6721 catggagcga tcaagcttcg aaaaaaaccc catcgacttt ctcgaggcga aaggatataa
     6781 agaggtcaaa aaagacctca tcattaagct tcccaagtac tctctctttg agcttgaaaa
     6841 cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag aaaggtaacg agctggcact
     6901 gccctctaaa tacgttaatt tcttgtatct ggccagccac tatgaaaagc tcaaagggtc
     6961 tcccgaagat aatgagcaga agcagctgtt cgtggaacaa cacaaacact accttgatga
     7021 gatcatcgag caaataagcg aattctccaa aagagtgatc ctcgccgacg ctaacctcga
     7081 taaggtgctt tctgcttaca ataagcacag ggataagccc atcagggagc aggcagaaaa
     7141 cattatccac ttgtttactc tgaccaactt gggcgcgcct gcagccttca agtacttcga
     7201 caccaccata gacagaaagc ggtacacctc tacaaaggag gtcctggacg ccacactgat
     7261 tcatcagtca attacggggc tctatgaaac aagaatcgac ctctctcagc tcggtggaga
     7321 cagcagggct gaccccaaga agaagaggaa ggtgtgatct cttctcgagt catgtaatta
     7381 gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga
     7441 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
     7501 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
     7561 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcgca
     7621 tgcctggagc ttctttgaaa agataatgta tgattatgct ttcactcata tttatacaga
     7681 aacttgatgt tttctttcga gtatatacaa ggtgattaca tgtacgtttg aagtacaact
     7741 ctagattttg tagtgccctc ttgggctagc ggtaaaggtg cgcatttttt cacaccctac
     7801 aatgttctgt tcaaaagatt ttggtcaaac gctgtagaag tgaaagttgg tgcgcatgtt
     7861 tcggcgttcg aaacttctcc gcagtgaaag ataaatgatc gcggccgcgt tttagagcta
     7921 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     7981 gtggtgcttt ttttgttttt tatgtcttcg agtcatgtaa ttagttatgt cacgcgaatt
     8041 cgatatcaag cttatcgata ccgtcgacct cgaggggggg cccggtaccc aattcgccct
     8101 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     8161 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     8221 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     8281 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     8341 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     8401 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     8461 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     8521 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     8581 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     8641 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     8701 taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
     8761 acgcatctgt gcggtatttc acaccgcata gggtaataac tgatataatt aaattgaagc
     8821 tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg
     8881 gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt
     8941 agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga
     9001 caccacatca tccacggttc tatactgttg acccaatgca tctcccttgt catctaaacc
     9061 cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg
     9121 agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt
     9181 atattctcca gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct
     9241 aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac
     9301 cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta
     9361 ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt
     9421 gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa
     9481 gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag
     9541 taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat
     9601 gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt
     9661 agctttcgac atgatttatc ttcgtttcct gcaggttttt gttctgtgca gttgggttaa
     9721 gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag
     9781 tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca
     9841 aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa ggtggtatgg
     9901 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     9961 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
    10021 gtgac
//
