LOCUS       sequence-409168-       12419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TEF1p-Cas9(noBsmBI)-CYC1t and tRNA-GFP-SUP4t with 2µ-URA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12419)
  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 12419)
  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: pYZ960
FEATURES             Location/Qualifiers
     source          1..12419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..24)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      65..1407
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    397..444
                     /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)."
     promoter        1434..1538
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1539..1607,1608..2399)
                     /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(1757..1776)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2570..3158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3059..3078
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3312..3329
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3446..3467
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3482..3487,3488..3505,3506..3512)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3520..3536
                     /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     3525..3547
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3544..3560
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3544..3560
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3579..3599
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3581..3599
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3617..4015
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     4019..4035
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4019..4035
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             4047..8150
                     /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             4047..8150
                     /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             4047..8150
                     /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             8163..8183
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8199..8446
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(8215..8233)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     RBS             9108..9119
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             9126..9839
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      9851..9922
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      9938..9965
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        9979..10054
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(10492..10511)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10493..10511)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10521..10538)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10521..10537)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10530..10552)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10678..11133
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10765..10784)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10975..10996
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(11264..12067)
                     /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(12068..12282)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(12306..12325)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
       61 gtatgatcca atatcaaagg aaatgatagc attgaaggat gagactaatc caattgagga
      121 gtggcagcat atagaacagc taaagggtag tgctgaagga agcatacgat accccgcatg
      181 gaatgggata atatcacagg aggtactaga ctacctttca tcctacataa atagacgcat
      241 ataagtacgc atttaagcat aaacacgcac tatgccgttc ttctcatgta tatatatata
      301 caggcaacac gcagatatag gtgcgacgtg aacagtgagc tgtatgtgcg cagctcgcgt
      361 tgcattttcg gaagcgctcg ttttcggaaa cgctttgaag ttcctattcc gaagttccta
      421 ttctctagaa agtataggaa cttcagagcg cttttgaaaa ccaaaagcgc tctgaagacg
      481 cactttcaaa aaaccaaaaa cgcaccggac tgtaacgagc tactaaaata ttgcgaatac
      541 cgcttccaca aacattgctc aaaagtatct ctttgctata tatctctgtg ctatatccct
      601 atataaccta cccatccacc tttcgctcct tgaacttgca tctaaactcg acctctacat
      661 tttttatgtt tatctctagt attactcttt agacaaaaaa attgtagtaa gaactattca
      721 tagagtgaat cgaaaacaat acgaaaatgt aaacatttcc tatacgtagt atatagagac
      781 aaaatagaag aaaccgttca taattttctg accaatgaag aatcatcaac gctatcactt
      841 tctgttcaca aagtatgcgc aatccacatc ggtatagaat ataatcgggg atgcctttat
      901 cttgaaaaaa tgcacccgca gcttcgctag taatcagtaa acgcgggaag tggagtcagg
      961 ctttttttat ggaagagaaa atagacacca aagtagcctt cttctaacct taacggacct
     1021 acagtgcaaa aagttatcaa gagactgcat tatagagcgc acaaaggaga aaaaaagtaa
     1081 tctaagatgc tttgttagaa aaatagcgct ctcgggatgc atttttgtag aacaaaaaag
     1141 aagtatagat tctttgttgg taaaatagcg ctctcgcgtt gcatttctgt tctgtaaaaa
     1201 tgcagctcag attctttgtt tgaaaaatta gcgctctcgc gttgcatttt tgttttacaa
     1261 aaatgaagca cagattcttc gttggtaaaa tagcgctttc gcgttgcatt tctgttctgt
     1321 aaaaatgcag ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc atttttgttc
     1381 tacaaaatga agcacagatg cttcgttcag gtggcacttt tcggggaaat gtgcgcggaa
     1441 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1501 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     1561 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1621 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     1681 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1741 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     1801 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     1861 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     1921 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     1981 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     2041 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     2101 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     2161 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     2221 ttattgctga taaatctgga gccggtgagc gtggatctcg cggtatcatt gcagcactgg
     2281 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     2341 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     2401 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     2461 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2521 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2581 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2641 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2701 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2761 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2821 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2881 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2941 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3001 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3061 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3121 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3181 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3241 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3301 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     3361 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     3421 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     3481 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     3541 acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg
     3601 aacaaaagct ggagctcata gcttcaaaat gtttctactc cttttttact cttccagatt
     3661 ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc caagcacagc atactaaatt
     3721 tcccctcttt cttcctctag ggtgtcgtta attacccgta ctaaaggttt ggaaaagaaa
     3781 aaagagaccg cctcgtttct ttttcttcgt cgaaaaaggc aataaaaatt tttatcacgt
     3841 ttctttttct tgaaaatttt ttttttgatt tttttctctt tcgatgacct cccattgata
     3901 tttaagttaa taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt
     3961 acaacttttt ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagttttc
     4021 tagaactagt ggatcccccg ggaaaaatgg acaagaagta ctccattggg ctcgatatcg
     4081 gcacaaacag cgtcggttgg gccgtcatta cggacgagta caaggtgccg agcaaaaaat
     4141 tcaaagttct gggcaatacc gatcgccaca gcataaagaa gaacctcatt ggcgccctcc
     4201 tgttcgactc cggggaaacg gccgaagcca cgcggctcaa aagaacagca cggcgcagat
     4261 atacccgcag aaagaatcgg atctgctacc tgcaggagat ctttagtaat gagatggcta
     4321 aggtggatga ctctttcttc cataggctgg aggagtcctt tttggtggag gaggataaaa
     4381 agcacgagcg ccacccaatc tttggcaata tcgtggacga ggtggcgtac catgaaaagt
     4441 acccaaccat atatcatctg aggaagaagc ttgtagacag tactgataag gctgacttgc
     4501 ggttgatcta tctcgcgctg gcgcatatga tcaaatttcg gggacacttc ctcatcgagg
     4561 gggacctgaa cccagacaac agcgatgtcg acaaactctt tatccaactg gttcagactt
     4621 acaatcagct tttcgaagag aacccgatca acgcatccgg agttgacgcc aaagcaatcc
     4681 tgagcgctag gctgtccaaa tcccggcggc tcgaaaacct catcgcacag ctccctgggg
     4741 agaagaagaa cggcctgttt ggtaatctta tcgccctgtc actcgggctg acccccaact
     4801 ttaaatctaa cttcgacctg gccgaagatg ccaagcttca actgagcaaa gacacctacg
     4861 atgatgatct cgacaatctg ctggcccaga tcggcgacca gtacgcagac ctttttttgg
     4921 cggcaaagaa cctgtcagac gccattctgc tgagtgatat tctgcgagtg aacacggaga
     4981 tcaccaaagc tccgctgagc gctagtatga tcaagcgcta tgatgagcac caccaagact
     5041 tgactttgct gaaggccctt gtcagacagc aactgcctga gaagtacaag gaaattttct
     5101 tcgatcagtc taaaaatggc tacgccggat acattgacgg cggagcaagc caggaggaat
     5161 tttacaaatt tattaagccc atcttggaaa aaatggacgg caccgaggag ctgctggtaa
     5221 agcttaacag agaagatctg ttgcgcaaac agcgcacttt cgacaatgga agcatccccc
     5281 accagattca cctgggcgaa ctgcacgcta tcctcaggcg gcaagaggat ttctacccct
     5341 ttttgaaaga taacagggaa aagattgaga aaatcctcac atttcggata ccctactatg
     5401 taggccccct cgcccgggga aattccagat tcgcgtggat gactcgcaaa tcagaagaga
     5461 ccatcactcc ctggaacttc gaggaagtcg tggataaggg ggcctctgcc cagtccttca
     5521 tcgaaaggat gactaacttt gataaaaatc tgcctaacga aaaggtgctt cctaaacact
     5581 ctctgctgta cgagtacttc acagtttata acgagctcac caaggtcaaa tacgtcacag
     5641 aagggatgag aaagccagca ttcctgtctg gagagcagaa gaaagctatc gtggacctcc
     5701 tcttcaagac gaaccggaaa gttaccgtga aacagctcaa agaagactat ttcaaaaaga
     5761 ttgaatgttt cgactctgtt gaaatcagcg gagtggagga tcgcttcaac gcatccctgg
     5821 gaacgtatca cgatctcctg aaaatcatta aagacaagga cttcctggac aatgaggaga
     5881 acgaggacat tcttgaggac attgtcctca cccttacgtt gtttgaagat agggagatga
     5941 ttgaagaacg cttgaaaact tacgctcatc tcttcgacga caaagtcatg aaacagctca
     6001 agaggcgccg atatacagga tgggggcggc tgtcaagaaa actgatcaat gggatccgag
     6061 acaagcagag tggaaagaca atcctggatt ttcttaagtc cgatggattt gccaaccgga
     6121 acttcatgca gttgatccat gatgactctc tcacctttaa ggaggacatc cagaaagcac
     6181 aagtttctgg ccagggggac agtcttcacg agcacatcgc taatcttgca ggtagcccag
     6241 ctatcaaaaa gggaatactg cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg
     6301 gaaggcataa gcccgagaat atcgttatcg agatggcccg agagaaccaa actacccaga
     6361 agggacagaa gaacagtagg gaaaggatga agaggattga agagggtata aaagaactgg
     6421 ggtcccaaat ccttaaggaa cacccagttg aaaacaccca gcttcagaat gagaagctct
     6481 acctgtacta cctgcagaac ggcagggaca tgtacgtgga tcaggaactg gacatcaatc
     6541 ggctctccga ctacgacgtg gatcatatcg tgccccagtc ttttctcaaa gatgattcta
     6601 ttgataataa agtgttgaca agatccgata aaaatagagg gaagagtgat aacgtcccct
     6661 cagaagaagt tgtcaagaaa atgaaaaatt attggcggca gctgctgaac gccaaactga
     6721 tcacacaacg gaagttcgat aatctgacta aggctgaacg aggtggcctg tctgagttgg
     6781 ataaagccgg cttcatcaaa aggcagcttg ttgagacacg ccagatcacc aagcacgtgg
     6841 cccaaattct cgattcacgc atgaacacca agtacgatga aaatgacaaa ctgattcgag
     6901 aggtgaaagt tattactctg aagtctaagc tggtctcaga tttcagaaag gactttcagt
     6961 tttataaggt gagagagatc aacaattacc accatgcgca tgatgcctac ctgaatgcag
     7021 tggtaggcac tgcacttatc aaaaaatatc ccaagcttga atctgaattt gtttacggag
     7081 actataaagt gtacgatgtt aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg
     7141 ccaccgctaa gtacttcttt tacagcaata ttatgaattt tttcaagacc gagattacac
     7201 tggccaatgg agagattcgg aagcgaccac ttatcgaaac aaacggagaa acaggagaaa
     7261 tcgtgtggga caagggtagg gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg
     7321 tgaacatcgt taaaaagacc gaagtacaga ccggaggctt ctccaaggaa agtatcctcc
     7381 cgaaaaggaa cagcgacaag ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg
     7441 gcggattcga ttctcctaca gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag
     7501 ggaagtctaa aaaactcaaa agcgtcaagg aactgctggg catcacaatc atggagcgat
     7561 caagcttcga aaaaaacccc atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa
     7621 aagacctcat cattaagctt cccaagtact ctctctttga gcttgaaaac ggccggaaac
     7681 gaatgctcgc tagtgcgggc gagctgcaga aaggtaacga gctggcactg ccctctaaat
     7741 acgttaattt cttgtatctg gccagccact atgaaaagct caaagggtct cccgaagata
     7801 atgagcagaa gcagctgttc gtggaacaac acaaacacta ccttgatgag atcatcgagc
     7861 aaataagcga attctccaaa agagtgatcc tcgccgacgc taacctcgat aaggtgcttt
     7921 ctgcttacaa taagcacagg gataagccca tcagggagca ggcagaaaac attatccact
     7981 tgtttactct gaccaacttg ggcgcgcctg cagccttcaa gtacttcgac accaccatag
     8041 acagaaagcg gtacacctct acaaaggagg tcctggacgc cacactgatt catcagtcaa
     8101 ttacggggct ctatgaaaca agaatcgacc tctctcagct cggtggagac agcagggctg
     8161 accccaagaa gaagaggaag gtgtgatctc ttctcgagtc atgtaattag ttatgtcacg
     8221 cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     8281 tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat
     8341 atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa
     8401 aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcgcat gcatcaacaa
     8461 cagaggacat atgccctacc tccatgatct actgacgtcc ctgaggctgc aattcatgta
     8521 atggggcagt atccgcggca agtcctagtg caatggcggt tttttaccct cgttctgaag
     8581 aagaggcgac gcgggtgcgg tcatcactaa tgtggaaatt ggtaagactc tcgggcctcc
     8641 gcctttaggc ggtgcttact ctttcataaa ggggctgtta gttatgcaac gatgtgcttc
     8701 agtattacat tttttgcctt caacgccttg attgttctat ttttgctaat aataaatcta
     8761 tttcatcgga ctaaaagtcc attagttgta agcggattta gctcagttgg gagagcgcca
     8821 gactgaagaa aaacttcggt caagtcatct ggaggtcctg tgttcgatcc acagaattcg
     8881 cagatggccg gcatggtccc agcctcctcg ctggcgccgg ctgggcaaca ccttcgggtg
     8941 gcgaatggga cttgagacgg aaagtgaaac gtgatttcat gcgtcatttt gaacattttg
     9001 taaatcttat ttaataatgt gtgcggcaat tcacatttaa tttatgaatg ttttcttaac
     9061 atcgcggcaa ctcaagaaac ggcaggttcg gatcttagct actagagaaa gaggagaaat
     9121 actagatgcg taaaggcgaa gagctgttca ctggtgtcgt ccctattctg gtggaactgg
     9181 atggtgatgt caacggtcat aagttttccg tgcgtggcga gggtgaaggt gacgcaacta
     9241 atggtaaact gacgctgaag ttcatctgta ctactggtaa actgccggtt ccttggccga
     9301 ctctggtaac gacgctgact tatggtgttc agtgctttgc tcgttatccg gaccatatga
     9361 agcagcatga cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa cgcacgattt
     9421 cctttaagga tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa ggcgataccc
     9481 tggtaaaccg cattgagctg aaaggcattg actttaaaga ggacggcaat atcctgggcc
     9541 ataagctgga atacaatttt aacagccaca atgtttacat caccgccgat aaacaaaaaa
     9601 atggcattaa agcgaatttt aaaattcgcc acaacgtgga ggatggcagc gtgcagctgg
     9661 ctgatcacta ccagcaaaac actccaatcg gtgatggtcc tgttctgctg ccagacaatc
     9721 actatctgag cacgcaaagc gttctgtcta aagatccgaa cgagaaacgc gatcatatgg
     9781 ttctgctgga gttcgtaacc gcagcgggca tcacgcatgg tatggatgaa ctgtacaaat
     9841 gaccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     9901 ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt
     9961 ctgcgtttat acgtctcagt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
    10021 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tattttttgt cactattgtt
    10081 atgtaaaatg ccacctctga cagtatggaa cgcaaacttc tgtctagtgg atagctggac
    10141 attccaggca gtgcctctgc cgccggaccc ctctcgtgat tgggtagctg gacatgccct
    10201 tgtaagatat aacaagagcc tgcctgtcta atgatctcac ggcgaaagtc ggggagacag
    10261 cagcggctgc agacattata ccgcaacaac actaaggtga gataactccg tacattgact
    10321 acgcgttcct ctagacctta cttgaccgga tacagtgtct ttgacacgtt tatgggttac
    10381 agcaatcaca tccaagactg gctatgcacg aagcaactct tgagtgttaa aatgttgacc
    10441 cctgtatttg ggatgcgggt agtagatgag tgcaggatgg tacccaattc gccctatagt
    10501 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
    10561 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
    10621 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg
    10681 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
    10741 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
    10801 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
    10861 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
    10921 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
    10981 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
    11041 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
    11101 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
    11161 tctgtgcggt atttcacacc gcatagggta ataactgata taattaaatt gaagctctaa
    11221 tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt tgctggccgc
    11281 atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct accttagcat
    11341 cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct gtagacacca
    11401 catcatccac ggttctatac tgttgaccca atgcatctcc cttgtcatct aaacccacac
    11461 cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct ctttgagcaa
    11521 taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt
    11581 ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg cctctaggtt
    11641 cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg cccaccacac
    11701 cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca gagtactgca
    11761 atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact
    11821 tggcggataa tgcctttagc ggcttaactg tgccctccat ggaaaaatca gtcaagatat
    11881 ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac tccagtaatt
    11941 ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg tgcatgatat
    12001 taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta tatgtagctt
    12061 tcgacatgat ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg gttaagaata
    12121 ctgggcaatt tcatgtttct tcaacactac atatgcgtat atataccaat ctaagtctgt
    12181 gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa
    12241 accgaaatca aaaaaagaat aaaaaaaaaa tgatgaattg aaaaggtggt atggtgcact
    12301 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
    12361 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgac
//
