LOCUS       sequence-329133-       11755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constituive spCas9 and tet-inducible APL1-targeting 
            sgRNA expression for double stranded break formation in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11755)
  TITLE     Burgess Lab Plasmids
REFERENCE   2  (bases 1 to 11755)
  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"
COMMENT     Sequence Label: pNSU299_APL3
FEATURES             Location/Qualifiers
     source          1..11755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1037..1660
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(1110..1129)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     2157..2174
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2291..2312
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2327..2332,2333..2350,2351..2357)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2365..2381
                     /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     2370..2392
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2389..2405
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2389..2405
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2424..2444
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2426..2444
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2462..2860
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     2864..2880
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2864..2880
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2892..6995
                     /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             2892..6995
                     /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             2892..6995
                     /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             7008..7028
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7044..7291
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7060..7078)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(7309..7328)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7310..7328)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7338..7355)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7338..7354)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7347..7369)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7495..7950
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7582..7601)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7792..7813
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(8053..8727)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(8728..9009)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(9030..9049)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9149..9171
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9209..9227)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9295..9399
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9400..9468,9469..10260)
                     /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(9618..9637)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10431..11019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10920..10939
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    11403..11421
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        11507..11582
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      11595..11657
                     /label=RPR1 terminator
                     /note="transcription terminator for the S. cerevisiae RNase
                     P RNA gene"
ORIGIN
        1 acataattag atatatatac gccagtacac cttatcggcc caagccttgt cccaaggcag
       61 cgttttgttc ttggaaacgc tgccctacac gttcgctatg cttcaagaac ttttctgagc
      121 acttcatgat gcatgtttgt tccttattgg ttagctttga tgttgtgaag tcattgacac
      181 agtctgtgaa acatctttct accagattag agtacaaacg catgaaatcc ttcatttgct
      241 tttgttccac tactttttgg aactcttgtt gttctttcct gcaggtagtc gtgcaatgta
      301 tgactttaag atttgtgagc aggaagaaaa gggagaatct tctaacgata aacccttgaa
      361 aaactgggta gactacgcta tgttgagttg ctacgcaggc tgcacaatta cacgagaatg
      421 ctcccgccta ggatttaagg ctaagggacg tgcaatgcag acgacagatc taaatgaccg
      481 tgtcggtgaa gtgttcgcca aacttttcgg ttaacacatg cagtgatgca cgcgcgatgg
      541 tgctaagtta catatatata tatatgtata gccatagtga tgtctaagta acctttatgg
      601 tatatttctt aatgtggaaa gatactagcg cgcgcaccca cacacaagct tcgtcttttc
      661 ttgaagaaaa gaggaagctc gctaaatggg attccacttt ccgttccctg ccagctgatg
      721 gaaaaaggtt agtggaacga tgaagaataa aaagagagat ccactgaggt gaaatttcag
      781 ctgacagcga gtttcatgat cgtgatgaac aatggtaacg agttgtggct gttgccaggg
      841 agggtggttc tcaactttta atgtatggcc aaatcgctac ttgggtttgt tatataacaa
      901 agaagaaata atgaactgat tctcttcctc cttcttgtcc tttcttaatt ctgttgtaat
      961 taccttcctt tgtaattttt tttgtaatta ttcttcttaa taatccaaac aaacacacat
     1021 attacaatat attaaaatgt ctagattaga taaaagtaaa gtgattaaca gcgcattaga
     1081 gctgcttaat gaggtcggaa tcgaaggttt aacaacccgt aaactcgccc agaagctagg
     1141 tgtagagcag cctacattgt attggcatgt aaaaaataag cgggctttgc tcgacgcctt
     1201 agccattgag atgttagata ggcaccatac tcacttttgc cctttagaag gggaaagctg
     1261 gcaagatttt ttacgtaata acgctaaaag ttttagatgt gctttactaa gtcatcgcga
     1321 tggagcaaaa gtacatttag gtacacggcc tacagaaaaa cagtatgaaa ctctcgaaaa
     1381 tcaattagcc tttttatgcc aacaaggttt ttcactagag aatgcattat atgcactcag
     1441 cgctgtgggg cattttactt taggttgcgt attggaagat caagagcatc aagtcgctaa
     1501 agaagaaagg gaaacaccta ctactgatag tatgccgcca ttattacgac aagctatcga
     1561 attatttgat caccaaggtg cagagccagc cttcttattc ggccttgaat tgatcatatg
     1621 cggattagaa aaacaactta aatgtgaaag tgggtcttaa gtctgaagaa tgaatgattt
     1681 gatgatttct ttttccctcc atttttctta ctgaatatat caatgatata gacttgtata
     1741 gtttattatt tcaaattaag tagctatata tagtcaagat aacgtttgtt tgacacgatt
     1801 acattattcg tcgacatctt ttttcagcct gtcgtggtag caatttgagg agtattatta
     1861 attgaatagg ttcattttgc gctcgcataa acagttttcg tcagggacag tatgttggaa
     1921 tgagtggtaa ttaatggtga catgacatgt tatagcaata accttgatgt ttacatcgta
     1981 gtttaatgta caccccgcga attcgttcaa gtaggagtgc accaattgca aagggaaaag
     2041 ctgaatgggc agttcgaacc gcggtctttc ctgcgttatc ccctgattct gtggataacc
     2101 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     2161 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     2221 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     2281 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     2341 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     2401 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
     2461 tcatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg
     2521 catcgccgta ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc
     2581 tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg
     2641 tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa
     2701 attttttttt tgattttttt ctctttcgat gacctcccat tgatatttaa gttaataaac
     2761 ggtcttcaat ttctcaagtt tcagtttcat ttttcttgtt ctattacaac tttttttact
     2821 tcttgctcat tagaaagaaa gcatagcaat ctaatctaag ttttctagaa ctagtggatc
     2881 ccccgggaaa aatggacaag aagtactcca ttgggctcga tatcggcaca aacagcgtcg
     2941 gttgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa gttctgggca
     3001 ataccgatcg ccacagcata aagaagaacc tcattggcgc cctcctgttc gactccgggg
     3061 agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc cgcagaaaga
     3121 atcggatctg ctacctgcag gagatcttta gtaatgagat ggctaaggtg gatgactctt
     3181 tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagcac gagcgccacc
     3241 caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca accatatatc
     3301 atctgaggaa gaagcttgta gacagtactg ataaggctga cttgcggttg atctatctcg
     3361 cgctggcgca tatgatcaaa tttcggggac acttcctcat cgagggggac ctgaacccag
     3421 acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat cagcttttcg
     3481 aagagaaccc gatcaacgca tccggagttg acgccaaagc aatcctgagc gctaggctgt
     3541 ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag aagaacggcc
     3601 tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa tctaacttcg
     3661 acctggccga agatgccaag cttcaactga gcaaagacac ctacgatgat gatctcgaca
     3721 atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca aagaacctgt
     3781 cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc aaagctccgc
     3841 tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact ttgctgaagg
     3901 cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat cagtctaaaa
     3961 atggctacgc cggatacatt gacggcggag caagccagga ggaattttac aaatttatta
     4021 agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt aacagagaag
     4081 atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag attcacctgg
     4141 gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg aaagataaca
     4201 gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc cccctcgccc
     4261 ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc actccctgga
     4321 acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa aggatgacta
     4381 actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg ctgtacgagt
     4441 acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg atgagaaagc
     4501 cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc aagacgaacc
     4561 ggaaagttac cgtgaaacag ctcaaagaag actatttcaa aaagattgaa tgtttcgact
     4621 ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg tatcacgatc
     4681 tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag gacattcttg
     4741 aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa gaacgcttga
     4801 aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagagg cgccgatata
     4861 caggatgggg gcggctgtca agaaaactga tcaatgggat ccgagacaag cagagtggaa
     4921 agacaatcct ggattttctt aagtccgatg gatttgccaa ccggaacttc atgcagttga
     4981 tccatgatga ctctctcacc tttaaggagg acatccagaa agcacaagtt tctggccagg
     5041 gggacagtct tcacgagcac atcgctaatc ttgcaggtag cccagctatc aaaaagggaa
     5101 tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg cataagcccg
     5161 agaatatcgt tatcgagatg gcccgagaga accaaactac ccagaaggga cagaagaaca
     5221 gtagggaaag gatgaagagg attgaagagg gtataaaaga actggggtcc caaatcctta
     5281 aggaacaccc agttgaaaac acccagcttc agaatgagaa gctctacctg tactacctgc
     5341 agaacggcag ggacatgtac gtggatcagg aactggacat caatcggctc tccgactacg
     5401 acgtggatca tatcgtgccc cagtcttttc tcaaagatga ttctattgat aataaagtgt
     5461 tgacaagatc cgataaaaat agagggaaga gtgataacgt cccctcagaa gaagttgtca
     5521 agaaaatgaa aaattattgg cggcagctgc tgaacgccaa actgatcaca caacggaagt
     5581 tcgataatct gactaaggct gaacgaggtg gcctgtctga gttggataaa gccggcttca
     5641 tcaaaaggca gcttgttgag acacgccaga tcaccaagca cgtggcccaa attctcgatt
     5701 cacgcatgaa caccaagtac gatgaaaatg acaaactgat tcgagaggtg aaagttatta
     5761 ctctgaagtc taagctggtc tcagatttca gaaaggactt tcagttttat aaggtgagag
     5821 agatcaacaa ttaccaccat gcgcatgatg cctacctgaa tgcagtggta ggcactgcac
     5881 ttatcaaaaa atatcccaag cttgaatctg aatttgttta cggagactat aaagtgtacg
     5941 atgttaggaa aatgatcgca aagtctgagc aggaaatagg caaggccacc gctaagtact
     6001 tcttttacag caatattatg aattttttca agaccgagat tacactggcc aatggagaga
     6061 ttcggaagcg accacttatc gaaacaaacg gagaaacagg agaaatcgtg tgggacaagg
     6121 gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc gcaggtgaac atcgttaaaa
     6181 agaccgaagt acagaccgga ggcttctcca aggaaagtat cctcccgaaa aggaacagcg
     6241 acaagctgat cgcacgcaaa aaagattggg accccaagaa atacggcgga ttcgattctc
     6301 ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag tctaaaaaac
     6361 tcaaaagcgt caaggaactg ctgggcatca caatcatgga gcgatcaagc ttcgaaaaaa
     6421 accccatcga ctttctcgag gcgaaaggat ataaagaggt caaaaaagac ctcatcatta
     6481 agcttcccaa gtactctctc tttgagcttg aaaacggccg gaaacgaatg ctcgctagtg
     6541 cgggcgagct gcagaaaggt aacgagctgg cactgccctc taaatacgtt aatttcttgt
     6601 atctggccag ccactatgaa aagctcaaag ggtctcccga agataatgag cagaagcagc
     6661 tgttcgtgga acaacacaaa cactaccttg atgagatcat cgagcaaata agcgaattct
     6721 ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt gctttctgct tacaataagc
     6781 acagggataa gcccatcagg gagcaggcag aaaacattat ccacttgttt actctgacca
     6841 acttgggcgc gcctgcagcc ttcaagtact tcgacaccac catagacaga aagcggtaca
     6901 cctctacaaa ggaggtcctg gacgccacac tgattcatca gtcaattacg gggctctatg
     6961 aaacaagaat cgacctctct cagctcggtg gagacagcag ggctgacccc aagaagaaga
     7021 ggaaggtgtg atctcttctc gagtcatgta attagttatg tcacgcttac attcacgccc
     7081 tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
     7141 tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct
     7201 tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa
     7261 ggttttggga cgctcgaagg ctttaatttg cggccggtac ccaattcgcc ctatagtgag
     7321 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     7381 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     7441 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc
     7501 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     7561 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     7621 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     7681 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     7741 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     7801 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     7861 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     7921 attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     7981 gtgcggtatt tcacaccgca tagatcggca agtgcacaaa caatacttaa ataaatacta
     8041 ctcagtaata acctatttct tagcattttt gacgaaattt gctattttgt tagagtcttt
     8101 tacaccattt gtctccacac ctccgcttac atcaacacca ataacgccat ttaatctaag
     8161 cgcatcacca acattttctg gcgtcagtcc accagctaac ataaaatgta agctttcggg
     8221 gctctcttgc cttccaaccc agtcagaaat cgagttccaa tccaaaagtt cacctgtccc
     8281 acctgcttct gaatcaaaca agggaataaa cgaatgaggt ttctgtgaag ctgcactgag
     8341 tagtatgttg cagtcttttg gaaatacgag tcttttaata actggcaaac cgaggaactc
     8401 ttggtattct tgccacgact catctccatg cagttggacg atatcaatgc cgtaatcatt
     8461 gaccagagcc aaaacatcct ccttaggttg attacgaaac acgccaacca agtatttcgg
     8521 agtgcctgaa ctatttttat atgcttttac aagacttgaa attttccttg caataaccgg
     8581 gtcaattgtt ctctttctat tgggcacaca tataataccc agcaagtcag catcggaatc
     8641 tagagcacat tctgcggcct ctgtgctctg caagccgcaa actttcacca atggaccaga
     8701 actacctgtg aaattaataa cagacatact ccaagctgcc tttgtgtgct taatcacgta
     8761 tactcacgtg ctcaatagtc accaatgccc tccctcttgg ccctctcctt ttcttttttc
     8821 gaccgaatta attcttaatc ggcaaaaaaa gaaaagctcc ggatcaagat tgtacgtaag
     8881 gtgacaagct atttttcaat aaagaatatc ttccactact gccatctggc gtcataactg
     8941 caaagtacac atatattacg atgctgttct attaaatgct tcctatatta tatatatagt
     9001 aatgtcgttg atctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     9061 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     9121 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     9181 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     9241 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     9301 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     9361 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     9421 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     9481 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     9541 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     9601 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     9661 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     9721 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     9781 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     9841 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     9901 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     9961 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
    10021 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
    10081 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
    10141 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
    10201 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
    10261 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
    10321 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
    10381 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
    10441 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
    10501 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
    10561 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
    10621 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
    10681 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
    10741 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
    10801 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
    10861 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
    10921 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
    10981 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
    11041 ttacggttcc tggccttttg ctggcctttt gctcggggga tctgccaatt gaacataaca
    11101 tggtagttac atatactagt aatatggttc ggcacacatt aaaagtataa aaactatctg
    11161 aattacgaat tacatatatt ggtcataaaa atcaatcaat catcgtgtgt tttatatgtc
    11221 tcttatctaa gtataagaat atccatagtt aatattcact tacgctacct tttaacctgt
    11281 aatcattgtc aacaggatat gttaacgacc cacattgata aacgctagta tttctttttc
    11341 ctcttcttat tggccggctg tctctatact cccctatagt ctgtttcttt tcgtttcgat
    11401 tgtccctatc agtgatagag atggcgcaca tggtacgctg tggtgctcgc ggctgggaac
    11461 gaaactctgg gagctgcgat tggcaggatc ctcgctccaa attggagttt tagagctaga
    11521 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
    11581 gcttttttct cgagccatat ccaacttcca atttaatctt tcttttttaa ttttcactta
    11641 tttgcgatac agaaagaaaa aagcgatagt aactattgaa ttttgtttgg atttggttag
    11701 attagatatg gtttctcttt atatttacat gctaaaaatg ggctacacca gagat
//
