LOCUS       sequence-322654-       11752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constituive spCas9 and tet-inducible GAL7 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 11752)
  TITLE     Burgess Lab Plasmids
REFERENCE   2  (bases 1 to 11752)
  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_GAL7
FEATURES             Location/Qualifiers
     source          1..11752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3114..3134
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      3150..3397
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3166..3184)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3405..3424)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3406..3424)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3434..3451)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3434..3450)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3443..3465)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3595..4050
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3682..3701)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3892..3913
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4147..4821)
                     /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(4822..5102)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(5117..5136)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5236..5258
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5296..5314)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5382..5486
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5487..5555,5556..6347)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5705..5724)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6518..7106
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7007..7026
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7490..7508
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        7594..7669
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      7682..7744
                     /label=RPR1 terminator
                     /note="transcription terminator for the S. cerevisiae RNase
                     P RNA gene"
     CDS             8895..9518
                     /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(8968..8987)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     10015..10032
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(10185..10190,10191..10208,10209..10215)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10223..10239
                     /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     10228..10250
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10247..10263
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10247..10263
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10282..10302
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10284..10302
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        10320..10718
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     10722..10738
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10722..10738
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             join(10750..11752,1..3101)
                     /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             join(10750..11752,1..3101)
                     /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             join(10750..11752,1..3101)
                     /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"
ORIGIN
        1 tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt
       61 ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat
      121 ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta aagcttaaca
      181 gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc
      241 acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag
      301 ataacaggga aaagattgag aaaatcctca catttcggat accctactat gtaggccccc
      361 tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag accatcactc
      421 cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga
      481 tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt
      541 acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga
      601 gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga
      661 cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag attgaatgtt
      721 tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc
      781 acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca
      841 ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac
      901 gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc
      961 gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga gacaagcaga
     1021 gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc
     1081 agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg
     1141 gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa
     1201 agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata
     1261 agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga
     1321 agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa
     1381 tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact
     1441 acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat cggctctccg
     1501 actacgacgt ggatcatatc gtgccccagt cttttctcaa agatgattct attgataata
     1561 aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag
     1621 ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac
     1681 ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg
     1741 gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc
     1801 tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag
     1861 ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag ttttataagg
     1921 tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca
     1981 ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga gactataaag
     2041 tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta
     2101 agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg
     2161 gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg
     2221 acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag gtgaacatcg
     2281 ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga
     2341 acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg
     2401 attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta
     2461 aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga tcaagcttcg
     2521 aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa aaagacctca
     2581 tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg
     2641 ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt
     2701 tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga
     2761 agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg
     2821 aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca
     2881 ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac ttgtttactc
     2941 tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaagc
     3001 ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc
     3061 tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct gaccccaaga
     3121 agaagaggaa ggtgtgatct cttctcgagt catgtaatta gttatgtcac gcttacattc
     3181 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     3241 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat
     3301 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct
     3361 tgagaaggtt ttgggacgct cgaaggcttt aatttgccaa ttcgccctat agtgagtcgt
     3421 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     3481 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     3541 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcgacgcgc
     3601 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3661 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3721 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3781 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3841 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3901 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3961 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4021 attttaacaa aatattaacg tttacaattt cctgatgcgg tattttctcc ttacgcatct
     4081 gtgcggtatt tcacaccgca ggcaagtgca caaacaatac ttaaataaat actactcagt
     4141 aataacctat ttcttagcat ttttgacgaa atttgctatt ttgttagagt cttttacacc
     4201 atttgtctcc acacctccgc ttacatcaac accaataacg ccatttaatc taagcgcatc
     4261 accaacattt tctggcgtca gtccaccagc taacataaaa tgtaagcttt cggggctctc
     4321 ttgccttcca acccagtcag aaatcgagtt ccaatccaaa agttcacctg tcccacctgc
     4381 ttctgaatca aacaagggaa taaacgaatg aggtttctgt gaagctgcac tgagtagtat
     4441 gttgcagtct tttggaaata cgagtctttt aataactggc aaaccgagga actcttggta
     4501 ttcttgccac gactcatctc catgcagttg gacgatatca atgccgtaat cattgaccag
     4561 agccaaaaca tcctccttag gttgattacg aaacacgcca accaagtatt tcggagtgcc
     4621 tgaactattt ttatatgctt ttacaagact tgaaattttc cttgcaataa ccgggtcaat
     4681 tgttctcttt ctattgggca cacatataat acccagcaag tcagcatcgg aatctagagc
     4741 acattctgcg gcctctgtgc tctgcaagcc gcaaactttc accaatggac cagaactacc
     4801 tgtgaaatta ataacagaca tactccaagc tgcctttgtg tgcttaatca cgtatactca
     4861 cgtgctcaat agtcaccaat gccctccctc ttggccctct ccttttcttt tttcgaccga
     4921 attaattctt aatcggcaaa aaaagaaaag ctccggatca agattgtacg taaggtgaca
     4981 agctattttt caataaagaa tatcttccac tactgccatc tggcgtcata actgcaaagt
     5041 acacatatat tacgatgctg tctattaaat gcttcctata ttatatatat agtaatgtcg
     5101 tttggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     5161 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     5221 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     5281 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5341 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5401 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5461 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5521 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5581 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5641 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5701 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     5761 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5821 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5881 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct ttttttcaca
     5941 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6001 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6061 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6121 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6181 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6241 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6301 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6361 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6421 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6481 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6541 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6601 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6661 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6721 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6781 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6841 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6901 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6961 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7021 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7081 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7141 ccttttgctg gccttttgct cgggggatct gccaattgaa cataacatgg tagttacata
     7201 tactagtaat atggttcggc acacattaaa agtataaaaa ctatctgaat tacgaattac
     7261 atatattggt cataaaaatc aatcaatcat cgtgtgtttt atatgtctct tatctaagta
     7321 taagaatatc catagttaat attcacttac gctacctttt aacctgtaat cattgtcaac
     7381 aggatatgtt aacgacccac attgataaac gctagtattt ctttttcctc ttcttattgg
     7441 ccggctgtct ctatactccc ctatagtctg tttcttttcg tttcgattgt ccctatcagt
     7501 gatagagatg gcgcacatgg tacgctgtgg tgctcgcggc tgggaacgaa actctgggag
     7561 ctgcgattgg caggcctcct gttgacctaa ccagttttag agctagaaat agcaagttaa
     7621 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttctcga
     7681 gccatatcca acttccaatt taatctttct tttttaattt tcacttattt gcgatacaga
     7741 aagaaaaaag cgatagtaac tattgaattt tgtttggatt tggttagatt agatatggtt
     7801 tctctttata tttacatgct aaaaatgggc tacaccagag atacataatt agatatatat
     7861 acgccagtac accttatcgg cccaagcctt gtcccaaggc agcgttttgt tcttggaaac
     7921 gctgccctac acgttcgcta tgcttcaaga acttttctga gcacttcatg atgcatgttt
     7981 gttccttatt ggttagcttt gatgttgtga agtcattgac acagtctgtg aaacatcttt
     8041 ctaccagatt agagtacaaa cgcatgaaat ccttcatttg cttttgttcc actacttttt
     8101 ggaactcttg ttgttctttc ctgcaggtag tcgtgcaatg tatgacttta agatttgtga
     8161 gcaggaagaa aagggagaat cttctaacga taaacccttg aaaaactggg tagactacgc
     8221 tatgttgagt tgctacgcag gctgcacaat tacacgagaa tgctcccgcc taggatttaa
     8281 ggctaaggga cgtgcaatgc agacgacaga tctaaatgac cgtgtcggtg aagtgttcgc
     8341 caaacttttc ggttaacaca tgcagtgatg cacgcgcgat ggtgctaagt tacatatata
     8401 tatatatata tatatatata tatatatagc catagtgatg tctaagtaac ctttatggta
     8461 tatttcttaa tgtggaaaga tactagcgcg cgcacccaca cacaagcttc gtcttttctt
     8521 gaagaaaaga ggaagctcgc taaatgggat tccactttcc gttccctgcc agctgatgga
     8581 aaaaggttag tggaacgatg aagaataaaa agagagatcc actgaggtga aatttcagct
     8641 gacagcgagt ttcatgatcg tgatgaacaa tggtaacgag ttgtggctgt tgccagggag
     8701 ggtggttctc aacttttaat gtatggccaa atcgctactt gggtttgtta tataacaaag
     8761 aagaaataat gaactgattc tcttcctcct tcttgtcctt tcttaattct gttgtaatta
     8821 ccttcctttg taattttttt tgtaattatt cttcttaata atccaaacaa acacacatat
     8881 tacaatatat taaaatgtct agattagata aaagtaaagt gattaacagc gcattagagc
     8941 tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa actcgcccag aagctaggtg
     9001 tagagcagcc tacattgtat tggcatgtaa aaaataagcg ggctttgctc gacgccttag
     9061 ccattgagat gttagatagg caccatactc acttttgccc tttagaaggg gaaagctggc
     9121 aagatttttt acgtaataac gctaaaagtt ttagatgtgc tttactaagt catcgcgatg
     9181 gagcaaaagt acatttaggt acacggccta cagaaaaaca gtatgaaact ctcgaaaatc
     9241 aattagcctt tttatgccaa caaggttttt cactagagaa tgcattatat gcactcagcg
     9301 ctgtggggca ttttacttta ggttgcgtat tggaagatca agagcatcaa gtcgctaaag
     9361 aagaaaggga aacacctact actgatagta tgccgccatt attacgacaa gctatcgaat
     9421 tatttgatca ccaaggtgca gagccagcct tcttattcgg ccttgaattg atcatatgcg
     9481 gattagaaaa acaacttaaa tgtgaaagtg ggtcttaagt ctgaagaatg aatgatttga
     9541 tgatttcttt ttccctccat ttttcttact gaatatatca atgatataga cttgtatagt
     9601 ttattatttc aaattaagta gctatatata gtcaagataa cgtttgtttg acacgattac
     9661 attattcgtc gacatctttt ttcagcctgt cgtggtagca atttgaggag tattattaat
     9721 tgaataggtt cattttgcgc tcgcataaac agttttcgtc agggacagta tgttggaatg
     9781 agtggtaatt aatggtgaca tgacatgtta tagcaataac cttgatgttt acatcgtagt
     9841 ttaatgtaca ccccgcgaat tcgttcaagt aggagtgcac caattgcaaa gggaaaagct
     9901 gaatgggcag ttcgaaccgc ggtctttcct gcgttatccc ctgattctgt ggataaccgt
     9961 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
    10021 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
    10081 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
    10141 aacgcaatta atgtgagtta cctcactcat taggcacccc aggctttaca ctttatgctt
    10201 ccggctccta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
    10261 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctc
    10321 atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca
    10381 tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc
    10441 tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt
    10501 tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat
    10561 tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt taataaacgg
    10621 tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc
    10681 ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttctagaact agtggatccc
    10741 ccgggaaaaa tggacaagaa gtactccatt gggctcgata tcggcacaaa cagcgtcggt
    10801 tgggccgtca ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt tctgggcaat
    10861 accgatcgcc acagcataaa gaagaacctc attggcgccc tcctgttcga ctccggggag
    10921 acggccgaag ccacgcggct caaaagaaca gcacggcgca gatatacccg cagaaagaat
    10981 cggatctgct acctgcagga gatctttagt aatgagatgg ctaaggtgga tgactctttc
    11041 ttccataggc tggaggagtc ctttttggtg gaggaggata aaaagcacga gcgccaccca
    11101 atctttggca atatcgtgga cgaggtggcg taccatgaaa agtacccaac catatatcat
    11161 ctgaggaaga agcttgtaga cagtactgat aaggctgact tgcggttgat ctatctcgcg
    11221 ctggcgcata tgatcaaatt tcggggacac ttcctcatcg agggggacct gaacccagac
    11281 aacagcgatg tcgacaaact ctttatccaa ctggttcaga cttacaatca gcttttcgaa
    11341 gagaacccga tcaacgcatc cggagttgac gccaaagcaa tcctgagcgc taggctgtcc
    11401 aaatcccggc ggctcgaaaa cctcatcgca cagctccctg gggagaagaa gaacggcctg
    11461 tttggtaatc ttatcgccct gtcactcggg ctgaccccca actttaaatc taacttcgac
    11521 ctggccgaag atgccaagct tcaactgagc aaagacacct acgatgatga tctcgacaat
    11581 ctgctggccc agatcggcga ccagtacgca gacctttttt tggcggcaaa gaacctgtca
    11641 gacgccattc tgctgagtga tattctgcga gtgaacacgg agatcaccaa agctccgctg
    11701 agcgctagta tgatcaagcg ctatgatgag caccaccaag acttgacttt gc
//
