LOCUS       sequence-329603-       11755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constituive spCas9 expression and tet-inducible 
            expression of sgRNA targeting intergenic site near HXT6 (within 
            YDR344C) 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_YDR344C
FEATURES             Location/Qualifiers
     source          1..11755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             464..1087
                     /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(537..556)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     1584..1601
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1718..1739
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1754..1759,1760..1777,1778..1784)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1792..1808
                     /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     1797..1819
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1816..1832
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1816..1832
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1851..1871
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1853..1871
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1889..2287
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     2291..2307
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2291..2307
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2319..6422
                     /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             2319..6422
                     /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             2319..6422
                     /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             6435..6455
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6471..6718
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6487..6505)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6736..6755)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6737..6755)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6765..6782)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6765..6781)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6774..6796)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6922..7377
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7009..7028)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7219..7240
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7480..8154)
                     /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(8155..8436)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(8457..8476)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8576..8598
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8636..8654)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8722..8826
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8827..8895,8896..9687)
                     /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(9045..9064)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9858..10446
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10347..10366
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    10830..10848
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        10934..11009
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      11022..11084
                     /label=RPR1 terminator
                     /note="transcription terminator for the S. cerevisiae RNase
                     P RNA gene"
ORIGIN
        1 atagtgatgt ctaagtaacc tttatggtat atttcttaat gtggaaagat actagcgcgc
       61 gcacccacac acaagcttcg tcttttcttg aagaaaagag gaagctcgct aaatgggatt
      121 ccactttccg ttccctgcca gctgatggaa aaaggttagt ggaacgatga agaataaaaa
      181 gagagatcca ctgaggtgaa atttcagctg acagcgagtt tcatgatcgt gatgaacaat
      241 ggtaacgagt tgtggctgtt gccagggagg gtggttctca acttttaatg tatggccaaa
      301 tcgctacttg ggtttgttat ataacaaaga agaaataatg aactgattct cttcctcctt
      361 cttgtccttt cttaattctg ttgtaattac cttcctttgt aatttttttt gtaattattc
      421 ttcttaataa tccaaacaaa cacacatatt acaatatatt aaaatgtcta gattagataa
      481 aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac
      541 aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcatgtaaa
      601 aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc accatactca
      661 cttttgccct ttagaagggg aaagctggca agatttttta cgtaataacg ctaaaagttt
      721 tagatgtgct ttactaagtc atcgcgatgg agcaaaagta catttaggta cacggcctac
      781 agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac aaggtttttc
      841 actagagaat gcattatatg cactcagcgc tgtggggcat tttactttag gttgcgtatt
      901 ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta ctgatagtat
      961 gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag agccagcctt
     1021 cttattcggc cttgaattga tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg
     1081 gtcttaagtc tgaagaatga atgatttgat gatttctttt tccctccatt tttcttactg
     1141 aatatatcaa tgatatagac ttgtatagtt tattatttca aattaagtag ctatatatag
     1201 tcaagataac gtttgtttga cacgattaca ttattcgtcg acatcttttt tcagcctgtc
     1261 gtggtagcaa tttgaggagt attattaatt gaataggttc attttgcgct cgcataaaca
     1321 gttttcgtca gggacagtat gttggaatga gtggtaatta atggtgacat gacatgttat
     1381 agcaataacc ttgatgttta catcgtagtt taatgtacac cccgcgaatt cgttcaagta
     1441 ggagtgcacc aattgcaaag ggaaaagctg aatgggcagt tcgaaccgcg gtctttcctg
     1501 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     1561 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     1621 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     1681 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     1741 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     1801 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
     1861 ctcactaaag ggaacaaaag ctggagctca tagcttcaaa atgtttctac tcctttttta
     1921 ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca
     1981 gcatactaaa tttcccctct ttcttcctct agggtgtcgt taattacccg tactaaaggt
     2041 ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa
     2101 tttttatcac gtttcttttt cttgaaaatt ttttttttga tttttttctc tttcgatgac
     2161 ctcccattga tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt
     2221 tcttgttcta ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta
     2281 atctaagttt tctagaacta gtggatcccc cgggaaaaat ggacaagaag tactccattg
     2341 ggctcgatat cggcacaaac agcgtcggtt gggccgtcat tacggacgag tacaaggtgc
     2401 cgagcaaaaa attcaaagtt ctgggcaata ccgatcgcca cagcataaag aagaacctca
     2461 ttggcgccct cctgttcgac tccggggaga cggccgaagc cacgcggctc aaaagaacag
     2521 cacggcgcag atatacccgc agaaagaatc ggatctgcta cctgcaggag atctttagta
     2581 atgagatggc taaggtggat gactctttct tccataggct ggaggagtcc tttttggtgg
     2641 aggaggataa aaagcacgag cgccacccaa tctttggcaa tatcgtggac gaggtggcgt
     2701 accatgaaaa gtacccaacc atatatcatc tgaggaagaa gcttgtagac agtactgata
     2761 aggctgactt gcggttgatc tatctcgcgc tggcgcatat gatcaaattt cggggacact
     2821 tcctcatcga gggggacctg aacccagaca acagcgatgt cgacaaactc tttatccaac
     2881 tggttcagac ttacaatcag cttttcgaag agaacccgat caacgcatcc ggagttgacg
     2941 ccaaagcaat cctgagcgct aggctgtcca aatcccggcg gctcgaaaac ctcatcgcac
     3001 agctccctgg ggagaagaag aacggcctgt ttggtaatct tatcgccctg tcactcgggc
     3061 tgacccccaa ctttaaatct aacttcgacc tggccgaaga tgccaagctt caactgagca
     3121 aagacaccta cgatgatgat ctcgacaatc tgctggccca gatcggcgac cagtacgcag
     3181 accttttttt ggcggcaaag aacctgtcag acgccattct gctgagtgat attctgcgag
     3241 tgaacacgga gatcaccaaa gctccgctga gcgctagtat gatcaagcgc tatgatgagc
     3301 accaccaaga cttgactttg ctgaaggccc ttgtcagaca gcaactgcct gagaagtaca
     3361 aggaaatttt cttcgatcag tctaaaaatg gctacgccgg atacattgac ggcggagcaa
     3421 gccaggagga attttacaaa tttattaagc ccatcttgga aaaaatggac ggcaccgagg
     3481 agctgctggt aaagcttaac agagaagatc tgttgcgcaa acagcgcact ttcgacaatg
     3541 gaagcatccc ccaccagatt cacctgggcg aactgcacgc tatcctcagg cggcaagagg
     3601 atttctaccc ctttttgaaa gataacaggg aaaagattga gaaaatcctc acatttcgga
     3661 taccctacta tgtaggcccc ctcgcccggg gaaattccag attcgcgtgg atgactcgca
     3721 aatcagaaga gaccatcact ccctggaact tcgaggaagt cgtggataag ggggcctctg
     3781 cccagtcctt catcgaaagg atgactaact ttgataaaaa tctgcctaac gaaaaggtgc
     3841 ttcctaaaca ctctctgctg tacgagtact tcacagttta taacgagctc accaaggtca
     3901 aatacgtcac agaagggatg agaaagccag cattcctgtc tggagagcag aagaaagcta
     3961 tcgtggacct cctcttcaag acgaaccgga aagttaccgt gaaacagctc aaagaagact
     4021 atttcaaaaa gattgaatgt ttcgactctg ttgaaatcag cggagtggag gatcgcttca
     4081 acgcatccct gggaacgtat cacgatctcc tgaaaatcat taaagacaag gacttcctgg
     4141 acaatgagga gaacgaggac attcttgagg acattgtcct cacccttacg ttgtttgaag
     4201 atagggagat gattgaagaa cgcttgaaaa cttacgctca tctcttcgac gacaaagtca
     4261 tgaaacagct caagaggcgc cgatatacag gatgggggcg gctgtcaaga aaactgatca
     4321 atgggatccg agacaagcag agtggaaaga caatcctgga ttttcttaag tccgatggat
     4381 ttgccaaccg gaacttcatg cagttgatcc atgatgactc tctcaccttt aaggaggaca
     4441 tccagaaagc acaagtttct ggccaggggg acagtcttca cgagcacatc gctaatcttg
     4501 caggtagccc agctatcaaa aagggaatac tgcagaccgt taaggtcgtg gatgaactcg
     4561 tcaaagtaat gggaaggcat aagcccgaga atatcgttat cgagatggcc cgagagaacc
     4621 aaactaccca gaagggacag aagaacagta gggaaaggat gaagaggatt gaagagggta
     4681 taaaagaact ggggtcccaa atccttaagg aacacccagt tgaaaacacc cagcttcaga
     4741 atgagaagct ctacctgtac tacctgcaga acggcaggga catgtacgtg gatcaggaac
     4801 tggacatcaa tcggctctcc gactacgacg tggatcatat cgtgccccag tcttttctca
     4861 aagatgattc tattgataat aaagtgttga caagatccga taaaaataga gggaagagtg
     4921 ataacgtccc ctcagaagaa gttgtcaaga aaatgaaaaa ttattggcgg cagctgctga
     4981 acgccaaact gatcacacaa cggaagttcg ataatctgac taaggctgaa cgaggtggcc
     5041 tgtctgagtt ggataaagcc ggcttcatca aaaggcagct tgttgagaca cgccagatca
     5101 ccaagcacgt ggcccaaatt ctcgattcac gcatgaacac caagtacgat gaaaatgaca
     5161 aactgattcg agaggtgaaa gttattactc tgaagtctaa gctggtctca gatttcagaa
     5221 aggactttca gttttataag gtgagagaga tcaacaatta ccaccatgcg catgatgcct
     5281 acctgaatgc agtggtaggc actgcactta tcaaaaaata tcccaagctt gaatctgaat
     5341 ttgtttacgg agactataaa gtgtacgatg ttaggaaaat gatcgcaaag tctgagcagg
     5401 aaataggcaa ggccaccgct aagtacttct tttacagcaa tattatgaat tttttcaaga
     5461 ccgagattac actggccaat ggagagattc ggaagcgacc acttatcgaa acaaacggag
     5521 aaacaggaga aatcgtgtgg gacaagggta gggatttcgc gacagtccgg aaggtcctgt
     5581 ccatgccgca ggtgaacatc gttaaaaaga ccgaagtaca gaccggaggc ttctccaagg
     5641 aaagtatcct cccgaaaagg aacagcgaca agctgatcgc acgcaaaaaa gattgggacc
     5701 ccaagaaata cggcggattc gattctccta cagtcgctta cagtgtactg gttgtggcca
     5761 aagtggagaa agggaagtct aaaaaactca aaagcgtcaa ggaactgctg ggcatcacaa
     5821 tcatggagcg atcaagcttc gaaaaaaacc ccatcgactt tctcgaggcg aaaggatata
     5881 aagaggtcaa aaaagacctc atcattaagc ttcccaagta ctctctcttt gagcttgaaa
     5941 acggccggaa acgaatgctc gctagtgcgg gcgagctgca gaaaggtaac gagctggcac
     6001 tgccctctaa atacgttaat ttcttgtatc tggccagcca ctatgaaaag ctcaaagggt
     6061 ctcccgaaga taatgagcag aagcagctgt tcgtggaaca acacaaacac taccttgatg
     6121 agatcatcga gcaaataagc gaattctcca aaagagtgat cctcgccgac gctaacctcg
     6181 ataaggtgct ttctgcttac aataagcaca gggataagcc catcagggag caggcagaaa
     6241 acattatcca cttgtttact ctgaccaact tgggcgcgcc tgcagccttc aagtacttcg
     6301 acaccaccat agacagaaag cggtacacct ctacaaagga ggtcctggac gccacactga
     6361 ttcatcagtc aattacgggg ctctatgaaa caagaatcga cctctctcag ctcggtggag
     6421 acagcagggc tgaccccaag aagaagagga aggtgtgatc tcttctcgag tcatgtaatt
     6481 agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg
     6541 agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag
     6601 aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa
     6661 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg
     6721 ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     6781 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     6841 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     6901 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6961 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     7021 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     7081 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     7141 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     7201 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     7261 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     7321 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct
     7381 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag atcggcaagt
     7441 gcacaaacaa tacttaaata aatactactc agtaataacc tatttcttag catttttgac
     7501 gaaatttgct attttgttag agtcttttac accatttgtc tccacacctc cgcttacatc
     7561 aacaccaata acgccattta atctaagcgc atcaccaaca ttttctggcg tcagtccacc
     7621 agctaacata aaatgtaagc tttcggggct ctcttgcctt ccaacccagt cagaaatcga
     7681 gttccaatcc aaaagttcac ctgtcccacc tgcttctgaa tcaaacaagg gaataaacga
     7741 atgaggtttc tgtgaagctg cactgagtag tatgttgcag tcttttggaa atacgagtct
     7801 tttaataact ggcaaaccga ggaactcttg gtattcttgc cacgactcat ctccatgcag
     7861 ttggacgata tcaatgccgt aatcattgac cagagccaaa acatcctcct taggttgatt
     7921 acgaaacacg ccaaccaagt atttcggagt gcctgaacta tttttatatg cttttacaag
     7981 acttgaaatt ttccttgcaa taaccgggtc aattgttctc tttctattgg gcacacatat
     8041 aatacccagc aagtcagcat cggaatctag agcacattct gcggcctctg tgctctgcaa
     8101 gccgcaaact ttcaccaatg gaccagaact acctgtgaaa ttaataacag acatactcca
     8161 agctgccttt gtgtgcttaa tcacgtatac tcacgtgctc aatagtcacc aatgccctcc
     8221 ctcttggccc tctccttttc ttttttcgac cgaattaatt cttaatcggc aaaaaaagaa
     8281 aagctccgga tcaagattgt acgtaaggtg acaagctatt tttcaataaa gaatatcttc
     8341 cactactgcc atctggcgtc ataactgcaa agtacacata tattacgatg ctgttctatt
     8401 aaatgcttcc tatattatat atatagtaat gtcgttgatc tatggtgcac tctcagtaca
     8461 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     8521 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     8581 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     8641 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     8701 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     8761 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     8821 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     8881 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     8941 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     9001 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     9061 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     9121 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     9181 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     9241 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     9301 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     9361 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     9421 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     9481 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     9541 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     9601 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     9661 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     9721 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     9781 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     9841 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     9901 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     9961 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
    10021 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
    10081 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
    10141 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
    10201 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
    10261 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
    10321 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
    10381 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
    10441 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
    10501 cgggggatct gccaattgaa cataacatgg tagttacata tactagtaat atggttcggc
    10561 acacattaaa agtataaaaa ctatctgaat tacgaattac atatattggt cataaaaatc
    10621 aatcaatcat cgtgtgtttt atatgtctct tatctaagta taagaatatc catagttaat
    10681 attcacttac gctacctttt aacctgtaat cattgtcaac aggatatgtt aacgacccac
    10741 attgataaac gctagtattt ctttttcctc ttcttattgg ccggctgtct ctatactccc
    10801 ctatagtctg tttcttttcg tttcgattgt ccctatcagt gatagagatg gcgcacatgg
    10861 tacgctgtgg tgctcgcggc tgggaacgaa actctgggag ctgcgattgg cagtgaacat
    10921 gttgcaacaa ctggttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
    10981 aacttgaaaa agtggcaccg agtcggtgct tttttctcga gccatatcca acttccaatt
    11041 taatctttct tttttaattt tcacttattt gcgatacaga aagaaaaaag cgatagtaac
    11101 tattgaattt tgtttggatt tggttagatt agatatggtt tctctttata tttacatgct
    11161 aaaaatgggc tacaccagag atacataatt agatatatat acgccagtac accttatcgg
    11221 cccaagcctt gtcccaaggc agcgttttgt tcttggaaac gctgccctac acgttcgcta
    11281 tgcttcaaga acttttctga gcacttcatg atgcatgttt gttccttatt ggttagcttt
    11341 gatgttgtga agtcattgac acagtctgtg aaacatcttt ctaccagatt agagtacaaa
    11401 cgcatgaaat ccttcatttg cttttgttcc actacttttt ggaactcttg ttgttctttc
    11461 ctgcaggtag tcgtgcaatg tatgacttta agatttgtga gcaggaagaa aagggagaat
    11521 cttctaacga taaacccttg aaaaactggg tagactacgc tatgttgagt tgctacgcag
    11581 gctgcacaat tacacgagaa tgctcccgcc taggatttaa ggctaaggga cgtgcaatgc
    11641 agacgacaga tctaaatgac cgtgtcggtg aagtgttcgc caaacttttc ggttaacaca
    11701 tgcagtgatg cacgcgcgat ggtgctaagt tacatatata tatatatgta tagcc
//
