LOCUS       sequence-330353-       11755 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 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_GAL7
FEATURES             Location/Qualifiers
     source          1..11755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    306..324
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        410..485
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      498..560
                     /label=RPR1 terminator
                     /note="transcription terminator for the S. cerevisiae RNase
                     P RNA gene"
     CDS             1695..2318
                     /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(1768..1787)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     2815..2832
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2949..2970
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2985..2990,2991..3008,3009..3015)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3023..3039
                     /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     3028..3050
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3047..3063
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3047..3063
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3082..3102
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3084..3102
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3120..3518
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     3522..3538
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3522..3538
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3550..7653
                     /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             3550..7653
                     /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             3550..7653
                     /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             7666..7686
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7702..7949
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7718..7736)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(7967..7986)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7968..7986)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7996..8013)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7996..8012)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8005..8027)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8153..8608
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8240..8259)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8450..8471
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(8711..9385)
                     /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(9386..9667)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(9688..9707)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9807..9829
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9867..9885)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9953..10057
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10058..10126,10127..10918)
                     /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(10276..10295)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11089..11677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11578..11597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acatggtagt tacatatact agtaatatgg ttcggcacac attaaaagta taaaaactat
       61 ctgaattacg aattacatat attggtcata aaaatcaatc aatcatcgtg tgttttatat
      121 gtctcttatc taagtataag aatatccata gttaatattc acttacgcta ccttttaacc
      181 tgtaatcatt gtcaacagga tatgttaacg acccacattg ataaacgcta gtatttcttt
      241 ttcctcttct tattggccgg ctgtctctat actcccctat agtctgtttc ttttcgtttc
      301 gattgtccct atcagtgata gagatggcgc acatggtacg ctgtggtgct cgcggctggg
      361 aacgaaactc tgggagctgc gattggcagg cctcctgttg acctaaccag ttttagagct
      421 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
      481 ggtgcttttt tctcgagcca tatccaactt ccaatttaat ctttcttttt taattttcac
      541 ttatttgcga tacagaaaga aaaaagcgat agtaactatt gaattttgtt tggatttggt
      601 tagattagat atggtttctc tttatattta catgctaaaa atgggctaca ccagagatac
      661 ataattagat atatatacgc cagtacacct tatcggccca agccttgtcc caaggcagcg
      721 ttttgttctt ggaaacgctg ccctacacgt tcgctatgct tcaagaactt ttctgagcac
      781 ttcatgatgc atgtttgttc cttattggtt agctttgatg ttgtgaagtc attgacacag
      841 tctgtgaaac atctttctac cagattagag tacaaacgca tgaaatcctt catttgcttt
      901 tgttccacta ctttttggaa ctcttgttgt tctttcctgc aggtagtcgt gcaatgtatg
      961 actttaagat ttgtgagcag gaagaaaagg gagaatcttc taacgataaa cccttgaaaa
     1021 actgggtaga ctacgctatg ttgagttgct acgcaggctg cacaattaca cgagaatgct
     1081 cccgcctagg atttaaggct aagggacgtg caatgcagac gacagatcta aatgaccgtg
     1141 tcggtgaagt gttcgccaaa cttttcggtt aacacatgca gtgatgcacg cgcgatggtg
     1201 ctaagttaca tatatatata tatgtatagc catagtgatg tctaagtaac ctttatggta
     1261 tatttcttaa tgtggaaaga tactagcgcg cgcacccaca cacaagcttc gtcttttctt
     1321 gaagaaaaga ggaagctcgc taaatgggat tccactttcc gttccctgcc agctgatgga
     1381 aaaaggttag tggaacgatg aagaataaaa agagagatcc actgaggtga aatttcagct
     1441 gacagcgagt ttcatgatcg tgatgaacaa tggtaacgag ttgtggctgt tgccagggag
     1501 ggtggttctc aacttttaat gtatggccaa atcgctactt gggtttgtta tataacaaag
     1561 aagaaataat gaactgattc tcttcctcct tcttgtcctt tcttaattct gttgtaatta
     1621 ccttcctttg taattttttt tgtaattatt cttcttaata atccaaacaa acacacatat
     1681 tacaatatat taaaatgtct agattagata aaagtaaagt gattaacagc gcattagagc
     1741 tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa actcgcccag aagctaggtg
     1801 tagagcagcc tacattgtat tggcatgtaa aaaataagcg ggctttgctc gacgccttag
     1861 ccattgagat gttagatagg caccatactc acttttgccc tttagaaggg gaaagctggc
     1921 aagatttttt acgtaataac gctaaaagtt ttagatgtgc tttactaagt catcgcgatg
     1981 gagcaaaagt acatttaggt acacggccta cagaaaaaca gtatgaaact ctcgaaaatc
     2041 aattagcctt tttatgccaa caaggttttt cactagagaa tgcattatat gcactcagcg
     2101 ctgtggggca ttttacttta ggttgcgtat tggaagatca agagcatcaa gtcgctaaag
     2161 aagaaaggga aacacctact actgatagta tgccgccatt attacgacaa gctatcgaat
     2221 tatttgatca ccaaggtgca gagccagcct tcttattcgg ccttgaattg atcatatgcg
     2281 gattagaaaa acaacttaaa tgtgaaagtg ggtcttaagt ctgaagaatg aatgatttga
     2341 tgatttcttt ttccctccat ttttcttact gaatatatca atgatataga cttgtatagt
     2401 ttattatttc aaattaagta gctatatata gtcaagataa cgtttgtttg acacgattac
     2461 attattcgtc gacatctttt ttcagcctgt cgtggtagca atttgaggag tattattaat
     2521 tgaataggtt cattttgcgc tcgcataaac agttttcgtc agggacagta tgttggaatg
     2581 agtggtaatt aatggtgaca tgacatgtta tagcaataac cttgatgttt acatcgtagt
     2641 ttaatgtaca ccccgcgaat tcgttcaagt aggagtgcac caattgcaaa gggaaaagct
     2701 gaatgggcag ttcgaaccgc ggtctttcct gcgttatccc ctgattctgt ggataaccgt
     2761 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     2821 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     2881 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     2941 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     3001 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     3061 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctc
     3121 atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca
     3181 tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc
     3241 tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt
     3301 tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat
     3361 tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt taataaacgg
     3421 tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc
     3481 ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttctagaact agtggatccc
     3541 ccgggaaaaa tggacaagaa gtactccatt gggctcgata tcggcacaaa cagcgtcggt
     3601 tgggccgtca ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt tctgggcaat
     3661 accgatcgcc acagcataaa gaagaacctc attggcgccc tcctgttcga ctccggggag
     3721 acggccgaag ccacgcggct caaaagaaca gcacggcgca gatatacccg cagaaagaat
     3781 cggatctgct acctgcagga gatctttagt aatgagatgg ctaaggtgga tgactctttc
     3841 ttccataggc tggaggagtc ctttttggtg gaggaggata aaaagcacga gcgccaccca
     3901 atctttggca atatcgtgga cgaggtggcg taccatgaaa agtacccaac catatatcat
     3961 ctgaggaaga agcttgtaga cagtactgat aaggctgact tgcggttgat ctatctcgcg
     4021 ctggcgcata tgatcaaatt tcggggacac ttcctcatcg agggggacct gaacccagac
     4081 aacagcgatg tcgacaaact ctttatccaa ctggttcaga cttacaatca gcttttcgaa
     4141 gagaacccga tcaacgcatc cggagttgac gccaaagcaa tcctgagcgc taggctgtcc
     4201 aaatcccggc ggctcgaaaa cctcatcgca cagctccctg gggagaagaa gaacggcctg
     4261 tttggtaatc ttatcgccct gtcactcggg ctgaccccca actttaaatc taacttcgac
     4321 ctggccgaag atgccaagct tcaactgagc aaagacacct acgatgatga tctcgacaat
     4381 ctgctggccc agatcggcga ccagtacgca gacctttttt tggcggcaaa gaacctgtca
     4441 gacgccattc tgctgagtga tattctgcga gtgaacacgg agatcaccaa agctccgctg
     4501 agcgctagta tgatcaagcg ctatgatgag caccaccaag acttgacttt gctgaaggcc
     4561 cttgtcagac agcaactgcc tgagaagtac aaggaaattt tcttcgatca gtctaaaaat
     4621 ggctacgccg gatacattga cggcggagca agccaggagg aattttacaa atttattaag
     4681 cccatcttgg aaaaaatgga cggcaccgag gagctgctgg taaagcttaa cagagaagat
     4741 ctgttgcgca aacagcgcac tttcgacaat ggaagcatcc cccaccagat tcacctgggc
     4801 gaactgcacg ctatcctcag gcggcaagag gatttctacc cctttttgaa agataacagg
     4861 gaaaagattg agaaaatcct cacatttcgg ataccctact atgtaggccc cctcgcccgg
     4921 ggaaattcca gattcgcgtg gatgactcgc aaatcagaag agaccatcac tccctggaac
     4981 ttcgaggaag tcgtggataa gggggcctct gcccagtcct tcatcgaaag gatgactaac
     5041 tttgataaaa atctgcctaa cgaaaaggtg cttcctaaac actctctgct gtacgagtac
     5101 ttcacagttt ataacgagct caccaaggtc aaatacgtca cagaagggat gagaaagcca
     5161 gcattcctgt ctggagagca gaagaaagct atcgtggacc tcctcttcaa gacgaaccgg
     5221 aaagttaccg tgaaacagct caaagaagac tatttcaaaa agattgaatg tttcgactct
     5281 gttgaaatca gcggagtgga ggatcgcttc aacgcatccc tgggaacgta tcacgatctc
     5341 ctgaaaatca ttaaagacaa ggacttcctg gacaatgagg agaacgagga cattcttgag
     5401 gacattgtcc tcacccttac gttgtttgaa gatagggaga tgattgaaga acgcttgaaa
     5461 acttacgctc atctcttcga cgacaaagtc atgaaacagc tcaagaggcg ccgatataca
     5521 ggatgggggc ggctgtcaag aaaactgatc aatgggatcc gagacaagca gagtggaaag
     5581 acaatcctgg attttcttaa gtccgatgga tttgccaacc ggaacttcat gcagttgatc
     5641 catgatgact ctctcacctt taaggaggac atccagaaag cacaagtttc tggccagggg
     5701 gacagtcttc acgagcacat cgctaatctt gcaggtagcc cagctatcaa aaagggaata
     5761 ctgcagaccg ttaaggtcgt ggatgaactc gtcaaagtaa tgggaaggca taagcccgag
     5821 aatatcgtta tcgagatggc ccgagagaac caaactaccc agaagggaca gaagaacagt
     5881 agggaaagga tgaagaggat tgaagagggt ataaaagaac tggggtccca aatccttaag
     5941 gaacacccag ttgaaaacac ccagcttcag aatgagaagc tctacctgta ctacctgcag
     6001 aacggcaggg acatgtacgt ggatcaggaa ctggacatca atcggctctc cgactacgac
     6061 gtggatcata tcgtgcccca gtcttttctc aaagatgatt ctattgataa taaagtgttg
     6121 acaagatccg ataaaaatag agggaagagt gataacgtcc cctcagaaga agttgtcaag
     6181 aaaatgaaaa attattggcg gcagctgctg aacgccaaac tgatcacaca acggaagttc
     6241 gataatctga ctaaggctga acgaggtggc ctgtctgagt tggataaagc cggcttcatc
     6301 aaaaggcagc ttgttgagac acgccagatc accaagcacg tggcccaaat tctcgattca
     6361 cgcatgaaca ccaagtacga tgaaaatgac aaactgattc gagaggtgaa agttattact
     6421 ctgaagtcta agctggtctc agatttcaga aaggactttc agttttataa ggtgagagag
     6481 atcaacaatt accaccatgc gcatgatgcc tacctgaatg cagtggtagg cactgcactt
     6541 atcaaaaaat atcccaagct tgaatctgaa tttgtttacg gagactataa agtgtacgat
     6601 gttaggaaaa tgatcgcaaa gtctgagcag gaaataggca aggccaccgc taagtacttc
     6661 ttttacagca atattatgaa ttttttcaag accgagatta cactggccaa tggagagatt
     6721 cggaagcgac cacttatcga aacaaacgga gaaacaggag aaatcgtgtg ggacaagggt
     6781 agggatttcg cgacagtccg gaaggtcctg tccatgccgc aggtgaacat cgttaaaaag
     6841 accgaagtac agaccggagg cttctccaag gaaagtatcc tcccgaaaag gaacagcgac
     6901 aagctgatcg cacgcaaaaa agattgggac cccaagaaat acggcggatt cgattctcct
     6961 acagtcgctt acagtgtact ggttgtggcc aaagtggaga aagggaagtc taaaaaactc
     7021 aaaagcgtca aggaactgct gggcatcaca atcatggagc gatcaagctt cgaaaaaaac
     7081 cccatcgact ttctcgaggc gaaaggatat aaagaggtca aaaaagacct catcattaag
     7141 cttcccaagt actctctctt tgagcttgaa aacggccgga aacgaatgct cgctagtgcg
     7201 ggcgagctgc agaaaggtaa cgagctggca ctgccctcta aatacgttaa tttcttgtat
     7261 ctggccagcc actatgaaaa gctcaaaggg tctcccgaag ataatgagca gaagcagctg
     7321 ttcgtggaac aacacaaaca ctaccttgat gagatcatcg agcaaataag cgaattctcc
     7381 aaaagagtga tcctcgccga cgctaacctc gataaggtgc tttctgctta caataagcac
     7441 agggataagc ccatcaggga gcaggcagaa aacattatcc acttgtttac tctgaccaac
     7501 ttgggcgcgc ctgcagcctt caagtacttc gacaccacca tagacagaaa gcggtacacc
     7561 tctacaaagg aggtcctgga cgccacactg attcatcagt caattacggg gctctatgaa
     7621 acaagaatcg acctctctca gctcggtgga gacagcaggg ctgaccccaa gaagaagagg
     7681 aaggtgtgat ctcttctcga gtcatgtaat tagttatgtc acgcttacat tcacgccctc
     7741 cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta
     7801 tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
     7861 tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg
     7921 ttttgggacg ctcgaaggct ttaatttgcg gccggtaccc aattcgccct atagtgagtc
     7981 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     8041 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     8101 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     8161 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     8221 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     8281 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     8341 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     8401 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     8461 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     8521 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     8581 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     8641 gcggtatttc acaccgcata gatcggcaag tgcacaaaca atacttaaat aaatactact
     8701 cagtaataac ctatttctta gcatttttga cgaaatttgc tattttgtta gagtctttta
     8761 caccatttgt ctccacacct ccgcttacat caacaccaat aacgccattt aatctaagcg
     8821 catcaccaac attttctggc gtcagtccac cagctaacat aaaatgtaag ctttcggggc
     8881 tctcttgcct tccaacccag tcagaaatcg agttccaatc caaaagttca cctgtcccac
     8941 ctgcttctga atcaaacaag ggaataaacg aatgaggttt ctgtgaagct gcactgagta
     9001 gtatgttgca gtcttttgga aatacgagtc ttttaataac tggcaaaccg aggaactctt
     9061 ggtattcttg ccacgactca tctccatgca gttggacgat atcaatgccg taatcattga
     9121 ccagagccaa aacatcctcc ttaggttgat tacgaaacac gccaaccaag tatttcggag
     9181 tgcctgaact atttttatat gcttttacaa gacttgaaat tttccttgca ataaccgggt
     9241 caattgttct ctttctattg ggcacacata taatacccag caagtcagca tcggaatcta
     9301 gagcacattc tgcggcctct gtgctctgca agccgcaaac tttcaccaat ggaccagaac
     9361 tacctgtgaa attaataaca gacatactcc aagctgcctt tgtgtgctta atcacgtata
     9421 ctcacgtgct caatagtcac caatgccctc cctcttggcc ctctcctttt cttttttcga
     9481 ccgaattaat tcttaatcgg caaaaaaaga aaagctccgg atcaagattg tacgtaaggt
     9541 gacaagctat ttttcaataa agaatatctt ccactactgc catctggcgt cataactgca
     9601 aagtacacat atattacgat gctgttctat taaatgcttc ctatattata tatatagtaa
     9661 tgtcgttgat ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     9721 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     9781 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     9841 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     9901 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     9961 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
    10021 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
    10081 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
    10141 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
    10201 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
    10261 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
    10321 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
    10381 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
    10441 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
    10501 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
    10561 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
    10621 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
    10681 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
    10741 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
    10801 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
    10861 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
    10921 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
    10981 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
    11041 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
    11101 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
    11161 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
    11221 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
    11281 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
    11341 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
    11401 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
    11461 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
    11521 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
    11581 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
    11641 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
    11701 acggttcctg gccttttgct ggccttttgc tcgggggatc tgccaattga acata
//
