LOCUS       sequence-329112-       11755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constituive spCas9 and tet-inducible GAL2 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_GAL2
FEATURES             Location/Qualifiers
     source          1..11755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1016..1639
                     /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(1089..1108)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     2136..2153
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2270..2291
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2306..2311,2312..2329,2330..2336)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2344..2360
                     /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     2349..2371
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2368..2384
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2368..2384
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2403..2423
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2405..2423
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2441..2839
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     2843..2859
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2843..2859
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2871..6974
                     /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             2871..6974
                     /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             2871..6974
                     /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             6987..7007
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7023..7270
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7039..7057)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(7288..7307)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7289..7307)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7317..7334)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7317..7333)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7326..7348)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7474..7929
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7561..7580)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7771..7792
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(8032..8706)
                     /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(8707..8988)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(9009..9028)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9128..9150
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9188..9206)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9274..9378
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9379..9447,9448..10239)
                     /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(9597..9616)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10410..10998
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10899..10918
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    11382..11400
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        11486..11561
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      11574..11636
                     /label=RPR1 terminator
                     /note="transcription terminator for the S. cerevisiae RNase
                     P RNA gene"
ORIGIN
        1 ccagtacacc ttatcggccc aagccttgtc ccaaggcagc gttttgttct tggaaacgct
       61 gccctacacg ttcgctatgc ttcaagaact tttctgagca cttcatgatg catgtttgtt
      121 ccttattggt tagctttgat gttgtgaagt cattgacaca gtctgtgaaa catctttcta
      181 ccagattaga gtacaaacgc atgaaatcct tcatttgctt ttgttccact actttttgga
      241 actcttgttg ttctttcctg caggtagtcg tgcaatgtat gactttaaga tttgtgagca
      301 ggaagaaaag ggagaatctt ctaacgataa acccttgaaa aactgggtag actacgctat
      361 gttgagttgc tacgcaggct gcacaattac acgagaatgc tcccgcctag gatttaaggc
      421 taagggacgt gcaatgcaga cgacagatct aaatgaccgt gtcggtgaag tgttcgccaa
      481 acttttcggt taacacatgc agtgatgcac gcgcgatggt gctaagttac atatatatat
      541 atatgtatag ccatagtgat gtctaagtaa cctttatggt atatttctta atgtggaaag
      601 atactagcgc gcgcacccac acacaagctt cgtcttttct tgaagaaaag aggaagctcg
      661 ctaaatggga ttccactttc cgttccctgc cagctgatgg aaaaaggtta gtggaacgat
      721 gaagaataaa aagagagatc cactgaggtg aaatttcagc tgacagcgag tttcatgatc
      781 gtgatgaaca atggtaacga gttgtggctg ttgccaggga gggtggttct caacttttaa
      841 tgtatggcca aatcgctact tgggtttgtt atataacaaa gaagaaataa tgaactgatt
      901 ctcttcctcc ttcttgtcct ttcttaattc tgttgtaatt accttccttt gtaatttttt
      961 ttgtaattat tcttcttaat aatccaaaca aacacacata ttacaatata ttaaaatgtc
     1021 tagattagat aaaagtaaag tgattaacag cgcattagag ctgcttaatg aggtcggaat
     1081 cgaaggttta acaacccgta aactcgccca gaagctaggt gtagagcagc ctacattgta
     1141 ttggcatgta aaaaataagc gggctttgct cgacgcctta gccattgaga tgttagatag
     1201 gcaccatact cacttttgcc ctttagaagg ggaaagctgg caagattttt tacgtaataa
     1261 cgctaaaagt tttagatgtg ctttactaag tcatcgcgat ggagcaaaag tacatttagg
     1321 tacacggcct acagaaaaac agtatgaaac tctcgaaaat caattagcct ttttatgcca
     1381 acaaggtttt tcactagaga atgcattata tgcactcagc gctgtggggc attttacttt
     1441 aggttgcgta ttggaagatc aagagcatca agtcgctaaa gaagaaaggg aaacacctac
     1501 tactgatagt atgccgccat tattacgaca agctatcgaa ttatttgatc accaaggtgc
     1561 agagccagcc ttcttattcg gccttgaatt gatcatatgc ggattagaaa aacaacttaa
     1621 atgtgaaagt gggtcttaag tctgaagaat gaatgatttg atgatttctt tttccctcca
     1681 tttttcttac tgaatatatc aatgatatag acttgtatag tttattattt caaattaagt
     1741 agctatatat agtcaagata acgtttgttt gacacgatta cattattcgt cgacatcttt
     1801 tttcagcctg tcgtggtagc aatttgagga gtattattaa ttgaataggt tcattttgcg
     1861 ctcgcataaa cagttttcgt cagggacagt atgttggaat gagtggtaat taatggtgac
     1921 atgacatgtt atagcaataa ccttgatgtt tacatcgtag tttaatgtac accccgcgaa
     1981 ttcgttcaag taggagtgca ccaattgcaa agggaaaagc tgaatgggca gttcgaaccg
     2041 cggtctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     2101 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     2161 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     2221 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     2281 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     2341 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     2401 gcgcaattaa ccctcactaa agggaacaaa agctggagct catagcttca aaatgtttct
     2461 actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac cacttcaaaa
     2521 cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc gttaattacc
     2581 cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa
     2641 aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt gatttttttc
     2701 tctttcgatg acctcccatt gatatttaag ttaataaacg gtcttcaatt tctcaagttt
     2761 cagtttcatt tttcttgttc tattacaact ttttttactt cttgctcatt agaaagaaag
     2821 catagcaatc taatctaagt tttctagaac tagtggatcc cccgggaaaa atggacaaga
     2881 agtactccat tgggctcgat atcggcacaa acagcgtcgg ttgggccgtc attacggacg
     2941 agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa taccgatcgc cacagcataa
     3001 agaagaacct cattggcgcc ctcctgttcg actccgggga gacggccgaa gccacgcggc
     3061 tcaaaagaac agcacggcgc agatataccc gcagaaagaa tcggatctgc tacctgcagg
     3121 agatctttag taatgagatg gctaaggtgg atgactcttt cttccatagg ctggaggagt
     3181 cctttttggt ggaggaggat aaaaagcacg agcgccaccc aatctttggc aatatcgtgg
     3241 acgaggtggc gtaccatgaa aagtacccaa ccatatatca tctgaggaag aagcttgtag
     3301 acagtactga taaggctgac ttgcggttga tctatctcgc gctggcgcat atgatcaaat
     3361 ttcggggaca cttcctcatc gagggggacc tgaacccaga caacagcgat gtcgacaaac
     3421 tctttatcca actggttcag acttacaatc agcttttcga agagaacccg atcaacgcat
     3481 ccggagttga cgccaaagca atcctgagcg ctaggctgtc caaatcccgg cggctcgaaa
     3541 acctcatcgc acagctccct ggggagaaga agaacggcct gtttggtaat cttatcgccc
     3601 tgtcactcgg gctgaccccc aactttaaat ctaacttcga cctggccgaa gatgccaagc
     3661 ttcaactgag caaagacacc tacgatgatg atctcgacaa tctgctggcc cagatcggcg
     3721 accagtacgc agaccttttt ttggcggcaa agaacctgtc agacgccatt ctgctgagtg
     3781 atattctgcg agtgaacacg gagatcacca aagctccgct gagcgctagt atgatcaagc
     3841 gctatgatga gcaccaccaa gacttgactt tgctgaaggc ccttgtcaga cagcaactgc
     3901 ctgagaagta caaggaaatt ttcttcgatc agtctaaaaa tggctacgcc ggatacattg
     3961 acggcggagc aagccaggag gaattttaca aatttattaa gcccatcttg gaaaaaatgg
     4021 acggcaccga ggagctgctg gtaaagctta acagagaaga tctgttgcgc aaacagcgca
     4081 ctttcgacaa tggaagcatc ccccaccaga ttcacctggg cgaactgcac gctatcctca
     4141 ggcggcaaga ggatttctac ccctttttga aagataacag ggaaaagatt gagaaaatcc
     4201 tcacatttcg gataccctac tatgtaggcc ccctcgcccg gggaaattcc agattcgcgt
     4261 ggatgactcg caaatcagaa gagaccatca ctccctggaa cttcgaggaa gtcgtggata
     4321 agggggcctc tgcccagtcc ttcatcgaaa ggatgactaa ctttgataaa aatctgccta
     4381 acgaaaaggt gcttcctaaa cactctctgc tgtacgagta cttcacagtt tataacgagc
     4441 tcaccaaggt caaatacgtc acagaaggga tgagaaagcc agcattcctg tctggagagc
     4501 agaagaaagc tatcgtggac ctcctcttca agacgaaccg gaaagttacc gtgaaacagc
     4561 tcaaagaaga ctatttcaaa aagattgaat gtttcgactc tgttgaaatc agcggagtgg
     4621 aggatcgctt caacgcatcc ctgggaacgt atcacgatct cctgaaaatc attaaagaca
     4681 aggacttcct ggacaatgag gagaacgagg acattcttga ggacattgtc ctcaccctta
     4741 cgttgtttga agatagggag atgattgaag aacgcttgaa aacttacgct catctcttcg
     4801 acgacaaagt catgaaacag ctcaagaggc gccgatatac aggatggggg cggctgtcaa
     4861 gaaaactgat caatgggatc cgagacaagc agagtggaaa gacaatcctg gattttctta
     4921 agtccgatgg atttgccaac cggaacttca tgcagttgat ccatgatgac tctctcacct
     4981 ttaaggagga catccagaaa gcacaagttt ctggccaggg ggacagtctt cacgagcaca
     5041 tcgctaatct tgcaggtagc ccagctatca aaaagggaat actgcagacc gttaaggtcg
     5101 tggatgaact cgtcaaagta atgggaaggc ataagcccga gaatatcgtt atcgagatgg
     5161 cccgagagaa ccaaactacc cagaagggac agaagaacag tagggaaagg atgaagagga
     5221 ttgaagaggg tataaaagaa ctggggtccc aaatccttaa ggaacaccca gttgaaaaca
     5281 cccagcttca gaatgagaag ctctacctgt actacctgca gaacggcagg gacatgtacg
     5341 tggatcagga actggacatc aatcggctct ccgactacga cgtggatcat atcgtgcccc
     5401 agtcttttct caaagatgat tctattgata ataaagtgtt gacaagatcc gataaaaata
     5461 gagggaagag tgataacgtc ccctcagaag aagttgtcaa gaaaatgaaa aattattggc
     5521 ggcagctgct gaacgccaaa ctgatcacac aacggaagtt cgataatctg actaaggctg
     5581 aacgaggtgg cctgtctgag ttggataaag ccggcttcat caaaaggcag cttgttgaga
     5641 cacgccagat caccaagcac gtggcccaaa ttctcgattc acgcatgaac accaagtacg
     5701 atgaaaatga caaactgatt cgagaggtga aagttattac tctgaagtct aagctggtct
     5761 cagatttcag aaaggacttt cagttttata aggtgagaga gatcaacaat taccaccatg
     5821 cgcatgatgc ctacctgaat gcagtggtag gcactgcact tatcaaaaaa tatcccaagc
     5881 ttgaatctga atttgtttac ggagactata aagtgtacga tgttaggaaa atgatcgcaa
     5941 agtctgagca ggaaataggc aaggccaccg ctaagtactt cttttacagc aatattatga
     6001 attttttcaa gaccgagatt acactggcca atggagagat tcggaagcga ccacttatcg
     6061 aaacaaacgg agaaacagga gaaatcgtgt gggacaaggg tagggatttc gcgacagtcc
     6121 ggaaggtcct gtccatgccg caggtgaaca tcgttaaaaa gaccgaagta cagaccggag
     6181 gcttctccaa ggaaagtatc ctcccgaaaa ggaacagcga caagctgatc gcacgcaaaa
     6241 aagattggga ccccaagaaa tacggcggat tcgattctcc tacagtcgct tacagtgtac
     6301 tggttgtggc caaagtggag aaagggaagt ctaaaaaact caaaagcgtc aaggaactgc
     6361 tgggcatcac aatcatggag cgatcaagct tcgaaaaaaa ccccatcgac tttctcgagg
     6421 cgaaaggata taaagaggtc aaaaaagacc tcatcattaa gcttcccaag tactctctct
     6481 ttgagcttga aaacggccgg aaacgaatgc tcgctagtgc gggcgagctg cagaaaggta
     6541 acgagctggc actgccctct aaatacgtta atttcttgta tctggccagc cactatgaaa
     6601 agctcaaagg gtctcccgaa gataatgagc agaagcagct gttcgtggaa caacacaaac
     6661 actaccttga tgagatcatc gagcaaataa gcgaattctc caaaagagtg atcctcgccg
     6721 acgctaacct cgataaggtg ctttctgctt acaataagca cagggataag cccatcaggg
     6781 agcaggcaga aaacattatc cacttgttta ctctgaccaa cttgggcgcg cctgcagcct
     6841 tcaagtactt cgacaccacc atagacagaa agcggtacac ctctacaaag gaggtcctgg
     6901 acgccacact gattcatcag tcaattacgg ggctctatga aacaagaatc gacctctctc
     6961 agctcggtgg agacagcagg gctgacccca agaagaagag gaaggtgtga tctcttctcg
     7021 agtcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac
     7081 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat
     7141 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt
     7201 gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc
     7261 tttaatttgc ggccggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     7321 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     7381 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     7441 tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg
     7501 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     7561 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     7621 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     7681 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     7741 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     7801 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     7861 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     7921 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     7981 agatcggcaa gtgcacaaac aatacttaaa taaatactac tcagtaataa cctatttctt
     8041 agcatttttg acgaaatttg ctattttgtt agagtctttt acaccatttg tctccacacc
     8101 tccgcttaca tcaacaccaa taacgccatt taatctaagc gcatcaccaa cattttctgg
     8161 cgtcagtcca ccagctaaca taaaatgtaa gctttcgggg ctctcttgcc ttccaaccca
     8221 gtcagaaatc gagttccaat ccaaaagttc acctgtccca cctgcttctg aatcaaacaa
     8281 gggaataaac gaatgaggtt tctgtgaagc tgcactgagt agtatgttgc agtcttttgg
     8341 aaatacgagt cttttaataa ctggcaaacc gaggaactct tggtattctt gccacgactc
     8401 atctccatgc agttggacga tatcaatgcc gtaatcattg accagagcca aaacatcctc
     8461 cttaggttga ttacgaaaca cgccaaccaa gtatttcgga gtgcctgaac tatttttata
     8521 tgcttttaca agacttgaaa ttttccttgc aataaccggg tcaattgttc tctttctatt
     8581 gggcacacat ataataccca gcaagtcagc atcggaatct agagcacatt ctgcggcctc
     8641 tgtgctctgc aagccgcaaa ctttcaccaa tggaccagaa ctacctgtga aattaataac
     8701 agacatactc caagctgcct ttgtgtgctt aatcacgtat actcacgtgc tcaatagtca
     8761 ccaatgccct ccctcttggc cctctccttt tcttttttcg accgaattaa ttcttaatcg
     8821 gcaaaaaaag aaaagctccg gatcaagatt gtacgtaagg tgacaagcta tttttcaata
     8881 aagaatatct tccactactg ccatctggcg tcataactgc aaagtacaca tatattacga
     8941 tgctgttcta ttaaatgctt cctatattat atatatagta atgtcgttga tctatggtgc
     9001 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     9061 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     9121 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     9181 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     9241 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     9301 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     9361 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     9421 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     9481 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     9541 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     9601 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     9661 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     9721 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     9781 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     9841 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     9901 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     9961 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
    10021 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
    10081 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
    10141 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
    10201 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
    10261 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
    10321 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
    10381 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
    10441 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
    10501 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
    10561 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
    10621 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
    10681 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
    10741 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
    10801 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
    10861 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
    10921 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
    10981 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
    11041 tggccttttg ctcgggggat ctgccaattg aacataacat ggtagttaca tatactagta
    11101 atatggttcg gcacacatta aaagtataaa aactatctga attacgaatt acatatattg
    11161 gtcataaaaa tcaatcaatc atcgtgtgtt ttatatgtct cttatctaag tataagaata
    11221 tccatagtta atattcactt acgctacctt ttaacctgta atcattgtca acaggatatg
    11281 ttaacgaccc acattgataa acgctagtat ttctttttcc tcttcttatt ggccggctgt
    11341 ctctatactc ccctatagtc tgtttctttt cgtttcgatt gtccctatca gtgatagaga
    11401 tggcgcacat ggtacgctgt ggtgctcgcg gctgggaacg aaactctggg agctgcgatt
    11461 ggcaggatgg ctggctttta ccgtggtttt agagctagaa atagcaagtt aaaataaggc
    11521 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttctc gagccatatc
    11581 caacttccaa tttaatcttt cttttttaat tttcacttat ttgcgataca gaaagaaaaa
    11641 agcgatagta actattgaat tttgtttgga tttggttaga ttagatatgg tttctcttta
    11701 tatttacatg ctaaaaatgg gctacaccag agatacataa ttagatatat atacg
//
