LOCUS       sequence-329131-       11755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constituive spCas9 expression and tet-inducible 
            expression of sgRNA binding to the promoter of YNR071C 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_YNR071C
FEATURES             Location/Qualifiers
     source          1..11755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      13..75
                     /label=RPR1 terminator
                     /note="transcription terminator for the S. cerevisiae RNase
                     P RNA gene"
     CDS             1210..1833
                     /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(1283..1302)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     2330..2347
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2464..2485
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2500..2505,2506..2523,2524..2530)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2538..2554
                     /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     2543..2565
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2562..2578
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2562..2578
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2597..2617
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2599..2617
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2635..3033
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     3037..3053
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3037..3053
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3065..7168
                     /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             3065..7168
                     /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             3065..7168
                     /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             7181..7201
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7217..7464
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7233..7251)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(7482..7501)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7483..7501)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7511..7528)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7511..7527)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7520..7542)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7668..8123
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7755..7774)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7965..7986
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(8226..8900)
                     /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(8901..9182)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(9203..9222)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9322..9344
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9382..9400)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9468..9572
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9573..9641,9642..10433)
                     /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(9791..9810)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10604..11192
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11093..11112
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    11576..11594
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        11680..11755
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 ttttttctcg agccatatcc aacttccaat ttaatctttc ttttttaatt ttcacttatt
       61 tgcgatacag aaagaaaaaa gcgatagtaa ctattgaatt ttgtttggat ttggttagat
      121 tagatatggt ttctctttat atttacatgc taaaaatggg ctacaccaga gatacataat
      181 tagatatata tacgccagta caccttatcg gcccaagcct tgtcccaagg cagcgttttg
      241 ttcttggaaa cgctgcccta cacgttcgct atgcttcaag aacttttctg agcacttcat
      301 gatgcatgtt tgttccttat tggttagctt tgatgttgtg aagtcattga cacagtctgt
      361 gaaacatctt tctaccagat tagagtacaa acgcatgaaa tccttcattt gcttttgttc
      421 cactactttt tggaactctt gttgttcttt cctgcaggta gtcgtgcaat gtatgacttt
      481 aagatttgtg agcaggaaga aaagggagaa tcttctaacg ataaaccctt gaaaaactgg
      541 gtagactacg ctatgttgag ttgctacgca ggctgcacaa ttacacgaga atgctcccgc
      601 ctaggattta aggctaaggg acgtgcaatg cagacgacag atctaaatga ccgtgtcggt
      661 gaagtgttcg ccaaactttt cggttaacac atgcagtgat gcacgcgcga tggtgctaag
      721 ttacatatat atatatatgt atagccatag tgatgtctaa gtaaccttta tggtatattt
      781 cttaatgtgg aaagatacta gcgcgcgcac ccacacacaa gcttcgtctt ttcttgaaga
      841 aaagaggaag ctcgctaaat gggattccac tttccgttcc ctgccagctg atggaaaaag
      901 gttagtggaa cgatgaagaa taaaaagaga gatccactga ggtgaaattt cagctgacag
      961 cgagtttcat gatcgtgatg aacaatggta acgagttgtg gctgttgcca gggagggtgg
     1021 ttctcaactt ttaatgtatg gccaaatcgc tacttgggtt tgttatataa caaagaagaa
     1081 ataatgaact gattctcttc ctccttcttg tcctttctta attctgttgt aattaccttc
     1141 ctttgtaatt ttttttgtaa ttattcttct taataatcca aacaaacaca catattacaa
     1201 tatattaaaa tgtctagatt agataaaagt aaagtgatta acagcgcatt agagctgctt
     1261 aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg cccagaagct aggtgtagag
     1321 cagcctacat tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt
     1381 gagatgttag ataggcacca tactcacttt tgccctttag aaggggaaag ctggcaagat
     1441 tttttacgta ataacgctaa aagttttaga tgtgctttac taagtcatcg cgatggagca
     1501 aaagtacatt taggtacacg gcctacagaa aaacagtatg aaactctcga aaatcaatta
     1561 gcctttttat gccaacaagg tttttcacta gagaatgcat tatatgcact cagcgctgtg
     1621 gggcatttta ctttaggttg cgtattggaa gatcaagagc atcaagtcgc taaagaagaa
     1681 agggaaacac ctactactga tagtatgccg ccattattac gacaagctat cgaattattt
     1741 gatcaccaag gtgcagagcc agccttctta ttcggccttg aattgatcat atgcggatta
     1801 gaaaaacaac ttaaatgtga aagtgggtct taagtctgaa gaatgaatga tttgatgatt
     1861 tctttttccc tccatttttc ttactgaata tatcaatgat atagacttgt atagtttatt
     1921 atttcaaatt aagtagctat atatagtcaa gataacgttt gtttgacacg attacattat
     1981 tcgtcgacat cttttttcag cctgtcgtgg tagcaatttg aggagtatta ttaattgaat
     2041 aggttcattt tgcgctcgca taaacagttt tcgtcaggga cagtatgttg gaatgagtgg
     2101 taattaatgg tgacatgaca tgttatagca ataaccttga tgtttacatc gtagtttaat
     2161 gtacaccccg cgaattcgtt caagtaggag tgcaccaatt gcaaagggaa aagctgaatg
     2221 ggcagttcga accgcggtct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2281 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     2341 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     2401 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     2461 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     2521 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     2581 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctcatagc
     2641 ttcaaaatgt ttctactcct tttttactct tccagatttt ctcggactcc gcgcatcgcc
     2701 gtaccacttc aaaacaccca agcacagcat actaaatttc ccctctttct tcctctaggg
     2761 tgtcgttaat tacccgtact aaaggtttgg aaaagaaaaa agagaccgcc tcgtttcttt
     2821 ttcttcgtcg aaaaaggcaa taaaaatttt tatcacgttt ctttttcttg aaaatttttt
     2881 ttttgatttt tttctctttc gatgacctcc cattgatatt taagttaata aacggtcttc
     2941 aatttctcaa gtttcagttt catttttctt gttctattac aacttttttt acttcttgct
     3001 cattagaaag aaagcatagc aatctaatct aagttttcta gaactagtgg atcccccggg
     3061 aaaaatggac aagaagtact ccattgggct cgatatcggc acaaacagcg tcggttgggc
     3121 cgtcattacg gacgagtaca aggtgccgag caaaaaattc aaagttctgg gcaataccga
     3181 tcgccacagc ataaagaaga acctcattgg cgccctcctg ttcgactccg gggagacggc
     3241 cgaagccacg cggctcaaaa gaacagcacg gcgcagatat acccgcagaa agaatcggat
     3301 ctgctacctg caggagatct ttagtaatga gatggctaag gtggatgact ctttcttcca
     3361 taggctggag gagtcctttt tggtggagga ggataaaaag cacgagcgcc acccaatctt
     3421 tggcaatatc gtggacgagg tggcgtacca tgaaaagtac ccaaccatat atcatctgag
     3481 gaagaagctt gtagacagta ctgataaggc tgacttgcgg ttgatctatc tcgcgctggc
     3541 gcatatgatc aaatttcggg gacacttcct catcgagggg gacctgaacc cagacaacag
     3601 cgatgtcgac aaactcttta tccaactggt tcagacttac aatcagcttt tcgaagagaa
     3661 cccgatcaac gcatccggag ttgacgccaa agcaatcctg agcgctaggc tgtccaaatc
     3721 ccggcggctc gaaaacctca tcgcacagct ccctggggag aagaagaacg gcctgtttgg
     3781 taatcttatc gccctgtcac tcgggctgac ccccaacttt aaatctaact tcgacctggc
     3841 cgaagatgcc aagcttcaac tgagcaaaga cacctacgat gatgatctcg acaatctgct
     3901 ggcccagatc ggcgaccagt acgcagacct ttttttggcg gcaaagaacc tgtcagacgc
     3961 cattctgctg agtgatattc tgcgagtgaa cacggagatc accaaagctc cgctgagcgc
     4021 tagtatgatc aagcgctatg atgagcacca ccaagacttg actttgctga aggcccttgt
     4081 cagacagcaa ctgcctgaga agtacaagga aattttcttc gatcagtcta aaaatggcta
     4141 cgccggatac attgacggcg gagcaagcca ggaggaattt tacaaattta ttaagcccat
     4201 cttggaaaaa atggacggca ccgaggagct gctggtaaag cttaacagag aagatctgtt
     4261 gcgcaaacag cgcactttcg acaatggaag catcccccac cagattcacc tgggcgaact
     4321 gcacgctatc ctcaggcggc aagaggattt ctaccccttt ttgaaagata acagggaaaa
     4381 gattgagaaa atcctcacat ttcggatacc ctactatgta ggccccctcg cccggggaaa
     4441 ttccagattc gcgtggatga ctcgcaaatc agaagagacc atcactccct ggaacttcga
     4501 ggaagtcgtg gataaggggg cctctgccca gtccttcatc gaaaggatga ctaactttga
     4561 taaaaatctg cctaacgaaa aggtgcttcc taaacactct ctgctgtacg agtacttcac
     4621 agtttataac gagctcacca aggtcaaata cgtcacagaa gggatgagaa agccagcatt
     4681 cctgtctgga gagcagaaga aagctatcgt ggacctcctc ttcaagacga accggaaagt
     4741 taccgtgaaa cagctcaaag aagactattt caaaaagatt gaatgtttcg actctgttga
     4801 aatcagcgga gtggaggatc gcttcaacgc atccctggga acgtatcacg atctcctgaa
     4861 aatcattaaa gacaaggact tcctggacaa tgaggagaac gaggacattc ttgaggacat
     4921 tgtcctcacc cttacgttgt ttgaagatag ggagatgatt gaagaacgct tgaaaactta
     4981 cgctcatctc ttcgacgaca aagtcatgaa acagctcaag aggcgccgat atacaggatg
     5041 ggggcggctg tcaagaaaac tgatcaatgg gatccgagac aagcagagtg gaaagacaat
     5101 cctggatttt cttaagtccg atggatttgc caaccggaac ttcatgcagt tgatccatga
     5161 tgactctctc acctttaagg aggacatcca gaaagcacaa gtttctggcc agggggacag
     5221 tcttcacgag cacatcgcta atcttgcagg tagcccagct atcaaaaagg gaatactgca
     5281 gaccgttaag gtcgtggatg aactcgtcaa agtaatggga aggcataagc ccgagaatat
     5341 cgttatcgag atggcccgag agaaccaaac tacccagaag ggacagaaga acagtaggga
     5401 aaggatgaag aggattgaag agggtataaa agaactgggg tcccaaatcc ttaaggaaca
     5461 cccagttgaa aacacccagc ttcagaatga gaagctctac ctgtactacc tgcagaacgg
     5521 cagggacatg tacgtggatc aggaactgga catcaatcgg ctctccgact acgacgtgga
     5581 tcatatcgtg ccccagtctt ttctcaaaga tgattctatt gataataaag tgttgacaag
     5641 atccgataaa aatagaggga agagtgataa cgtcccctca gaagaagttg tcaagaaaat
     5701 gaaaaattat tggcggcagc tgctgaacgc caaactgatc acacaacgga agttcgataa
     5761 tctgactaag gctgaacgag gtggcctgtc tgagttggat aaagccggct tcatcaaaag
     5821 gcagcttgtt gagacacgcc agatcaccaa gcacgtggcc caaattctcg attcacgcat
     5881 gaacaccaag tacgatgaaa atgacaaact gattcgagag gtgaaagtta ttactctgaa
     5941 gtctaagctg gtctcagatt tcagaaagga ctttcagttt tataaggtga gagagatcaa
     6001 caattaccac catgcgcatg atgcctacct gaatgcagtg gtaggcactg cacttatcaa
     6061 aaaatatccc aagcttgaat ctgaatttgt ttacggagac tataaagtgt acgatgttag
     6121 gaaaatgatc gcaaagtctg agcaggaaat aggcaaggcc accgctaagt acttctttta
     6181 cagcaatatt atgaattttt tcaagaccga gattacactg gccaatggag agattcggaa
     6241 gcgaccactt atcgaaacaa acggagaaac aggagaaatc gtgtgggaca agggtaggga
     6301 tttcgcgaca gtccggaagg tcctgtccat gccgcaggtg aacatcgtta aaaagaccga
     6361 agtacagacc ggaggcttct ccaaggaaag tatcctcccg aaaaggaaca gcgacaagct
     6421 gatcgcacgc aaaaaagatt gggaccccaa gaaatacggc ggattcgatt ctcctacagt
     6481 cgcttacagt gtactggttg tggccaaagt ggagaaaggg aagtctaaaa aactcaaaag
     6541 cgtcaaggaa ctgctgggca tcacaatcat ggagcgatca agcttcgaaa aaaaccccat
     6601 cgactttctc gaggcgaaag gatataaaga ggtcaaaaaa gacctcatca ttaagcttcc
     6661 caagtactct ctctttgagc ttgaaaacgg ccggaaacga atgctcgcta gtgcgggcga
     6721 gctgcagaaa ggtaacgagc tggcactgcc ctctaaatac gttaatttct tgtatctggc
     6781 cagccactat gaaaagctca aagggtctcc cgaagataat gagcagaagc agctgttcgt
     6841 ggaacaacac aaacactacc ttgatgagat catcgagcaa ataagcgaat tctccaaaag
     6901 agtgatcctc gccgacgcta acctcgataa ggtgctttct gcttacaata agcacaggga
     6961 taagcccatc agggagcagg cagaaaacat tatccacttg tttactctga ccaacttggg
     7021 cgcgcctgca gccttcaagt acttcgacac caccatagac agaaagcggt acacctctac
     7081 aaaggaggtc ctggacgcca cactgattca tcagtcaatt acggggctct atgaaacaag
     7141 aatcgacctc tctcagctcg gtggagacag cagggctgac cccaagaaga agaggaaggt
     7201 gtgatctctt ctcgagtcat gtaattagtt atgtcacgct tacattcacg ccctcccccc
     7261 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     7321 ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
     7381 ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg
     7441 ggacgctcga aggctttaat ttgcggccgg tacccaattc gccctatagt gagtcgtatt
     7501 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     7561 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     7621 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag
     7681 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     7741 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     7801 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     7861 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     7921 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     7981 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     8041 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     8101 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     8161 atttcacacc gcatagatcg gcaagtgcac aaacaatact taaataaata ctactcagta
     8221 ataacctatt tcttagcatt tttgacgaaa tttgctattt tgttagagtc ttttacacca
     8281 tttgtctcca cacctccgct tacatcaaca ccaataacgc catttaatct aagcgcatca
     8341 ccaacatttt ctggcgtcag tccaccagct aacataaaat gtaagctttc ggggctctct
     8401 tgccttccaa cccagtcaga aatcgagttc caatccaaaa gttcacctgt cccacctgct
     8461 tctgaatcaa acaagggaat aaacgaatga ggtttctgtg aagctgcact gagtagtatg
     8521 ttgcagtctt ttggaaatac gagtctttta ataactggca aaccgaggaa ctcttggtat
     8581 tcttgccacg actcatctcc atgcagttgg acgatatcaa tgccgtaatc attgaccaga
     8641 gccaaaacat cctccttagg ttgattacga aacacgccaa ccaagtattt cggagtgcct
     8701 gaactatttt tatatgcttt tacaagactt gaaattttcc ttgcaataac cgggtcaatt
     8761 gttctctttc tattgggcac acatataata cccagcaagt cagcatcgga atctagagca
     8821 cattctgcgg cctctgtgct ctgcaagccg caaactttca ccaatggacc agaactacct
     8881 gtgaaattaa taacagacat actccaagct gcctttgtgt gcttaatcac gtatactcac
     8941 gtgctcaata gtcaccaatg ccctccctct tggccctctc cttttctttt ttcgaccgaa
     9001 ttaattctta atcggcaaaa aaagaaaagc tccggatcaa gattgtacgt aaggtgacaa
     9061 gctatttttc aataaagaat atcttccact actgccatct ggcgtcataa ctgcaaagta
     9121 cacatatatt acgatgctgt tctattaaat gcttcctata ttatatatat agtaatgtcg
     9181 ttgatctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     9241 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     9301 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     9361 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     9421 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     9481 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     9541 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     9601 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     9661 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     9721 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     9781 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     9841 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     9901 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     9961 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
    10021 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
    10081 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
    10141 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
    10201 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
    10261 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
    10321 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
    10381 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
    10441 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
    10501 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
    10561 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
    10621 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
    10681 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
    10741 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
    10801 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
    10861 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
    10921 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
    10981 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
    11041 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
    11101 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
    11161 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
    11221 tcctggcctt ttgctggcct tttgctcggg ggatctgcca attgaacata acatggtagt
    11281 tacatatact agtaatatgg ttcggcacac attaaaagta taaaaactat ctgaattacg
    11341 aattacatat attggtcata aaaatcaatc aatcatcgtg tgttttatat gtctcttatc
    11401 taagtataag aatatccata gttaatattc acttacgcta ccttttaacc tgtaatcatt
    11461 gtcaacagga tatgttaacg acccacattg ataaacgcta gtatttcttt ttcctcttct
    11521 tattggccgg ctgtctctat actcccctat agtctgtttc ttttcgtttc gattgtccct
    11581 atcagtgata gagatggcgc acatggtacg ctgtggtgct cgcggctggg aacgaaactc
    11641 tgggagctgc gattggcagt aactgatctt aataacacgg ttttagagct agaaatagca
    11701 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgc
//
