LOCUS       sequence-330349-       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 RPL39 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_RPL39
FEATURES             Location/Qualifiers
     source          1..11755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             742..1365
                     /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(815..834)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     1862..1879
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1996..2017
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2032..2037,2038..2055,2056..2062)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2070..2086
                     /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     2075..2097
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2094..2110
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2094..2110
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2129..2149
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2131..2149
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2167..2565
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     2569..2585
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2569..2585
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2597..6700
                     /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             2597..6700
                     /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             2597..6700
                     /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             6713..6733
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6749..6996
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6765..6783)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(7014..7033)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7015..7033)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7043..7060)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7043..7059)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7052..7074)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7200..7655
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7287..7306)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7497..7518
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7758..8432)
                     /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(8433..8714)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(8735..8754)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8854..8876
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8914..8932)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9000..9104
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9105..9173,9174..9965)
                     /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(9323..9342)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10136..10724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10625..10644
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    11108..11126
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        11212..11287
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      11300..11362
                     /label=RPR1 terminator
                     /note="transcription terminator for the S. cerevisiae RNase
                     P RNA gene"
ORIGIN
        1 atgtatgact ttaagatttg tgagcaggaa gaaaagggag aatcttctaa cgataaaccc
       61 ttgaaaaact gggtagacta cgctatgttg agttgctacg caggctgcac aattacacga
      121 gaatgctccc gcctaggatt taaggctaag ggacgtgcaa tgcagacgac agatctaaat
      181 gaccgtgtcg gtgaagtgtt cgccaaactt ttcggttaac acatgcagtg atgcacgcgc
      241 gatggtgcta agttacatat atatatatat gtatagccat agtgatgtct aagtaacctt
      301 tatggtatat ttcttaatgt ggaaagatac tagcgcgcgc acccacacac aagcttcgtc
      361 ttttcttgaa gaaaagagga agctcgctaa atgggattcc actttccgtt ccctgccagc
      421 tgatggaaaa aggttagtgg aacgatgaag aataaaaaga gagatccact gaggtgaaat
      481 ttcagctgac agcgagtttc atgatcgtga tgaacaatgg taacgagttg tggctgttgc
      541 cagggagggt ggttctcaac ttttaatgta tggccaaatc gctacttggg tttgttatat
      601 aacaaagaag aaataatgaa ctgattctct tcctccttct tgtcctttct taattctgtt
      661 gtaattacct tcctttgtaa ttttttttgt aattattctt cttaataatc caaacaaaca
      721 cacatattac aatatattaa aatgtctaga ttagataaaa gtaaagtgat taacagcgca
      781 ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag
      841 ctaggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac
      901 gccttagcca ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa
      961 agctggcaag attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat
     1021 cgcgatggag caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc
     1081 gaaaatcaat tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca
     1141 ctcagcgctg tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc
     1201 gctaaagaag aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct
     1261 atcgaattat ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc
     1321 atatgcggat tagaaaaaca acttaaatgt gaaagtgggt cttaagtctg aagaatgaat
     1381 gatttgatga tttctttttc cctccatttt tcttactgaa tatatcaatg atatagactt
     1441 gtatagttta ttatttcaaa ttaagtagct atatatagtc aagataacgt ttgtttgaca
     1501 cgattacatt attcgtcgac atcttttttc agcctgtcgt ggtagcaatt tgaggagtat
     1561 tattaattga ataggttcat tttgcgctcg cataaacagt tttcgtcagg gacagtatgt
     1621 tggaatgagt ggtaattaat ggtgacatga catgttatag caataacctt gatgtttaca
     1681 tcgtagttta atgtacaccc cgcgaattcg ttcaagtagg agtgcaccaa ttgcaaaggg
     1741 aaaagctgaa tgggcagttc gaaccgcggt ctttcctgcg ttatcccctg attctgtgga
     1801 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1861 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     1921 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     1981 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     2041 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     2101 cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct
     2161 ggagctcata gcttcaaaat gtttctactc cttttttact cttccagatt ttctcggact
     2221 ccgcgcatcg ccgtaccact tcaaaacacc caagcacagc atactaaatt tcccctcttt
     2281 cttcctctag ggtgtcgtta attacccgta ctaaaggttt ggaaaagaaa aaagagaccg
     2341 cctcgtttct ttttcttcgt cgaaaaaggc aataaaaatt tttatcacgt ttctttttct
     2401 tgaaaatttt ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa
     2461 taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt
     2521 ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagttttc tagaactagt
     2581 ggatcccccg ggaaaaatgg acaagaagta ctccattggg ctcgatatcg gcacaaacag
     2641 cgtcggttgg gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct
     2701 gggcaatacc gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc
     2761 cggggagacg gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag
     2821 aaagaatcgg atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga
     2881 ctctttcttc cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg
     2941 ccacccaatc tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat
     3001 atatcatctg aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta
     3061 tctcgcgctg gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa
     3121 cccagacaac agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct
     3181 tttcgaagag aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag
     3241 gctgtccaaa tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa
     3301 cggcctgttt ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa
     3361 cttcgacctg gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct
     3421 cgacaatctg ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa
     3481 cctgtcagac gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc
     3541 tccgctgagc gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct
     3601 gaaggccctt gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc
     3661 taaaaatggc tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt
     3721 tattaagccc atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag
     3781 agaagatctg ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca
     3841 cctgggcgaa ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga
     3901 taacagggaa aagattgaga aaatcctcac atttcggata ccctactatg taggccccct
     3961 cgcccgggga aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc
     4021 ctggaacttc gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat
     4081 gactaacttt gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta
     4141 cgagtacttc acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag
     4201 aaagccagca ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac
     4261 gaaccggaaa gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt
     4321 cgactctgtt gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca
     4381 cgatctcctg aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat
     4441 tcttgaggac attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg
     4501 cttgaaaact tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg
     4561 atatacagga tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag
     4621 tggaaagaca atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca
     4681 gttgatccat gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg
     4741 ccagggggac agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa
     4801 gggaatactg cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa
     4861 gcccgagaat atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa
     4921 gaacagtagg gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat
     4981 ccttaaggaa cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta
     5041 cctgcagaac ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga
     5101 ctacgacgtg gatcatatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa
     5161 agtgttgaca agatccgata aaaatagagg gaagagtgat aacgtcccct cagaagaagt
     5221 tgtcaagaaa atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg
     5281 gaagttcgat aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagccgg
     5341 cttcatcaaa aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct
     5401 cgattcacgc atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt
     5461 tattactctg aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt
     5521 gagagagatc aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac
     5581 tgcacttatc aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt
     5641 gtacgatgtt aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa
     5701 gtacttcttt tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg
     5761 agagattcgg aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga
     5821 caagggtagg gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt
     5881 taaaaagacc gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa
     5941 cagcgacaag ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga
     6001 ttctcctaca gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa
     6061 aaaactcaaa agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga
     6121 aaaaaacccc atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat
     6181 cattaagctt cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc
     6241 tagtgcgggc gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt
     6301 cttgtatctg gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa
     6361 gcagctgttc gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga
     6421 attctccaaa agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa
     6481 taagcacagg gataagccca tcagggagca ggcagaaaac attatccact tgtttactct
     6541 gaccaacttg ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg
     6601 gtacacctct acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct
     6661 ctatgaaaca agaatcgacc tctctcagct cggtggagac agcagggctg accccaagaa
     6721 gaagaggaag gtgtgatctc ttctcgagtc atgtaattag ttatgtcacg cttacattca
     6781 cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     6841 gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt
     6901 tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
     6961 gagaaggttt tgggacgctc gaaggcttta atttgcggcc ggtacccaat tcgccctata
     7021 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     7081 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     7141 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     7201 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     7261 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     7321 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     7381 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     7441 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     7501 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     7561 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     7621 cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg
     7681 catctgtgcg gtatttcaca ccgcatagat cggcaagtgc acaaacaata cttaaataaa
     7741 tactactcag taataaccta tttcttagca tttttgacga aatttgctat tttgttagag
     7801 tcttttacac catttgtctc cacacctccg cttacatcaa caccaataac gccatttaat
     7861 ctaagcgcat caccaacatt ttctggcgtc agtccaccag ctaacataaa atgtaagctt
     7921 tcggggctct cttgccttcc aacccagtca gaaatcgagt tccaatccaa aagttcacct
     7981 gtcccacctg cttctgaatc aaacaaggga ataaacgaat gaggtttctg tgaagctgca
     8041 ctgagtagta tgttgcagtc ttttggaaat acgagtcttt taataactgg caaaccgagg
     8101 aactcttggt attcttgcca cgactcatct ccatgcagtt ggacgatatc aatgccgtaa
     8161 tcattgacca gagccaaaac atcctcctta ggttgattac gaaacacgcc aaccaagtat
     8221 ttcggagtgc ctgaactatt tttatatgct tttacaagac ttgaaatttt ccttgcaata
     8281 accgggtcaa ttgttctctt tctattgggc acacatataa tacccagcaa gtcagcatcg
     8341 gaatctagag cacattctgc ggcctctgtg ctctgcaagc cgcaaacttt caccaatgga
     8401 ccagaactac ctgtgaaatt aataacagac atactccaag ctgcctttgt gtgcttaatc
     8461 acgtatactc acgtgctcaa tagtcaccaa tgccctccct cttggccctc tccttttctt
     8521 ttttcgaccg aattaattct taatcggcaa aaaaagaaaa gctccggatc aagattgtac
     8581 gtaaggtgac aagctatttt tcaataaaga atatcttcca ctactgccat ctggcgtcat
     8641 aactgcaaag tacacatata ttacgatgct gttctattaa atgcttccta tattatatat
     8701 atagtaatgt cgttgatcta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     8761 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     8821 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     8881 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     8941 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     9001 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     9061 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     9121 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     9181 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     9241 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     9301 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     9361 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     9421 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     9481 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     9541 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     9601 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     9661 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     9721 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     9781 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     9841 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     9901 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     9961 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
    10021 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
    10081 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
    10141 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
    10201 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
    10261 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
    10321 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
    10381 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
    10441 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
    10501 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
    10561 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
    10621 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
    10681 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
    10741 cctttttacg gttcctggcc ttttgctggc cttttgctcg ggggatctgc caattgaaca
    10801 taacatggta gttacatata ctagtaatat ggttcggcac acattaaaag tataaaaact
    10861 atctgaatta cgaattacat atattggtca taaaaatcaa tcaatcatcg tgtgttttat
    10921 atgtctctta tctaagtata agaatatcca tagttaatat tcacttacgc taccttttaa
    10981 cctgtaatca ttgtcaacag gatatgttaa cgacccacat tgataaacgc tagtatttct
    11041 ttttcctctt cttattggcc ggctgtctct atactcccct atagtctgtt tcttttcgtt
    11101 tcgattgtcc ctatcagtga tagagatggc gcacatggta cgctgtggtg ctcgcggctg
    11161 ggaacgaaac tctgggagct gcgattggca gtaaacacag atagatcaac agttttagag
    11221 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
    11281 tcggtgcttt tttctcgagc catatccaac ttccaattta atctttcttt tttaattttc
    11341 acttatttgc gatacagaaa gaaaaaagcg atagtaacta ttgaattttg tttggatttg
    11401 gttagattag atatggtttc tctttatatt tacatgctaa aaatgggcta caccagagat
    11461 acataattag atatatatac gccagtacac cttatcggcc caagccttgt cccaaggcag
    11521 cgttttgttc ttggaaacgc tgccctacac gttcgctatg cttcaagaac ttttctgagc
    11581 acttcatgat gcatgtttgt tccttattgg ttagctttga tgttgtgaag tcattgacac
    11641 agtctgtgaa acatctttct accagattag agtacaaacg catgaaatcc ttcatttgct
    11701 tttgttccac tactttttgg aactcttgtt gttctttcct gcaggtagtc gtgca
//
