LOCUS       sequence-330357-       11755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constituive spCas9 and tet-inducible CPR1 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_CPR1_1
FEATURES             Location/Qualifiers
     source          1..11755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             263..886
                     /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(336..355)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     1383..1400
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1517..1538
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1553..1558,1559..1576,1577..1583)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1591..1607
                     /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     1596..1618
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1615..1631
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1615..1631
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1650..1670
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1652..1670
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1688..2086
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     2090..2106
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2090..2106
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2118..6221
                     /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             2118..6221
                     /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             2118..6221
                     /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             6234..6254
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6270..6517
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6286..6304)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6535..6554)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6536..6554)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6564..6581)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6564..6580)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6573..6595)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6721..7176
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6808..6827)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7018..7039
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7279..7953)
                     /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(7954..8235)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(8256..8275)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8375..8397
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8435..8453)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8521..8625
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8626..8694,8695..9486)
                     /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(8844..8863)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9657..10245
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10146..10165
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    10629..10647
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        10733..10808
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      10821..10883
                     /label=RPR1 terminator
                     /note="transcription terminator for the S. cerevisiae RNase
                     P RNA gene"
ORIGIN
        1 tttcagctga cagcgagttt catgatcgtg atgaacaatg gtaacgagtt gtggctgttg
       61 ccagggaggg tggttctcaa cttttaatgt atggccaaat cgctacttgg gtttgttata
      121 taacaaagaa gaaataatga actgattctc ttcctccttc ttgtcctttc ttaattctgt
      181 tgtaattacc ttcctttgta attttttttg taattattct tcttaataat ccaaacaaac
      241 acacatatta caatatatta aaatgtctag attagataaa agtaaagtga ttaacagcgc
      301 attagagctg cttaatgagg tcggaatcga aggtttaaca acccgtaaac tcgcccagaa
      361 gctaggtgta gagcagccta cattgtattg gcatgtaaaa aataagcggg ctttgctcga
      421 cgccttagcc attgagatgt tagataggca ccatactcac ttttgccctt tagaagggga
      481 aagctggcaa gattttttac gtaataacgc taaaagtttt agatgtgctt tactaagtca
      541 tcgcgatgga gcaaaagtac atttaggtac acggcctaca gaaaaacagt atgaaactct
      601 cgaaaatcaa ttagcctttt tatgccaaca aggtttttca ctagagaatg cattatatgc
      661 actcagcgct gtggggcatt ttactttagg ttgcgtattg gaagatcaag agcatcaagt
      721 cgctaaagaa gaaagggaaa cacctactac tgatagtatg ccgccattat tacgacaagc
      781 tatcgaatta tttgatcacc aaggtgcaga gccagccttc ttattcggcc ttgaattgat
      841 catatgcgga ttagaaaaac aacttaaatg tgaaagtggg tcttaagtct gaagaatgaa
      901 tgatttgatg atttcttttt ccctccattt ttcttactga atatatcaat gatatagact
      961 tgtatagttt attatttcaa attaagtagc tatatatagt caagataacg tttgtttgac
     1021 acgattacat tattcgtcga catctttttt cagcctgtcg tggtagcaat ttgaggagta
     1081 ttattaattg aataggttca ttttgcgctc gcataaacag ttttcgtcag ggacagtatg
     1141 ttggaatgag tggtaattaa tggtgacatg acatgttata gcaataacct tgatgtttac
     1201 atcgtagttt aatgtacacc ccgcgaattc gttcaagtag gagtgcacca attgcaaagg
     1261 gaaaagctga atgggcagtt cgaaccgcgg tctttcctgc gttatcccct gattctgtgg
     1321 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     1381 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     1441 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     1501 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     1561 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     1621 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
     1681 tggagctcat agcttcaaaa tgtttctact ccttttttac tcttccagat tttctcggac
     1741 tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat ttcccctctt
     1801 tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa aaaagagacc
     1861 gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg tttctttttc
     1921 ttgaaaattt tttttttgat ttttttctct ttcgatgacc tcccattgat atttaagtta
     1981 ataaacggtc ttcaatttct caagtttcag tttcattttt cttgttctat tacaactttt
     2041 tttacttctt gctcattaga aagaaagcat agcaatctaa tctaagtttt ctagaactag
     2101 tggatccccc gggaaaaatg gacaagaagt actccattgg gctcgatatc ggcacaaaca
     2161 gcgtcggttg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc
     2221 tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc ctgttcgact
     2281 ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga tatacccgca
     2341 gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct aaggtggatg
     2401 actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa aagcacgagc
     2461 gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag tacccaacca
     2521 tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg cggttgatct
     2581 atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag ggggacctga
     2641 acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact tacaatcagc
     2701 ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc ctgagcgcta
     2761 ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga
     2821 acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac tttaaatcta
     2881 acttcgacct ggccgaagat gccaagcttc aactgagcaa agacacctac gatgatgatc
     2941 tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg gcggcaaaga
     3001 acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag atcaccaaag
     3061 ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac ttgactttgc
     3121 tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt
     3181 ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat
     3241 ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta aagcttaaca
     3301 gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc
     3361 acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag
     3421 ataacaggga aaagattgag aaaatcctca catttcggat accctactat gtaggccccc
     3481 tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag accatcactc
     3541 cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga
     3601 tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt
     3661 acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga
     3721 gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga
     3781 cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag attgaatgtt
     3841 tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc
     3901 acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca
     3961 ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac
     4021 gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc
     4081 gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga gacaagcaga
     4141 gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc
     4201 agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg
     4261 gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa
     4321 agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata
     4381 agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga
     4441 agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa
     4501 tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact
     4561 acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat cggctctccg
     4621 actacgacgt ggatcatatc gtgccccagt cttttctcaa agatgattct attgataata
     4681 aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag
     4741 ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac
     4801 ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg
     4861 gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc
     4921 tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag
     4981 ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag ttttataagg
     5041 tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca
     5101 ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga gactataaag
     5161 tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta
     5221 agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg
     5281 gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg
     5341 acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag gtgaacatcg
     5401 ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga
     5461 acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg
     5521 attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta
     5581 aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga tcaagcttcg
     5641 aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa aaagacctca
     5701 tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg
     5761 ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt
     5821 tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga
     5881 agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg
     5941 aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca
     6001 ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac ttgtttactc
     6061 tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaagc
     6121 ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc
     6181 tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct gaccccaaga
     6241 agaagaggaa ggtgtgatct cttctcgagt catgtaatta gttatgtcac gcttacattc
     6301 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     6361 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat
     6421 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct
     6481 tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc cggtacccaa ttcgccctat
     6541 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     6601 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     6661 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     6721 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     6781 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     6841 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     6901 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     6961 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     7021 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     7081 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     7141 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     7201 gcatctgtgc ggtatttcac accgcataga tcggcaagtg cacaaacaat acttaaataa
     7261 atactactca gtaataacct atttcttagc atttttgacg aaatttgcta ttttgttaga
     7321 gtcttttaca ccatttgtct ccacacctcc gcttacatca acaccaataa cgccatttaa
     7381 tctaagcgca tcaccaacat tttctggcgt cagtccacca gctaacataa aatgtaagct
     7441 ttcggggctc tcttgccttc caacccagtc agaaatcgag ttccaatcca aaagttcacc
     7501 tgtcccacct gcttctgaat caaacaaggg aataaacgaa tgaggtttct gtgaagctgc
     7561 actgagtagt atgttgcagt cttttggaaa tacgagtctt ttaataactg gcaaaccgag
     7621 gaactcttgg tattcttgcc acgactcatc tccatgcagt tggacgatat caatgccgta
     7681 atcattgacc agagccaaaa catcctcctt aggttgatta cgaaacacgc caaccaagta
     7741 tttcggagtg cctgaactat ttttatatgc ttttacaaga cttgaaattt tccttgcaat
     7801 aaccgggtca attgttctct ttctattggg cacacatata atacccagca agtcagcatc
     7861 ggaatctaga gcacattctg cggcctctgt gctctgcaag ccgcaaactt tcaccaatgg
     7921 accagaacta cctgtgaaat taataacaga catactccaa gctgcctttg tgtgcttaat
     7981 cacgtatact cacgtgctca atagtcacca atgccctccc tcttggccct ctccttttct
     8041 tttttcgacc gaattaattc ttaatcggca aaaaaagaaa agctccggat caagattgta
     8101 cgtaaggtga caagctattt ttcaataaag aatatcttcc actactgcca tctggcgtca
     8161 taactgcaaa gtacacatat attacgatgc tgttctatta aatgcttcct atattatata
     8221 tatagtaatg tcgttgatct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     8281 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     8341 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     8401 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     8461 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     8521 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     8581 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     8641 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     8701 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     8761 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     8821 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     8881 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     8941 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     9001 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     9061 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     9121 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     9181 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     9241 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     9301 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     9361 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     9421 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     9481 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     9541 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     9601 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     9661 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     9721 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     9781 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     9841 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     9901 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     9961 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
    10021 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
    10081 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
    10141 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
    10201 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
    10261 gcctttttac ggttcctggc cttttgctgg ccttttgctc gggggatctg ccaattgaac
    10321 ataacatggt agttacatat actagtaata tggttcggca cacattaaaa gtataaaaac
    10381 tatctgaatt acgaattaca tatattggtc ataaaaatca atcaatcatc gtgtgtttta
    10441 tatgtctctt atctaagtat aagaatatcc atagttaata ttcacttacg ctacctttta
    10501 acctgtaatc attgtcaaca ggatatgtta acgacccaca ttgataaacg ctagtatttc
    10561 tttttcctct tcttattggc cggctgtctc tatactcccc tatagtctgt ttcttttcgt
    10621 ttcgattgtc cctatcagtg atagagatgg cgcacatggt acgctgtggt gctcgcggct
    10681 gggaacgaaa ctctgggagc tgcgattggc agggcggtaa gtctatctac gggttttaga
    10741 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
    10801 gtcggtgctt ttttctcgag ccatatccaa cttccaattt aatctttctt ttttaatttt
    10861 cacttatttg cgatacagaa agaaaaaagc gatagtaact attgaatttt gtttggattt
    10921 ggttagatta gatatggttt ctctttatat ttacatgcta aaaatgggct acaccagaga
    10981 tacataatta gatatatata cgccagtaca ccttatcggc ccaagccttg tcccaaggca
    11041 gcgttttgtt cttggaaacg ctgccctaca cgttcgctat gcttcaagaa cttttctgag
    11101 cacttcatga tgcatgtttg ttccttattg gttagctttg atgttgtgaa gtcattgaca
    11161 cagtctgtga aacatctttc taccagatta gagtacaaac gcatgaaatc cttcatttgc
    11221 ttttgttcca ctactttttg gaactcttgt tgttctttcc tgcaggtagt cgtgcaatgt
    11281 atgactttaa gatttgtgag caggaagaaa agggagaatc ttctaacgat aaacccttga
    11341 aaaactgggt agactacgct atgttgagtt gctacgcagg ctgcacaatt acacgagaat
    11401 gctcccgcct aggatttaag gctaagggac gtgcaatgca gacgacagat ctaaatgacc
    11461 gtgtcggtga agtgttcgcc aaacttttcg gttaacacat gcagtgatgc acgcgcgatg
    11521 gtgctaagtt acatatatat atatatgtat agccatagtg atgtctaagt aacctttatg
    11581 gtatatttct taatgtggaa agatactagc gcgcgcaccc acacacaagc ttcgtctttt
    11641 cttgaagaaa agaggaagct cgctaaatgg gattccactt tccgttccct gccagctgat
    11701 ggaaaaaggt tagtggaacg atgaagaata aaaagagaga tccactgagg tgaaa
//
