LOCUS       sequence-212701-       14966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Solo vector pV1382 + sgCgADE2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14966)
  AUTHORS   Vyas VK, Bushkin GG, Bernstein DA, Getz MA, Sewastianik M, Barrasa 
            MI, Bartel DP, Fink GR
  TITLE     New CRISPR Mutagenesis Strategies Reveal Variation in Repair 
            Mechanisms among Fungi.
  JOURNAL   mSphere. 2018 Apr 25;3(2). pii: 3/2/e00154-18. doi: 
            10.1128/mSphere.00154-18. Print 2018 Apr 25.
  PUBMED    29695624
REFERENCE   2  (bases 1 to 14966)
  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"; journalName: "mSphere."; 
            date: "2018-04-25"; volume: "3(2). pii: 3/2/e00154-18. doi"; pages: 
            " 10.1128/mSphere.00154-18. Print 2018 Apr 25"
COMMENT     Sequence Label: pV1435
FEATURES             Location/Qualifiers
     source          1..14966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(33..55)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     158..177
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        201..416
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             417..1220
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1351..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1488..1509)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1700..1719
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1932..1954
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1946..1963
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1947..1963
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1973..1992
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1973..1991
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3579..3992
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             4012..8115
                     /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             4012..8115
                     /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             8128..8148
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8152..8172
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8176..8196
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8197..8262
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(8343..8361)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      complement(8519..8716)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(9317..9660)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        9708..9976
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        9998..10073
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(12193..12213)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12193..12211)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12232..12248)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12232..12248)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12245..12267)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12256..12272
                     /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)."
     promoter        complement(join(12280..12286,12287..12304,12305..12310))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12325..12346
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12463..12480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12634..13222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12714..12733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13393..14184,14185..14253))
                     /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     14016..14035
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14254..14358)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    14395..14898
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     14939..14957
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gaattcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagatc
       61 tcggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      121 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtaccg
      181 agagtgcacc ataccacctt ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggtgatg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     1861 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     1921 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
     1981 ctcactatag ggcgaattgg gtacctgcca ctactaccac tgggagtttc gttcttctcg
     2041 atactattag ctttacttcc tgcactagca gtggttggat caacagaatc ttcataatca
     2101 tcaaaatcgt cttttgaaga ccccccgttt gatgtatggc cctgtctttt catcaaactt
     2161 tttatatagt tgactgaact gaggctaaat atgtgatcat cttcactata gacaatcttt
     2221 ctcttatttg caccaccgcc accaccagtc tttgagaaat tctcaaaacc ttttacgata
     2281 ttaccaagcg ggctctcttc gaaataatct atctcttttt gatatatcga atcctctagc
     2341 gtggttagct ttctagttag ttcttgcttc ttaagaattt gctggattag tttatttttc
     2401 aattcaacgt atttctcaga gtcatcttta gattttgatg aagatgtgcg ttcattcgct
     2461 atatccttct tggtcgtgtc ttttcgatcc tccttggctg gcactgaact cgtctttttt
     2521 ggcgttgctg ttccagacag acttatctca ttagatttgg aacttgtggg tttaacatca
     2581 tttgcatctt tagtagacat gattgtgtaa taccgtgatt gtttgttttg aaaggtctgt
     2641 catatttcta tcaatttcaa aacaaaatgt tcatcagaaa aaagccaaaa atgtctcttc
     2701 tagcttctta gtggtgtcgc aaaatacaca atgtcgctca acaatccaca ttcccggcgc
     2761 atagctcaaa tcacatgact acagctaaca attacacaaa aaaattctct tttcgatgta
     2821 gcaactatct tcaactaaaa cattttctcc ttcggcccat gattgtcctc cgggtcgaca
     2881 gcaagccgtt acaattgaga tggaaagcga cctaccttca ctcgataagg tgcttaattg
     2941 tacttcatat aaatctggcc cggatctaaa caaatgagtt ccattaagcc gtgggttctc
     3001 aattagggtt tttgtttttg atttagaaaa aagagatcaa gatttgttta caggtgatgc
     3061 ctttttttag aacttatgcg ttgcaaaagt tgactaacga tttctataag gtgatccaca
     3121 ctaattatac aaacgtacaa acagacatac ttttcctgcg ttcacctgat gttggccaga
     3181 tttctctctt cattgcatag aacataacca cactagggca acagaaaaaa aaaaaaaaag
     3241 tgcatcggga agttgtgttc cattcattat atgtctacta ctgcatatga gtagcccacc
     3301 caccaccacc atagtaagtt tttgtgtatg cgcgccgtca ggttatttca tttctgaatt
     3361 tttcaaccac cttactccct ttattgttga ttgacaattt tgctcacagt aagatctttt
     3421 agactccaat taatataaaa taagtctgat tttccaattc ctgttttttc tttttttttc
     3481 tgtttctatt tctttccttt tctccctttt ttttaattct tcattcaatc atcaattgat
     3541 aattcaggaa tattacaacc accatggccc ccacaagtga tcccccacac accatagctt
     3601 caaaatgttt ctactccttt tttactcttc cagattttct cggactccgc gcatcgccgt
     3661 accacttcaa aacacccaag cacagcatac taaatttccc ctctttcttc ctctagggtg
     3721 tcgttaatta cccgtactaa aggtttggaa aagaaaaaag agaccgcctc gtttcttttt
     3781 cttcgtcgaa aaaggcaata aaaattttta tcacgtttct ttttcttgaa aatttttttt
     3841 tttgattttt ttctctttcg atgacctccc attgatattt aagttaataa acggtcttca
     3901 atttctcaag tttcagtttc atttttcttg ttctattaca acttttttta cttcttgctc
     3961 attagaaaga aagcatagca atctaatcta agttttaatt acaaacctgg gatggataaa
     4021 aagtatagta ttggtttaga tattggtact aactctgtgg gttgggcagt tatcactgac
     4081 gaatataaag ttccatcaaa gaaatttaag gtgttaggta acactgacag acactcaata
     4141 aaaaagaatc ttatcggtgc tcttttgttc gactccggtg aaactgccga ggctacacgt
     4201 ttaaaaagaa cagcaagaag aagatatacc cgtagaaaaa atagaatatg ttatttacaa
     4261 gaaatctttt ctaatgaaat ggctaaagtt gatgattcct ttttccatag attggaagag
     4321 tcatttttgg ttgaagaaga caaaaagcat gagagacatc caatctttgg gaatatagtt
     4381 gatgaagtgg cttaccatga aaaatatcct accatttatc atttaagaaa gaaattggta
     4441 gattcaactg ataaagctga ccttagatta atctatttag cacttgccca tatgattaaa
     4501 tttagaggtc attttttgat tgaaggtgat ttgaacccag ataattctga cgtggataaa
     4561 ttatttattc aattagtcca aacctacaac caattatttg aggaaaatcc aattaatgct
     4621 agtggtgtcg atgccaaagc tatattatca gccagattat caaaatctag acgtttggaa
     4681 aatttgattg cccaattgcc aggagaaaaa aagaatggat tatttggaaa cttgatcgca
     4741 ttatcattgg gtttgacacc aaattttaaa tctaattttg atttagctga agatgctaaa
     4801 ttacaattat caaaagacac ctatgacgac gatttggaca atttacttgc tcaaattggt
     4861 gatcaatatg cagatttgtt cttagctgct aaaaacttat ctgatgctat tttgttgtct
     4921 gatattttga gagtgaacac agaaataacc aaagctccat tatcagcatc tatgatcaaa
     4981 cgttatgatg aacaccatca ggatttgact ttattgaaag ctttggtgag acaacaattg
     5041 ccagagaagt ataaagaaat ctttttcgat caatctaaaa acgggtatgc aggttatatt
     5101 gatgggggtg cctcccaaga ggaattttac aaatttataa aacctatttt agaaaagatg
     5161 gatgggactg aggaactttt ggtcaaattg aacagagaag atttgttacg taaacagaga
     5221 acttttgata atggtagtat acctcaccaa attcatttgg gtgagttgca tgcaatttta
     5281 agaagacaag aagattttta tccattttta aaagataata gagaaaaaat cgagaaaatt
     5341 ttaaccttta gaattccata ctatgttggg cctttggcta gaggtaattc aagatttgcc
     5401 tggatgacac gtaaatcaga agaaactatt accccttgga attttgaaga ggttgttgat
     5461 aaaggagcat cagcacagag ttttattgaa agaatgacca atttcgataa aaacttacca
     5521 aatgaaaaag ttttaccaaa acattccttg ttatacgaat attttactgt ttacaatgaa
     5581 cttacaaagg ttaaatatgt tactgaaggt atgcgtaagc cagccttttt atctggagaa
     5641 cagaaaaagg caatagttga tttattgttt aaaacaaata gaaaagttac tgttaaacaa
     5701 ttaaaagaag attactttaa gaaaattgaa tgttttgatt cagttgaaat cagtggtgtt
     5761 gaagacagat ttaatgctag tttaggaact taccatgatt tacttaaaat tatcaaagat
     5821 aaagatttct tggataacga agaaaatgaa gacattttag aagacattgt tttaacctta
     5881 actttattcg aagatagaga gatgattgaa gaacgtttga agacttatgc acatttgttt
     5941 gacgataaag tgatgaaaca gttgaaaaga agacgttata ctggatgggg tagattgtct
     6001 cgtaaattga tcaatggaat tagagataaa caaagtggta aaactatctt ggactttttg
     6061 aaatctgacg gatttgctaa tagaaatttc atgcaattga tccacgacga tagtttgaca
     6121 tttaaagaag acatccaaaa ggcccaagtg agtgggcaag gtgattcatt acatgaacat
     6181 attgcaaatt tagccggatc tcctgctatt aagaaaggga tattacaaac tgttaaagtt
     6241 gtggatgaat tagtgaaagt aatgggaaga cataaacctg aaaacattgt cattgagatg
     6301 gcaagagaaa atcaaactac acaaaaagga cagaaaaata gtagagaacg tatgaaaaga
     6361 atagaagagg gtattaaaga attgggtagt caaatattga aagaacaccc agtggaaaat
     6421 acccagttgc aaaatgaaaa attatatctt tactaccttc aaaatggacg tgatatgtat
     6481 gttgatcagg aattagatat aaatagactt tcagattatg atgtagatca tatagttcca
     6541 caatctttct tgaaagatga ttccatagac aataaagtat taactagaag tgataaaaat
     6601 agaggtaaaa gtgataatgt cccaagtgag gaagtcgtca aaaagatgaa aaattactgg
     6661 cgtcaacttt tgaatgctaa attaattact caaagaaaat ttgataattt gactaaagca
     6721 gaaagaggtg ggctttctga attagataaa gccgggttca ttaaaagaca attggtcgaa
     6781 actagacaaa ttactaaaca tgttgcccaa attttagatt cccgtatgaa cactaagtat
     6841 gacgaaaatg ataagttaat acgtgaggtt aaagtcatta ctttaaaatc aaaacttgtc
     6901 tctgatttca gaaaggattt ccaattctat aaagttagag aaattaataa ttatcatcat
     6961 gctcatgatg catatttgaa tgctgtagtt ggaactgctt taatcaagaa ataccctaaa
     7021 ttagaatctg aatttgtata tggtgattac aaagtctatg atgttagaaa gatgattgct
     7081 aaatcagaac aagaaattgg taaagctaca gctaaatact tcttttactc taacattatg
     7141 aatttcttta aaacagaaat tactttggca aacggtgaaa ttagaaaaag acctcttatt
     7201 gaaacaaatg gtgagactgg agagatagtt tgggacaaag ggcgtgattt cgctactgtt
     7261 agaaaagttt tatcaatgcc acaagttaac attgtaaaga aaacagaggt tcaaactggt
     7321 ggtttctcaa aagaaagtat tttgcctaaa agaaatagtg ataaattgat tgccagaaaa
     7381 aaggattggg atccaaagaa atatggtggt ttcgactcac caaccgtagc ctattctgtt
     7441 ttggttgtgg caaaggttga aaagggtaaa agtaaaaagc ttaaatcagt aaaagaactt
     7501 ttgggtatta caataatgga aagaagttcc tttgaaaaga accctattga ttttttggaa
     7561 gctaaaggtt ataaggaagt aaagaaggac ttaataatca aattgcctaa atattcttta
     7621 tttgaattag aaaatgggag aaaaagaatg ttggcttctg ctggagaatt gcaaaagggt
     7681 aatgaattag cattgccttc caaatatgtt aacttcttgt atttagcttc acactatgaa
     7741 aagttgaaag ggtcaccaga agataacgag caaaaacaat tatttgttga acaacacaaa
     7801 cactacttag atgagattat agaacaaatt agtgaattca gtaaaagagt gatattagct
     7861 gatgcaaatt tagataaagt tttgtcagcc tataacaaac atagagataa gccaattaga
     7921 gaacaagcag aaaacattat tcacttattt acccttacca atttaggagc acctgctgct
     7981 ttcaagtatt ttgatacaac aattgatcgt aaaagatata cctcaacaaa agaagtctta
     8041 gacgccacct taattcatca atcaatcact ggattgtatg agacaagaat tgatttgtct
     8101 caattgggtg gtgatgaagg ggctgatcct aagaagaaaa gaaaagttga tccaaagaaa
     8161 aagcgtaagg tggatcctaa gaaaaagaga aaggttgact acaaagacca tgacggtgat
     8221 tataaagatc atgacatcga ctacaaggat gacgatgaca agtgataatg actgcagaga
     8281 tccatcgacc tgccgccaag ctaattccgg gcgaatttct gtcgagtcat gtaattagtt
     8341 atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt
     8401 agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg
     8461 ttatttatat ttcaaatttt tctttttttt cttggcggcg ttagtatcaa atcgacagca
     8521 gtatagcgac cagcattcac atacgattga cgcatgatat tactttctgc gcacttaact
     8581 tcgcatctgg gcagatgatg tcgaggcgaa aaaaaatata aatcacgcta acatttgatt
     8641 aaaatagaac aactacaata taaaaaaact atacaaatga caagttcttg aaaacaagaa
     8701 tctttttatt gtcagtactg actcgagtta ttatggacat ggcatagaca tatacaaagc
     8761 ttgttcacca tcggaagcag taccatcgta taaagcagta tccaaaccac acaaagtgaa
     8821 acccattctt ctataagcat gaatagctgg agcattaaca ttggtaactt ccaaccacaa
     8881 atgaccagca cctctttctc tggcgaattc agtagccaaa cccatcaaag ctctaccaac
     8941 accatgacct ctatgttctg gagcaacttc aatatcttca acagtcaatc ttctgttcca
     9001 accagaataa gaaacaacaa cgaaaccagc caaatcacca tcatcaccat aagcaacgaa
     9061 agttctagaa tctggatcac catcttcacc agcatcggat tcatcatcgg attcatcatc
     9121 tgggaaaacc ttagtcaatg gtggatcaac tggaacttct ctcaaagtga aaccatcacc
     9181 agtagcagta actctaaaaa cagtatcggt agtgaaagaa ccatccaaag cttcaatagc
     9241 ttcagcatca cctggaacag aagttctgta tctataagca gtatcatcca aagtagtaga
     9301 cataattgta ggatccggtt gtttatgttc ggatgtgatg tgagaactgt atcctagcaa
     9361 gattttaaaa ggaagtatat gaaagaagaa cctcagtggc aaatcctaac cttttatatt
     9421 tctctacagg ggcgcggcgt ggggacaatt caacgcgtct gtgaggggag cgtttccctg
     9481 ctcgcaggtc tgcagcgagg agccgtaatt tttgcttcgc gccgtgcggc catcaaaatg
     9541 tatggatgca aatgattata catggggatg tatgggctaa atgtacgggc gacagtcaca
     9601 tcatgcccct gagctgcgca cgtcaagact gtcaaggagg gtattctggg cctccatgtc
     9661 gctggccggg tgacccggcg gggacgaggc aagcttgatg cggccgctct ttgaaaagat
     9721 aatgtatgat tttgctttca ctcatattta tacagaaact tgatgttttc tttcgagtat
     9781 atacaaggtg attacatgta cgtttgaagt acaactctag attttgtagt gccctcttgg
     9841 gctagcggta aaggtgcgca ttttttcaca ccctacaatg ttctgttcaa aagattttgg
     9901 tcaaacgctg tagaagtgaa agttggtgcg catgtttcgg cgttcgaaac ttctccgcag
     9961 tgaaagataa atgatcggtt gggccgtatg attgtcggtt ttagagctag aaatagcaag
    10021 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt
    10081 ctcgagtttt tttatcgagt gtttaaggat aatgataact gaagagaaga attagttttg
    10141 ccgccaccgc gggtttgcct ctgattaaat aaaaaaaagc tggtgctttt ttttctttta
    10201 taggaacatc ttgaatatat gaactaatta aatgataatt ttttacccat ctttactctt
    10261 aatcactgag ctgcagtcaa agaaaaaggg atacagcacc tggtgaagag atgaacggag
    10321 actaacttag acgcgttgat tcttttcaat tgcacatttt attaatcgat gctaacgtct
    10381 atttacatat attctttaga gatattatct agggcttcaa ataatctctg gacagcaata
    10441 aaagtctctt caaaagtatt gtataacggc aatggggcta atctgattac atctggtctt
    10501 ctttcgtcac agattatagc atgatcatgc aagtacgcat taactcgttc catgacgttc
    10561 ttgccctttt catcgaaatg cggttgaaac ataatggaca attgacatcc tctttcagct
    10621 ggattcaaag gagttaaaat tttaaaccca aatttggagt ttgatgtact ggattgtggt
    10681 atgtaatact tggaattcgt caatagatcc tgtaaaaatt gagtcaaagc aacacttttt
    10741 tcacgaagtt tagatactcc acccacttta gcatacactt ccaatgacga cttcacagca
    10801 acaacatcaa gaacagaagg atttgactgt ctgtaagaaa gagccgagtt tattggatca
    10861 aactcttcta acattttgaa tcgttcttgg gagttattgc cccaccaacc agctagtcta
    10921 ggaacgaaac tgctcttctt gttctctata gtgtattttt catgcacaaa aattccacct
    10981 atggctccag gtcccgagtt taaatatttg taggaacacc aagcagcaaa atctactccc
    11041 caatcatgta aatttaatgg aacattccca actgcatggg caagatccca cccaacttta
    11101 atttgttggc tcttttcctt agcgtattta gttatttcct ctatcttgaa aaattgacca
    11161 gtgtagtatt ggataccagg aaaacacact agagccaatt catccaggtt ctcatctata
    11221 gccttgatta ttctttctgt tttaatataa gtttcaccag gttgaacttc caattgaatc
    11281 aaatgtttct cgtcgtatcc gaacaattta acaatgttca aaaatgcata gtagtcagaa
    11341 ggaaatgctt gtttttcaaa taaaattttg gttcttttcc cctcaggttt gtaaaaatgg
    11401 atcaacaatg cattcaagtt tgctgttaaa gaacccataa ctgcaacttc gttttccttt
    11461 gcaccaacaa tgggggctat taatggtaat aagggtaaat cgatgtctac ccacggtgtt
    11521 aacagtttgt caggatgatt gaaatgagac tcaacccctc gttcaaccca tgcattcaat
    11581 tcatcattga tagctttctt tgtattctta ggcatcaacc caagagagtt tccacataaa
    11641 taaatagact cagttgatga ctcatattta ttatttttga tacctaatga tccaaaagtt
    11701 ggtatggcaa actcattttt aaaagttggg aactttttgt ccaatttctt tgcctcggct
    11761 aatgacatct gataataaaa tggggttgga gtagttggtg gtataaccgg agagatagaa
    11821 ttgaagaaaa aaatcggaaa caacaaaaaa agttgatacc cagtattatg tgggagataa
    11881 ttgcgaatgg tggaaaaaaa aaagacgcct ttgggcctca acaacaattc taacagcttg
    11941 agagctttac aatcgagaaa ctatgattca ttccgtttta atatgtatgt acttagtaaa
    12001 ctcgtgaatt ttatttgtgg tctactttag tactaacata atcattggat agtcaataat
    12061 gatggtcttc cgagactaat gaaattctat accaaagtcg atattccaac acagaaattg
    12121 ctcttgcaac aagtgcacct gttgatatct actagatcag cacaagagtg ttcggagctc
    12181 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
    12241 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
    12301 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
    12361 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
    12421 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
    12481 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
    12541 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
    12601 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    12661 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    12721 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    12781 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    12841 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    12901 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    12961 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    13021 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
    13081 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    13141 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    13201 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
    13261 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
    13321 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
    13381 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
    13441 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    13501 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
    13561 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
    13621 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    13681 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
    13741 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
    13801 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
    13861 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
    13921 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
    13981 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
    14041 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
    14101 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
    14161 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
    14221 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
    14281 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
    14341 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcct tttcatcacg
    14401 tgctataaaa ataattataa tttaaatttt ttaatataaa tatataaatt aaaaatagaa
    14461 agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc cgaagatgta aaagactcta
    14521 gggggatcgc caacaaatac taccttttat cttgctcttc ctgctctcag gtattaatgc
    14581 cgaattgttt catcttgtct gtgtagaaga ccacacacga aaatcctgtg attttacatt
    14641 ttacttatcg ttaatcgaat gtatatctat ttaatctgct tttcttgtct aataaatata
    14701 tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt gtttattttt ttttcttcat
    14761 tccgtaactc ttctaccttc tttatttact ttctaaaatc caaatacaaa acataaaaat
    14821 aaataaacac agagtaaatt cccaaattat tccatcatta aaagatacga ggcgcgtgta
    14881 agttacaggc aagcgatccg tctaagaaac cattattatc atgacattaa cctataaaaa
    14941 taggcgtatc acgaggccct ttcgtc
//
