LOCUS       sequence-212593-       14965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  S. cerevisiae and C. glabrata Solo CRISPR vector, marked with URA3 
            and NatR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14965)
  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 14965)
  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: pV1382
FEATURES             Location/Qualifiers
     source          1..14965
                     /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..3993
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             4013..8116
                     /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             4013..8116
                     /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             8129..8149
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8153..8173
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8177..8197
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8198..8263
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(8344..8362)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      complement(8520..8717)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(9318..9661)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        9709..9977
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        9997..10072
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(12192..12212)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12192..12210)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12231..12247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12231..12247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12244..12266)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12255..12271
                     /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(12279..12285,12286..12303,12304..12309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12324..12345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12462..12479)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12633..13221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12713..12732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13392..14183,14184..14252))
                     /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     14015..14034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14253..14357)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    14394..14897
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     14938..14956
                     /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 aacaattgat
     3541 aattcaggaa tattacaaca accatggccc ccacaagtga tcccccacac acaccatagc
     3601 ttcaaaatgt ttctactcct tttttactct tccagatttt ctcggactcc gcgcatcgcc
     3661 gtaccacttc aaaacaccca agcacagcat actaaatttc ccctctttct tcctctaggg
     3721 tgtcgttaat tacccgtact aaaggtttgg aaaagaaaaa agagaccgcc tcgtttcttt
     3781 ttcttcgtcg aaaaaggcaa taaaaatttt tatcacgttt ctttttcttg aaaatttttt
     3841 ttttgatttt tttctctttc gatgacctcc cattgatatt taagttaata aacggtcttc
     3901 aatttctcaa gtttcagttt catttttctt gttctattac aacttttttt acttcttgct
     3961 cattagaaag aaagcatagc aatctaatct aagttttaat tacaaacctg ggatggataa
     4021 aaagtatagt attggtttag atattggtac taactctgtg ggttgggcag ttatcactga
     4081 cgaatataaa gttccatcaa agaaatttaa ggtgttaggt aacactgaca gacactcaat
     4141 aaaaaagaat cttatcggtg ctcttttgtt cgactccggt gaaactgccg aggctacacg
     4201 tttaaaaaga acagcaagaa gaagatatac ccgtagaaaa aatagaatat gttatttaca
     4261 agaaatcttt tctaatgaaa tggctaaagt tgatgattcc tttttccata gattggaaga
     4321 gtcatttttg gttgaagaag acaaaaagca tgagagacat ccaatctttg ggaatatagt
     4381 tgatgaagtg gcttaccatg aaaaatatcc taccatttat catttaagaa agaaattggt
     4441 agattcaact gataaagctg accttagatt aatctattta gcacttgccc atatgattaa
     4501 atttagaggt cattttttga ttgaaggtga tttgaaccca gataattctg acgtggataa
     4561 attatttatt caattagtcc aaacctacaa ccaattattt gaggaaaatc caattaatgc
     4621 tagtggtgtc gatgccaaag ctatattatc agccagatta tcaaaatcta gacgtttgga
     4681 aaatttgatt gcccaattgc caggagaaaa aaagaatgga ttatttggaa acttgatcgc
     4741 attatcattg ggtttgacac caaattttaa atctaatttt gatttagctg aagatgctaa
     4801 attacaatta tcaaaagaca cctatgacga cgatttggac aatttacttg ctcaaattgg
     4861 tgatcaatat gcagatttgt tcttagctgc taaaaactta tctgatgcta ttttgttgtc
     4921 tgatattttg agagtgaaca cagaaataac caaagctcca ttatcagcat ctatgatcaa
     4981 acgttatgat gaacaccatc aggatttgac tttattgaaa gctttggtga gacaacaatt
     5041 gccagagaag tataaagaaa tctttttcga tcaatctaaa aacgggtatg caggttatat
     5101 tgatgggggt gcctcccaag aggaatttta caaatttata aaacctattt tagaaaagat
     5161 ggatgggact gaggaacttt tggtcaaatt gaacagagaa gatttgttac gtaaacagag
     5221 aacttttgat aatggtagta tacctcacca aattcatttg ggtgagttgc atgcaatttt
     5281 aagaagacaa gaagattttt atccattttt aaaagataat agagaaaaaa tcgagaaaat
     5341 tttaaccttt agaattccat actatgttgg gcctttggct agaggtaatt caagatttgc
     5401 ctggatgaca cgtaaatcag aagaaactat taccccttgg aattttgaag aggttgttga
     5461 taaaggagca tcagcacaga gttttattga aagaatgacc aatttcgata aaaacttacc
     5521 aaatgaaaaa gttttaccaa aacattcctt gttatacgaa tattttactg tttacaatga
     5581 acttacaaag gttaaatatg ttactgaagg tatgcgtaag ccagcctttt tatctggaga
     5641 acagaaaaag gcaatagttg atttattgtt taaaacaaat agaaaagtta ctgttaaaca
     5701 attaaaagaa gattacttta agaaaattga atgttttgat tcagttgaaa tcagtggtgt
     5761 tgaagacaga tttaatgcta gtttaggaac ttaccatgat ttacttaaaa ttatcaaaga
     5821 taaagatttc ttggataacg aagaaaatga agacatttta gaagacattg ttttaacctt
     5881 aactttattc gaagatagag agatgattga agaacgtttg aagacttatg cacatttgtt
     5941 tgacgataaa gtgatgaaac agttgaaaag aagacgttat actggatggg gtagattgtc
     6001 tcgtaaattg atcaatggaa ttagagataa acaaagtggt aaaactatct tggacttttt
     6061 gaaatctgac ggatttgcta atagaaattt catgcaattg atccacgacg atagtttgac
     6121 atttaaagaa gacatccaaa aggcccaagt gagtgggcaa ggtgattcat tacatgaaca
     6181 tattgcaaat ttagccggat ctcctgctat taagaaaggg atattacaaa ctgttaaagt
     6241 tgtggatgaa ttagtgaaag taatgggaag acataaacct gaaaacattg tcattgagat
     6301 ggcaagagaa aatcaaacta cacaaaaagg acagaaaaat agtagagaac gtatgaaaag
     6361 aatagaagag ggtattaaag aattgggtag tcaaatattg aaagaacacc cagtggaaaa
     6421 tacccagttg caaaatgaaa aattatatct ttactacctt caaaatggac gtgatatgta
     6481 tgttgatcag gaattagata taaatagact ttcagattat gatgtagatc atatagttcc
     6541 acaatctttc ttgaaagatg attccataga caataaagta ttaactagaa gtgataaaaa
     6601 tagaggtaaa agtgataatg tcccaagtga ggaagtcgtc aaaaagatga aaaattactg
     6661 gcgtcaactt ttgaatgcta aattaattac tcaaagaaaa tttgataatt tgactaaagc
     6721 agaaagaggt gggctttctg aattagataa agccgggttc attaaaagac aattggtcga
     6781 aactagacaa attactaaac atgttgccca aattttagat tcccgtatga acactaagta
     6841 tgacgaaaat gataagttaa tacgtgaggt taaagtcatt actttaaaat caaaacttgt
     6901 ctctgatttc agaaaggatt tccaattcta taaagttaga gaaattaata attatcatca
     6961 tgctcatgat gcatatttga atgctgtagt tggaactgct ttaatcaaga aataccctaa
     7021 attagaatct gaatttgtat atggtgatta caaagtctat gatgttagaa agatgattgc
     7081 taaatcagaa caagaaattg gtaaagctac agctaaatac ttcttttact ctaacattat
     7141 gaatttcttt aaaacagaaa ttactttggc aaacggtgaa attagaaaaa gacctcttat
     7201 tgaaacaaat ggtgagactg gagagatagt ttgggacaaa gggcgtgatt tcgctactgt
     7261 tagaaaagtt ttatcaatgc cacaagttaa cattgtaaag aaaacagagg ttcaaactgg
     7321 tggtttctca aaagaaagta ttttgcctaa aagaaatagt gataaattga ttgccagaaa
     7381 aaaggattgg gatccaaaga aatatggtgg tttcgactca ccaaccgtag cctattctgt
     7441 tttggttgtg gcaaaggttg aaaagggtaa aagtaaaaag cttaaatcag taaaagaact
     7501 tttgggtatt acaataatgg aaagaagttc ctttgaaaag aaccctattg attttttgga
     7561 agctaaaggt tataaggaag taaagaagga cttaataatc aaattgccta aatattcttt
     7621 atttgaatta gaaaatggga gaaaaagaat gttggcttct gctggagaat tgcaaaaggg
     7681 taatgaatta gcattgcctt ccaaatatgt taacttcttg tatttagctt cacactatga
     7741 aaagttgaaa gggtcaccag aagataacga gcaaaaacaa ttatttgttg aacaacacaa
     7801 acactactta gatgagatta tagaacaaat tagtgaattc agtaaaagag tgatattagc
     7861 tgatgcaaat ttagataaag ttttgtcagc ctataacaaa catagagata agccaattag
     7921 agaacaagca gaaaacatta ttcacttatt tacccttacc aatttaggag cacctgctgc
     7981 tttcaagtat tttgatacaa caattgatcg taaaagatat acctcaacaa aagaagtctt
     8041 agacgccacc ttaattcatc aatcaatcac tggattgtat gagacaagaa ttgatttgtc
     8101 tcaattgggt ggtgatgaag gggctgatcc taagaagaaa agaaaagttg atccaaagaa
     8161 aaagcgtaag gtggatccta agaaaaagag aaaggttgac tacaaagacc atgacggtga
     8221 ttataaagat catgacatcg actacaagga tgacgatgac aagtgataat gactgcagag
     8281 atccatcgac ctgccgccaa gctaattccg ggcgaatttc tgtcgagtca tgtaattagt
     8341 tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt
     8401 tagacaacct gaagtctagg tccctattta tttttttata gttatgttag tattaagaac
     8461 gttatttata tttcaaattt ttcttttttt tcttggcggc gttagtatca aatcgacagc
     8521 agtatagcga ccagcattca catacgattg acgcatgata ttactttctg cgcacttaac
     8581 ttcgcatctg ggcagatgat gtcgaggcga aaaaaaatat aaatcacgct aacatttgat
     8641 taaaatagaa caactacaat ataaaaaaac tatacaaatg acaagttctt gaaaacaaga
     8701 atctttttat tgtcagtact gactcgagtt attatggaca tggcatagac atatacaaag
     8761 cttgttcacc atcggaagca gtaccatcgt ataaagcagt atccaaacca cacaaagtga
     8821 aacccattct tctataagca tgaatagctg gagcattaac attggtaact tccaaccaca
     8881 aatgaccagc acctctttct ctggcgaatt cagtagccaa acccatcaaa gctctaccaa
     8941 caccatgacc tctatgttct ggagcaactt caatatcttc aacagtcaat cttctgttcc
     9001 aaccagaata agaaacaaca acgaaaccag ccaaatcacc atcatcacca taagcaacga
     9061 aagttctaga atctggatca ccatcttcac cagcatcgga ttcatcatcg gattcatcat
     9121 ctgggaaaac cttagtcaat ggtggatcaa ctggaacttc tctcaaagtg aaaccatcac
     9181 cagtagcagt aactctaaaa acagtatcgg tagtgaaaga accatccaaa gcttcaatag
     9241 cttcagcatc acctggaaca gaagttctgt atctataagc agtatcatcc aaagtagtag
     9301 acataattgt aggatccggt tgtttatgtt cggatgtgat gtgagaactg tatcctagca
     9361 agattttaaa aggaagtata tgaaagaaga acctcagtgg caaatcctaa ccttttatat
     9421 ttctctacag gggcgcggcg tggggacaat tcaacgcgtc tgtgagggga gcgtttccct
     9481 gctcgcaggt ctgcagcgag gagccgtaat ttttgcttcg cgccgtgcgg ccatcaaaat
     9541 gtatggatgc aaatgattat acatggggat gtatgggcta aatgtacggg cgacagtcac
     9601 atcatgcccc tgagctgcgc acgtcaagac tgtcaaggag ggtattctgg gcctccatgt
     9661 cgctggccgg gtgacccggc ggggacgagg caagcttgat gcggccgctc tttgaaaaga
     9721 taatgtatga ttttgctttc actcatattt atacagaaac ttgatgtttt ctttcgagta
     9781 tatacaaggt gattacatgt acgtttgaag tacaactcta gattttgtag tgccctcttg
     9841 ggctagcggt aaaggtgcgc attttttcac accctacaat gttctgttca aaagattttg
     9901 gtcaaacgct gtagaagtga aagttggtgc gcatgtttcg gcgttcgaaa cttctccgca
     9961 gtgaaagata aatgatcgga gacggaattc cgtctcgttt tagagctaga aatagcaagt
    10021 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttc
    10081 tcgagttttt ttatcgagtg tttaaggata atgataactg aagagaagaa ttagttttgc
    10141 cgccaccgcg ggtttgcctc tgattaaata aaaaaaagct ggtgcttttt tttcttttat
    10201 aggaacatct tgaatatatg aactaattaa atgataattt tttacccatc tttactctta
    10261 atcactgagc tgcagtcaaa gaaaaaggga tacagcacct ggtgaagaga tgaacggaga
    10321 ctaacttaga cgcgttgatt cttttcaatt gcacatttta ttaatcgatg ctaacgtcta
    10381 tttacatata ttctttagag atattatcta gggcttcaaa taatctctgg acagcaataa
    10441 aagtctcttc aaaagtattg tataacggca atggggctaa tctgattaca tctggtcttc
    10501 tttcgtcaca gattatagca tgatcatgca agtacgcatt aactcgttcc atgacgttct
    10561 tgcccttttc atcgaaatgc ggttgaaaca taatggacaa ttgacatcct ctttcagctg
    10621 gattcaaagg agttaaaatt ttaaacccaa atttggagtt tgatgtactg gattgtggta
    10681 tgtaatactt ggaattcgtc aatagatcct gtaaaaattg agtcaaagca acactttttt
    10741 cacgaagttt agatactcca cccactttag catacacttc caatgacgac ttcacagcaa
    10801 caacatcaag aacagaagga tttgactgtc tgtaagaaag agccgagttt attggatcaa
    10861 actcttctaa cattttgaat cgttcttggg agttattgcc ccaccaacca gctagtctag
    10921 gaacgaaact gctcttcttg ttctctatag tgtatttttc atgcacaaaa attccaccta
    10981 tggctccagg tcccgagttt aaatatttgt aggaacacca agcagcaaaa tctactcccc
    11041 aatcatgtaa atttaatgga acattcccaa ctgcatgggc aagatcccac ccaactttaa
    11101 tttgttggct cttttcctta gcgtatttag ttatttcctc tatcttgaaa aattgaccag
    11161 tgtagtattg gataccagga aaacacacta gagccaattc atccaggttc tcatctatag
    11221 ccttgattat tctttctgtt ttaatataag tttcaccagg ttgaacttcc aattgaatca
    11281 aatgtttctc gtcgtatccg aacaatttaa caatgttcaa aaatgcatag tagtcagaag
    11341 gaaatgcttg tttttcaaat aaaattttgg ttcttttccc ctcaggtttg taaaaatgga
    11401 tcaacaatgc attcaagttt gctgttaaag aacccataac tgcaacttcg ttttcctttg
    11461 caccaacaat gggggctatt aatggtaata agggtaaatc gatgtctacc cacggtgtta
    11521 acagtttgtc aggatgattg aaatgagact caacccctcg ttcaacccat gcattcaatt
    11581 catcattgat agctttcttt gtattcttag gcatcaaccc aagagagttt ccacataaat
    11641 aaatagactc agttgatgac tcatatttat tatttttgat acctaatgat ccaaaagttg
    11701 gtatggcaaa ctcattttta aaagttggga actttttgtc caatttcttt gcctcggcta
    11761 atgacatctg ataataaaat ggggttggag tagttggtgg tataaccgga gagatagaat
    11821 tgaagaaaaa aatcggaaac aacaaaaaaa gttgataccc agtattatgt gggagataat
    11881 tgcgaatggt ggaaaaaaaa aagacgcctt tgggcctcaa caacaattct aacagcttga
    11941 gagctttaca atcgagaaac tatgattcat tccgttttaa tatgtatgta cttagtaaac
    12001 tcgtgaattt tatttgtggt ctactttagt actaacataa tcattggata gtcaataatg
    12061 atggtcttcc gagactaatg aaattctata ccaaagtcga tattccaaca cagaaattgc
    12121 tcttgcaaca agtgcacctg ttgatatcta ctagatcagc acaagagtgt tcggagctcc
    12181 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
    12241 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
    12301 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
    12361 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
    12421 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
    12481 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
    12541 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
    12601 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
    12661 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
    12721 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
    12781 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
    12841 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
    12901 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
    12961 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
    13021 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
    13081 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
    13141 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
    13201 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
    13261 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
    13321 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
    13381 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
    13441 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
    13501 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
    13561 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
    13621 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
    13681 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
    13741 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
    13801 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
    13861 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
    13921 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
    13981 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
    14041 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
    14101 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
    14161 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
    14221 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
    14281 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
    14341 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcctt ttcatcacgt
    14401 gctataaaaa taattataat ttaaattttt taatataaat atataaatta aaaatagaaa
    14461 gtaaaaaaag aaattaaaga aaaaatagtt tttgttttcc gaagatgtaa aagactctag
    14521 ggggatcgcc aacaaatact accttttatc ttgctcttcc tgctctcagg tattaatgcc
    14581 gaattgtttc atcttgtctg tgtagaagac cacacacgaa aatcctgtga ttttacattt
    14641 tacttatcgt taatcgaatg tatatctatt taatctgctt ttcttgtcta ataaatatat
    14701 atgtaaagta cgctttttgt tgaaattttt taaacctttg tttatttttt tttcttcatt
    14761 ccgtaactct tctaccttct ttatttactt tctaaaatcc aaatacaaaa cataaaaata
    14821 aataaacaca gagtaaattc ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa
    14881 gttacaggca agcgatccgt ctaagaaacc attattatca tgacattaac ctataaaaat
    14941 aggcgtatca cgaggccctt tcgtc
//
