LOCUS       sequence-212636-       14965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Solo vector pV1382 + sgScADE2.
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: pV1386
FEATURES             Location/Qualifiers
     source          1..14965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..45)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     148..167
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        191..406
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             407..1210
                     /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(1341..1796)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1478..1499)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1690..1709
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1922..1944
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1936..1953
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1937..1953
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1963..1982
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1963..1981
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3569..3981
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             4001..8104
                     /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             4001..8104
                     /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             8117..8137
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8141..8161
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8165..8185
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8186..8251
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(8332..8350)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      complement(8508..8705)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(9306..9649)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        9697..9965
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        9987..10062
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(12182..12202)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12182..12200)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12221..12237)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12221..12237)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12234..12256)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12245..12261
                     /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(12269..12275,12276..12293,12294..12299))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12314..12335
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12452..12469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12623..13211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12703..12722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13382..14173,14174..14242))
                     /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     14005..14024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14243..14347)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    14384..14887
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     14928..14946
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagatc tcggtcacag
       61 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
      121 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtaccg agagtgcacc
      181 ataccacctt ttcaattcat catttttttt ttattctttt ttttgatttc ggtttccttg
      241 aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact
      301 tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt
      361 aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt cgaaagctac
      421 atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat
      481 gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact
      541 ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt
      601 gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa
      661 ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca aattgcagta
      721 ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt
      781 gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag
      841 aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac
      901 taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca
      961 aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg
     1021 tttagatgac aagggtgatg cattgggtca acagtataga accgtggatg atgtggtctc
     1081 tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa
     1141 ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca
     1201 gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc
     1261 ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac agatgcgtaa
     1321 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     1381 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     1441 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     1501 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     1561 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     1621 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     1681 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     1741 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     1801 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     1861 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     1921 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag
     1981 ggcgaattgg gtacctgcca ctactaccac tgggagtttc gttcttctcg atactattag
     2041 ctttacttcc tgcactagca gtggttggat caacagaatc ttcataatca tcaaaatcgt
     2101 cttttgaaga ccccccgttt gatgtatggc cctgtctttt catcaaactt tttatatagt
     2161 tgactgaact gaggctaaat atgtgatcat cttcactata gacaatcttt ctcttatttg
     2221 caccaccgcc accaccagtc tttgagaaat tctcaaaacc ttttacgata ttaccaagcg
     2281 ggctctcttc gaaataatct atctcttttt gatatatcga atcctctagc gtggttagct
     2341 ttctagttag ttcttgcttc ttaagaattt gctggattag tttatttttc aattcaacgt
     2401 atttctcaga gtcatcttta gattttgatg aagatgtgcg ttcattcgct atatccttct
     2461 tggtcgtgtc ttttcgatcc tccttggctg gcactgaact cgtctttttt ggcgttgctg
     2521 ttccagacag acttatctca ttagatttgg aacttgtggg tttaacatca tttgcatctt
     2581 tagtagacat gattgtgtaa taccgtgatt gtttgttttg aaaggtctgt catatttcta
     2641 tcaatttcaa aacaaaatgt tcatcagaaa aaagccaaaa atgtctcttc tagcttctta
     2701 gtggtgtcgc aaaatacaca atgtcgctca acaatccaca ttcccggcgc atagctcaaa
     2761 tcacatgact acagctaaca attacacaaa aaaattctct tttcgatgta gcaactatct
     2821 tcaactaaaa cattttctcc ttcggcccat gattgtcctc cgggtcgaca gcaagccgtt
     2881 acaattgaga tggaaagcga cctaccttca ctcgataagg tgcttaattg tacttcatat
     2941 aaatctggcc cggatctaaa caaatgagtt ccattaagcc gtgggttctc aattagggtt
     3001 tttgtttttg atttagaaaa aagagatcaa gatttgttta caggtgatgc ctttttttag
     3061 aacttatgcg ttgcaaaagt tgactaacga tttctataag gtgatccaca ctaattatac
     3121 aaacgtacaa acagacatac ttttcctgcg ttcacctgat gttggccaga tttctctctt
     3181 cattgcatag aacataacca cactagggca acagaaaaaa aaaaaaaaag tgcatcggga
     3241 agttgtgttc cattcattat atgtctacta ctgcatatga gtagcccacc caccaccacc
     3301 atagtaagtt tttgtgtatg cgcgccgtca ggttatttca tttctgaatt tttcaaccac
     3361 cttactccct ttattgttga ttgacaattt tgctcacagt aagatctttt agactccaat
     3421 taatataaaa taagtctgat tttccaattc ctgttttttc tttttttttc tgtttctatt
     3481 tctttccttt tctccctttt ttttaattct tcattcaatc atcaattgat aattcaggaa
     3541 tattacaaca accatggccc ccacaagtga tcccccacac accatagctt caaaatgttt
     3601 ctactccttt tttactcttc cagattttct cggactccgc gcatcgccgt accacttcaa
     3661 aacacccaag cacagcatac taaatttccc ctctttcttc ctctagggtg tcgttaatta
     3721 cccgtactaa aggtttggaa aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa
     3781 aaaggcaata aaaattttta tcacgtttct ttttcttgaa aatttttttt ttgatttttt
     3841 tctctttcga tgacctccca ttgatattta agttaataaa cggtcttcaa tttctcaagt
     3901 ttcagtttca tttttcttgt tctattacaa ctttttttac ttcttgctca ttagaaagaa
     3961 agcatagcaa tctaatctaa gttttaatta caaacccggg atggataaaa agtatagtat
     4021 tggtttagat attggtacta actctgtggg ttgggcagtt atcactgacg aatataaagt
     4081 tccatcaaag aaatttaagg tgttaggtaa cactgacaga cactcaataa aaaagaatct
     4141 tatcggtgct cttttgttcg actccggtga aactgccgag gctacacgtt taaaaagaac
     4201 agcaagaaga agatataccc gtagaaaaaa tagaatatgt tatttacaag aaatcttttc
     4261 taatgaaatg gctaaagttg atgattcctt tttccataga ttggaagagt catttttggt
     4321 tgaagaagac aaaaagcatg agagacatcc aatctttggg aatatagttg atgaagtggc
     4381 ttaccatgaa aaatatccta ccatttatca tttaagaaag aaattggtag attcaactga
     4441 taaagctgac cttagattaa tctatttagc acttgcccat atgattaaat ttagaggtca
     4501 ttttttgatt gaaggtgatt tgaacccaga taattctgac gtggataaat tatttattca
     4561 attagtccaa acctacaacc aattatttga ggaaaatcca attaatgcta gtggtgtcga
     4621 tgccaaagct atattatcag ccagattatc aaaatctaga cgtttggaaa atttgattgc
     4681 ccaattgcca ggagaaaaaa agaatggatt atttggaaac ttgatcgcat tatcattggg
     4741 tttgacacca aattttaaat ctaattttga tttagctgaa gatgctaaat tacaattatc
     4801 aaaagacacc tatgacgacg atttggacaa tttacttgct caaattggtg atcaatatgc
     4861 agatttgttc ttagctgcta aaaacttatc tgatgctatt ttgttgtctg atattttgag
     4921 agtgaacaca gaaataacca aagctccatt atcagcatct atgatcaaac gttatgatga
     4981 acaccatcag gatttgactt tattgaaagc tttggtgaga caacaattgc cagagaagta
     5041 taaagaaatc tttttcgatc aatctaaaaa cgggtatgca ggttatattg atgggggtgc
     5101 ctcccaagag gaattttaca aatttataaa acctatttta gaaaagatgg atgggactga
     5161 ggaacttttg gtcaaattga acagagaaga tttgttacgt aaacagagaa cttttgataa
     5221 tggtagtata cctcaccaaa ttcatttggg tgagttgcat gcaattttaa gaagacaaga
     5281 agatttttat ccatttttaa aagataatag agaaaaaatc gagaaaattt taacctttag
     5341 aattccatac tatgttgggc ctttggctag aggtaattca agatttgcct ggatgacacg
     5401 taaatcagaa gaaactatta ccccttggaa ttttgaagag gttgttgata aaggagcatc
     5461 agcacagagt tttattgaaa gaatgaccaa tttcgataaa aacttaccaa atgaaaaagt
     5521 tttaccaaaa cattccttgt tatacgaata ttttactgtt tacaatgaac ttacaaaggt
     5581 taaatatgtt actgaaggta tgcgtaagcc agccttttta tctggagaac agaaaaaggc
     5641 aatagttgat ttattgttta aaacaaatag aaaagttact gttaaacaat taaaagaaga
     5701 ttactttaag aaaattgaat gttttgattc agttgaaatc agtggtgttg aagacagatt
     5761 taatgctagt ttaggaactt accatgattt acttaaaatt atcaaagata aagatttctt
     5821 ggataacgaa gaaaatgaag acattttaga agacattgtt ttaaccttaa ctttattcga
     5881 agatagagag atgattgaag aacgtttgaa gacttatgca catttgtttg acgataaagt
     5941 gatgaaacag ttgaaaagaa gacgttatac tggatggggt agattgtctc gtaaattgat
     6001 caatggaatt agagataaac aaagtggtaa aactatcttg gactttttga aatctgacgg
     6061 atttgctaat agaaatttca tgcaattgat ccacgacgat agtttgacat ttaaagaaga
     6121 catccaaaag gcccaagtga gtgggcaagg tgattcatta catgaacata ttgcaaattt
     6181 agccggatct cctgctatta agaaagggat attacaaact gttaaagttg tggatgaatt
     6241 agtgaaagta atgggaagac ataaacctga aaacattgtc attgagatgg caagagaaaa
     6301 tcaaactaca caaaaaggac agaaaaatag tagagaacgt atgaaaagaa tagaagaggg
     6361 tattaaagaa ttgggtagtc aaatattgaa agaacaccca gtggaaaata cccagttgca
     6421 aaatgaaaaa ttatatcttt actaccttca aaatggacgt gatatgtatg ttgatcagga
     6481 attagatata aatagacttt cagattatga tgtagatcat atagttccac aatctttctt
     6541 gaaagatgat tccatagaca ataaagtatt aactagaagt gataaaaata gaggtaaaag
     6601 tgataatgtc ccaagtgagg aagtcgtcaa aaagatgaaa aattactggc gtcaactttt
     6661 gaatgctaaa ttaattactc aaagaaaatt tgataatttg actaaagcag aaagaggtgg
     6721 gctttctgaa ttagataaag ccgggttcat taaaagacaa ttggtcgaaa ctagacaaat
     6781 tactaaacat gttgcccaaa ttttagattc ccgtatgaac actaagtatg acgaaaatga
     6841 taagttaata cgtgaggtta aagtcattac tttaaaatca aaacttgtct ctgatttcag
     6901 aaaggatttc caattctata aagttagaga aattaataat tatcatcatg ctcatgatgc
     6961 atatttgaat gctgtagttg gaactgcttt aatcaagaaa taccctaaat tagaatctga
     7021 atttgtatat ggtgattaca aagtctatga tgttagaaag atgattgcta aatcagaaca
     7081 agaaattggt aaagctacag ctaaatactt cttttactct aacattatga atttctttaa
     7141 aacagaaatt actttggcaa acggtgaaat tagaaaaaga cctcttattg aaacaaatgg
     7201 tgagactgga gagatagttt gggacaaagg gcgtgatttc gctactgtta gaaaagtttt
     7261 atcaatgcca caagttaaca ttgtaaagaa aacagaggtt caaactggtg gtttctcaaa
     7321 agaaagtatt ttgcctaaaa gaaatagtga taaattgatt gccagaaaaa aggattggga
     7381 tccaaagaaa tatggtggtt tcgactcacc aaccgtagcc tattctgttt tggttgtggc
     7441 aaaggttgaa aagggtaaaa gtaaaaagct taaatcagta aaagaacttt tgggtattac
     7501 aataatggaa agaagttcct ttgaaaagaa ccctattgat tttttggaag ctaaaggtta
     7561 taaggaagta aagaaggact taataatcaa attgcctaaa tattctttat ttgaattaga
     7621 aaatgggaga aaaagaatgt tggcttctgc tggagaattg caaaagggta atgaattagc
     7681 attgccttcc aaatatgtta acttcttgta tttagcttca cactatgaaa agttgaaagg
     7741 gtcaccagaa gataacgagc aaaaacaatt atttgttgaa caacacaaac actacttaga
     7801 tgagattata gaacaaatta gtgaattcag taaaagagtg atattagctg atgcaaattt
     7861 agataaagtt ttgtcagcct ataacaaaca tagagataag ccaattagag aacaagcaga
     7921 aaacattatt cacttattta cccttaccaa tttaggagca cctgctgctt tcaagtattt
     7981 tgatacaaca attgatcgta aaagatatac ctcaacaaaa gaagtcttag acgccacctt
     8041 aattcatcaa tcaatcactg gattgtatga gacaagaatt gatttgtctc aattgggtgg
     8101 tgatgaaggg gctgatccta agaagaaaag aaaagttgat ccaaagaaaa agcgtaaggt
     8161 ggatcctaag aaaaagagaa aggttgacta caaagaccat gacggtgatt ataaagatca
     8221 tgacatcgac tacaaggatg acgatgacaa gtgataatga ctgcagagat ccatcgacct
     8281 gccgccaagc taattccggg cgaatttctg tcgagtcatg taattagtta tgtcacgctt
     8341 acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     8401 agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt
     8461 tcaaattttt cttttttttc ttggcggcgt tagtatcaaa tcgacagcag tatagcgacc
     8521 agcattcaca tacgattgac gcatgatatt actttctgcg cacttaactt cgcatctggg
     8581 cagatgatgt cgaggcgaaa aaaaatataa atcacgctaa catttgatta aaatagaaca
     8641 actacaatat aaaaaaacta tacaaatgac aagttcttga aaacaagaat ctttttattg
     8701 tcagtactga ctcgagttat tatggacatg gcatagacat atacaaagct tgttcaccat
     8761 cggaagcagt accatcgtat aaagcagtat ccaaaccaca caaagtgaaa cccattcttc
     8821 tataagcatg aatagctgga gcattaacat tggtaacttc caaccacaaa tgaccagcac
     8881 ctctttctct ggcgaattca gtagccaaac ccatcaaagc tctaccaaca ccatgacctc
     8941 tatgttctgg agcaacttca atatcttcaa cagtcaatct tctgttccaa ccagaataag
     9001 aaacaacaac gaaaccagcc aaatcaccat catcaccata agcaacgaaa gttctagaat
     9061 ctggatcacc atcttcacca gcatcggatt catcatcgga ttcatcatct gggaaaacct
     9121 tagtcaatgg tggatcaact ggaacttctc tcaaagtgaa accatcacca gtagcagtaa
     9181 ctctaaaaac agtatcggta gtgaaagaac catccaaagc ttcaatagct tcagcatcac
     9241 ctggaacaga agttctgtat ctataagcag tatcatccaa agtagtagac ataattgtag
     9301 gatccggttg tttatgttcg gatgtgatgt gagaactgta tcctagcaag attttaaaag
     9361 gaagtatatg aaagaagaac ctcagtggca aatcctaacc ttttatattt ctctacaggg
     9421 gcgcggcgtg gggacaattc aacgcgtctg tgaggggagc gtttccctgc tcgcaggtct
     9481 gcagcgagga gccgtaattt ttgcttcgcg ccgtgcggcc atcaaaatgt atggatgcaa
     9541 atgattatac atggggatgt atgggctaaa tgtacgggcg acagtcacat catgcccctg
     9601 agctgcgcac gtcaagactg tcaaggaggg tattctgggc ctccatgtcg ctggccgggt
     9661 gacccggcgg ggacgaggca agcttgatgc ggccgctctt tgaaaagata atgtatgatt
     9721 ttgctttcac tcatatttat acagaaactt gatgttttct ttcgagtata tacaaggtga
     9781 ttacatgtac gtttgaagta caactctaga ttttgtagtg ccctcttggg ctagcggtaa
     9841 aggtgcgcat tttttcacac cctacaatgt tctgttcaaa agattttggt caaacgctgt
     9901 agaagtgaaa gttggtgcgc atgtttcggc gttcgaaact tctccgcagt gaaagataaa
     9961 tgatcgattg ggacgtatga ttgttggttt tagagctaga aatagcaagt taaaataagg
    10021 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttc tcgagttttt
    10081 ttatcgagtg tttaaggata atgataactg aagagaagaa ttagttttgc cgccaccgcg
    10141 ggtttgcctc tgattaaata aaaaaaagct ggtgcttttt tttcttttat aggaacatct
    10201 tgaatatatg aactaattaa atgataattt tttacccatc tttactctta atcactgagc
    10261 tgcagtcaaa gaaaaaggga tacagcacct ggtgaagaga tgaacggaga ctaacttaga
    10321 cgcgttgatt cttttcaatt gcacatttta ttaatcgatg ctaacgtcta tttacatata
    10381 ttctttagag atattatcta gggcttcaaa taatctctgg acagcaataa aagtctcttc
    10441 aaaagtattg tataacggca atggggctaa tctgattaca tctggtcttc tttcgtcaca
    10501 gattatagca tgatcatgca agtacgcatt aactcgttcc atgacgttct tgcccttttc
    10561 atcgaaatgc ggttgaaaca taatggacaa ttgacatcct ctttcagctg gattcaaagg
    10621 agttaaaatt ttaaacccaa atttggagtt tgatgtactg gattgtggta tgtaatactt
    10681 ggaattcgtc aatagatcct gtaaaaattg agtcaaagca acactttttt cacgaagttt
    10741 agatactcca cccactttag catacacttc caatgacgac ttcacagcaa caacatcaag
    10801 aacagaagga tttgactgtc tgtaagaaag agccgagttt attggatcaa actcttctaa
    10861 cattttgaat cgttcttggg agttattgcc ccaccaacca gctagtctag gaacgaaact
    10921 gctcttcttg ttctctatag tgtatttttc atgcacaaaa attccaccta tggctccagg
    10981 tcccgagttt aaatatttgt aggaacacca agcagcaaaa tctactcccc aatcatgtaa
    11041 atttaatgga acattcccaa ctgcatgggc aagatcccac ccaactttaa tttgttggct
    11101 cttttcctta gcgtatttag ttatttcctc tatcttgaaa aattgaccag tgtagtattg
    11161 gataccagga aaacacacta gagccaattc atccaggttc tcatctatag ccttgattat
    11221 tctttctgtt ttaatataag tttcaccagg ttgaacttcc aattgaatca aatgtttctc
    11281 gtcgtatccg aacaatttaa caatgttcaa aaatgcatag tagtcagaag gaaatgcttg
    11341 tttttcaaat aaaattttgg ttcttttccc ctcaggtttg taaaaatgga tcaacaatgc
    11401 attcaagttt gctgttaaag aacccataac tgcaacttcg ttttcctttg caccaacaat
    11461 gggggctatt aatggtaata agggtaaatc gatgtctacc cacggtgtta acagtttgtc
    11521 aggatgattg aaatgagact caacccctcg ttcaacccat gcattcaatt catcattgat
    11581 agctttcttt gtattcttag gcatcaaccc aagagagttt ccacataaat aaatagactc
    11641 agttgatgac tcatatttat tatttttgat acctaatgat ccaaaagttg gtatggcaaa
    11701 ctcattttta aaagttggga actttttgtc caatttcttt gcctcggcta atgacatctg
    11761 ataataaaat ggggttggag tagttggtgg tataaccgga gagatagaat tgaagaaaaa
    11821 aatcggaaac aacaaaaaaa gttgataccc agtattatgt gggagataat tgcgaatggt
    11881 ggaaaaaaaa aagacgcctt tgggcctcaa caacaattct aacagcttga gagctttaca
    11941 atcgagaaac tatgattcat tccgttttaa tatgtatgta cttagtaaac tcgtgaattt
    12001 tatttgtggt ctactttagt actaacataa tcattggata gtcaataatg atggtcttcc
    12061 gagactaatg aaattctata ccaaagtcga tattccaaca cagaaattgc tcttgcaaca
    12121 agtgcacctg ttgatatcta ctagatcagc acaagagtgt tcggagctcc agcttttgtt
    12181 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
    12241 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
    12301 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
    12361 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
    12421 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
    12481 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
    12541 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
    12601 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
    12661 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
    12721 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
    12781 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
    12841 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
    12901 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
    12961 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
    13021 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
    13081 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
    13141 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
    13201 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
    13261 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
    13321 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
    13381 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
    13441 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
    13501 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
    13561 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
    13621 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
    13681 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
    13741 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
    13801 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
    13861 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
    13921 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
    13981 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
    14041 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
    14101 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
    14161 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
    14221 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
    14281 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
    14341 ttccgcgcac atttccccga aaagtgccac ctgggtcctt ttcatcacgt gctataaaaa
    14401 taattataat ttaaattttt taatataaat atataaatta aaaatagaaa gtaaaaaaag
    14461 aaattaaaga aaaaatagtt tttgttttcc gaagatgtaa aagactctag ggggatcgcc
    14521 aacaaatact accttttatc ttgctcttcc tgctctcagg tattaatgcc gaattgtttc
    14581 atcttgtctg tgtagaagac cacacacgaa aatcctgtga ttttacattt tacttatcgt
    14641 taatcgaatg tatatctatt taatctgctt ttcttgtcta ataaatatat atgtaaagta
    14701 cgctttttgt tgaaattttt taaacctttg tttatttttt tttcttcatt ccgtaactct
    14761 tctaccttct ttatttactt tctaaaatcc aaatacaaaa cataaaaata aataaacaca
    14821 gagtaaattc ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa gttacaggca
    14881 agcgatccgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
    14941 cgaggccctt tcgtcgaatt cgcgc
//
