LOCUS       sequence-212601-       14519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Solo vector pV1326 + sgScADE2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14519)
  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 14519)
  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: pV1338
FEATURES             Location/Qualifiers
     source          1..14519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(32..54)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     157..176
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        200..415
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             416..1219
                     /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(1350..1805)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1487..1508)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1699..1718
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1931..1953
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1945..1962
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1946..1962
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1972..1991
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1972..1990
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3564..7667
                     /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             3564..7667
                     /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             7680..7700
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7704..7724
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7728..7748
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7749..7814
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(7895..7913)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      complement(8071..8268)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(8869..9212)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        9260..9528
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        9550..9625
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(11745..11765)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11745..11763)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11784..11800)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11784..11800)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11797..11819)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11808..11824
                     /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(11832..11838,11839..11856,11857..11862))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11877..11898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12015..12032)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12186..12774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12266..12285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12945..13736,13737..13805))
                     /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     13568..13587
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13806..13910)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    13947..14450
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     14491..14509
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aattcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagatct
       61 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
      121 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtaccga
      181 gagtgcacca taccaccttt tcaattcatc attttttttt tattcttttt tttgatttcg
      241 gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga
      301 gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc
      361 agtattctta acccaactgc acagaacaaa aacctgcagg aaacgaagat aaatcatgtc
      421 gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt
      481 taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa
      541 ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt
      601 ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc
      661 caagtacaat tttttactct tcgaagacag aaaatttgct gacattggta atacagtcaa
      721 attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca
      781 cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa
      841 ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg
      901 agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt
      961 tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc
     1021 cggtgtgggt ttagatgaca agggtgatgc attgggtcaa cagtatagaa ccgtggatga
     1081 tgtggtctct acaggatctg acattattat tgttggaaga ggactatttg caaagggaag
     1141 ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag
     1201 atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca
     1261 aattagagct tcaatttaat tatatcagtt attaccctat gcggtgtgaa ataccgcaca
     1321 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
     1381 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     1441 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     1501 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     1561 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     1621 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     1681 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     1741 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     1801 cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     1861 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     1921 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac
     1981 tcactatagg gcgaattggg tacctgccac tactaccact gggagtttcg ttcttctcga
     2041 tactattagc tttacttcct gcactagcag tggttggatc aacagaatct tcataatcat
     2101 caaaatcgtc ttttgaagac cccccgtttg atgtatggcc ctgtcttttc atcaaacttt
     2161 ttatatagtt gactgaactg aggctaaata tgtgatcatc ttcactatag acaatctttc
     2221 tcttatttgc accaccgcca ccaccagtct ttgagaaatt ctcaaaacct tttacgatat
     2281 taccaagcgg gctctcttcg aaataatcta tctctttttg atatatcgaa tcctctagcg
     2341 tggttagctt tctagttagt tcttgcttct taagaatttg ctggattagt ttatttttca
     2401 attcaacgta tttctcagag tcatctttag attttgatga agatgtgcgt tcattcgcta
     2461 tatccttctt ggtcgtgtct tttcgatcct ccttggctgg cactgaactc gtcttttttg
     2521 gcgttgctgt tccagacaga cttatctcat tagatttgga acttgtgggt ttaacatcat
     2581 ttgcatcttt agtagacatg attgtgtaat accgtgattg tttgttttga aaggtctgtc
     2641 atatttctat caatttcaaa acaaaatgtt catcagaaaa aagccaaaaa tgtctcttct
     2701 agcttcttag tggtgtcgca aaatacacaa tgtcgctcaa caatccacat tcccggcgca
     2761 tagctcaaat cacatgacta cagctaacaa ttacacaaaa aaattctctt ttcgatgtag
     2821 caactatctt caactaaaac attttctcct tcggcccatg attgtcctcc gggtcgacag
     2881 caagccgtta caattgagat ggaaagcgac ctaccttcac tcgataaggt gcttaattgt
     2941 acttcatata aatctggccc ggatctaaac aaatgagttc cattaagccg tgggttctca
     3001 attagggttt ttgtttttga tttagaaaaa agagatcaag atttgtttac aggtgatgcc
     3061 tttttttaga acttatgcgt tgcaaaagtt gactaacgat ttctataagg tgatccacac
     3121 taattataca aacgtacaaa cagacatact tttcctgcgt tcacctgatg ttggccagat
     3181 ttctctcttc attgcataga acataaccac actagggcaa cagaaaaaaa aaaaaaaagt
     3241 gcatcgggaa gttgtgttcc attcattata tgtctactac tgcatatgag tagcccaccc
     3301 accaccacca tagtaagttt ttgtgtatgc gcgccgtcag gttatttcat ttctgaattt
     3361 ttcaaccacc ttactccctt tattgttgat tgacaatttt gctcacagta agatctttta
     3421 gactccaatt aatataaaat aagtctgatt ttccaattcc tgttttttct ttttttttct
     3481 gtttctattt ctttcctttt ctcccttttt tttaattctt cattcaatca tcaattgata
     3541 attcaggaat attacaaccc gggatggata aaaagtatag tattggttta gatattggta
     3601 ctaactctgt gggttgggca gttatcactg acgaatataa agttccatca aagaaattta
     3661 aggtgttagg taacactgac agacactcaa taaaaaagaa tcttatcggt gctcttttgt
     3721 tcgactccgg tgaaactgcc gaggctacac gtttaaaaag aacagcaaga agaagatata
     3781 cccgtagaaa aaatagaata tgttatttac aagaaatctt ttctaatgaa atggctaaag
     3841 ttgatgattc ctttttccat agattggaag agtcattttt ggttgaagaa gacaaaaagc
     3901 atgagagaca tccaatcttt gggaatatag ttgatgaagt ggcttaccat gaaaaatatc
     3961 ctaccattta tcatttaaga aagaaattgg tagattcaac tgataaagct gaccttagat
     4021 taatctattt agcacttgcc catatgatta aatttagagg tcattttttg attgaaggtg
     4081 atttgaaccc agataattct gacgtggata aattatttat tcaattagtc caaacctaca
     4141 accaattatt tgaggaaaat ccaattaatg ctagtggtgt cgatgccaaa gctatattat
     4201 cagccagatt atcaaaatct agacgtttgg aaaatttgat tgcccaattg ccaggagaaa
     4261 aaaagaatgg attatttgga aacttgatcg cattatcatt gggtttgaca ccaaatttta
     4321 aatctaattt tgatttagct gaagatgcta aattacaatt atcaaaagac acctatgacg
     4381 acgatttgga caatttactt gctcaaattg gtgatcaata tgcagatttg ttcttagctg
     4441 ctaaaaactt atctgatgct attttgttgt ctgatatttt gagagtgaac acagaaataa
     4501 ccaaagctcc attatcagca tctatgatca aacgttatga tgaacaccat caggatttga
     4561 ctttattgaa agctttggtg agacaacaat tgccagagaa gtataaagaa atctttttcg
     4621 atcaatctaa aaacgggtat gcaggttata ttgatggggg tgcctcccaa gaggaatttt
     4681 acaaatttat aaaacctatt ttagaaaaga tggatgggac tgaggaactt ttggtcaaat
     4741 tgaacagaga agatttgtta cgtaaacaga gaacttttga taatggtagt atacctcacc
     4801 aaattcattt gggtgagttg catgcaattt taagaagaca agaagatttt tatccatttt
     4861 taaaagataa tagagaaaaa atcgagaaaa ttttaacctt tagaattcca tactatgttg
     4921 ggcctttggc tagaggtaat tcaagatttg cctggatgac acgtaaatca gaagaaacta
     4981 ttaccccttg gaattttgaa gaggttgttg ataaaggagc atcagcacag agttttattg
     5041 aaagaatgac caatttcgat aaaaacttac caaatgaaaa agttttacca aaacattcct
     5101 tgttatacga atattttact gtttacaatg aacttacaaa ggttaaatat gttactgaag
     5161 gtatgcgtaa gccagccttt ttatctggag aacagaaaaa ggcaatagtt gatttattgt
     5221 ttaaaacaaa tagaaaagtt actgttaaac aattaaaaga agattacttt aagaaaattg
     5281 aatgttttga ttcagttgaa atcagtggtg ttgaagacag atttaatgct agtttaggaa
     5341 cttaccatga tttacttaaa attatcaaag ataaagattt cttggataac gaagaaaatg
     5401 aagacatttt agaagacatt gttttaacct taactttatt cgaagataga gagatgattg
     5461 aagaacgttt gaagacttat gcacatttgt ttgacgataa agtgatgaaa cagttgaaaa
     5521 gaagacgtta tactggatgg ggtagattgt ctcgtaaatt gatcaatgga attagagata
     5581 aacaaagtgg taaaactatc ttggactttt tgaaatctga cggatttgct aatagaaatt
     5641 tcatgcaatt gatccacgac gatagtttga catttaaaga agacatccaa aaggcccaag
     5701 tgagtgggca aggtgattca ttacatgaac atattgcaaa tttagccgga tctcctgcta
     5761 ttaagaaagg gatattacaa actgttaaag ttgtggatga attagtgaaa gtaatgggaa
     5821 gacataaacc tgaaaacatt gtcattgaga tggcaagaga aaatcaaact acacaaaaag
     5881 gacagaaaaa tagtagagaa cgtatgaaaa gaatagaaga gggtattaaa gaattgggta
     5941 gtcaaatatt gaaagaacac ccagtggaaa atacccagtt gcaaaatgaa aaattatatc
     6001 tttactacct tcaaaatgga cgtgatatgt atgttgatca ggaattagat ataaatagac
     6061 tttcagatta tgatgtagat catatagttc cacaatcttt cttgaaagat gattccatag
     6121 acaataaagt attaactaga agtgataaaa atagaggtaa aagtgataat gtcccaagtg
     6181 aggaagtcgt caaaaagatg aaaaattact ggcgtcaact tttgaatgct aaattaatta
     6241 ctcaaagaaa atttgataat ttgactaaag cagaaagagg tgggctttct gaattagata
     6301 aagccgggtt cattaaaaga caattggtcg aaactagaca aattactaaa catgttgccc
     6361 aaattttaga ttcccgtatg aacactaagt atgacgaaaa tgataagtta atacgtgagg
     6421 ttaaagtcat tactttaaaa tcaaaacttg tctctgattt cagaaaggat ttccaattct
     6481 ataaagttag agaaattaat aattatcatc atgctcatga tgcatatttg aatgctgtag
     6541 ttggaactgc tttaatcaag aaatacccta aattagaatc tgaatttgta tatggtgatt
     6601 acaaagtcta tgatgttaga aagatgattg ctaaatcaga acaagaaatt ggtaaagcta
     6661 cagctaaata cttcttttac tctaacatta tgaatttctt taaaacagaa attactttgg
     6721 caaacggtga aattagaaaa agacctctta ttgaaacaaa tggtgagact ggagagatag
     6781 tttgggacaa agggcgtgat ttcgctactg ttagaaaagt tttatcaatg ccacaagtta
     6841 acattgtaaa gaaaacagag gttcaaactg gtggtttctc aaaagaaagt attttgccta
     6901 aaagaaatag tgataaattg attgccagaa aaaaggattg ggatccaaag aaatatggtg
     6961 gtttcgactc accaaccgta gcctattctg ttttggttgt ggcaaaggtt gaaaagggta
     7021 aaagtaaaaa gcttaaatca gtaaaagaac ttttgggtat tacaataatg gaaagaagtt
     7081 cctttgaaaa gaaccctatt gattttttgg aagctaaagg ttataaggaa gtaaagaagg
     7141 acttaataat caaattgcct aaatattctt tatttgaatt agaaaatggg agaaaaagaa
     7201 tgttggcttc tgctggagaa ttgcaaaagg gtaatgaatt agcattgcct tccaaatatg
     7261 ttaacttctt gtatttagct tcacactatg aaaagttgaa agggtcacca gaagataacg
     7321 agcaaaaaca attatttgtt gaacaacaca aacactactt agatgagatt atagaacaaa
     7381 ttagtgaatt cagtaaaaga gtgatattag ctgatgcaaa tttagataaa gttttgtcag
     7441 cctataacaa acatagagat aagccaatta gagaacaagc agaaaacatt attcacttat
     7501 ttacccttac caatttagga gcacctgctg ctttcaagta ttttgataca acaattgatc
     7561 gtaaaagata tacctcaaca aaagaagtct tagacgccac cttaattcat caatcaatca
     7621 ctggattgta tgagacaaga attgatttgt ctcaattggg tggtgatgaa ggggctgatc
     7681 ctaagaagaa aagaaaagtt gatccaaaga aaaagcgtaa ggtggatcct aagaaaaaga
     7741 gaaaggttga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     7801 atgacgatga caagtgataa tgactgcaga gatccatcga cctgccgcca agctaattcc
     7861 gggcgaattt ctgtcgagtc atgtaattag ttatgtcacg cttacattca cgccctcccc
     7921 ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt
     7981 atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     8041 ttcttggcgg cgttagtatc aaatcgacag cagtatagcg accagcattc acatacgatt
     8101 gacgcatgat attactttct gcgcacttaa cttcgcatct gggcagatga tgtcgaggcg
     8161 aaaaaaaata taaatcacgc taacatttga ttaaaataga acaactacaa tataaaaaaa
     8221 ctatacaaat gacaagttct tgaaaacaag aatcttttta ttgtcagtac tgactcgagt
     8281 tattatggac atggcataga catatacaaa gcttgttcac catcggaagc agtaccatcg
     8341 tataaagcag tatccaaacc acacaaagtg aaacccattc ttctataagc atgaatagct
     8401 ggagcattaa cattggtaac ttccaaccac aaatgaccag cacctctttc tctggcgaat
     8461 tcagtagcca aacccatcaa agctctacca acaccatgac ctctatgttc tggagcaact
     8521 tcaatatctt caacagtcaa tcttctgttc caaccagaat aagaaacaac aacgaaacca
     8581 gccaaatcac catcatcacc ataagcaacg aaagttctag aatctggatc accatcttca
     8641 ccagcatcgg attcatcatc ggattcatca tctgggaaaa ccttagtcaa tggtggatca
     8701 actggaactt ctctcaaagt gaaaccatca ccagtagcag taactctaaa aacagtatcg
     8761 gtagtgaaag aaccatccaa agcttcaata gcttcagcat cacctggaac agaagttctg
     8821 tatctataag cagtatcatc caaagtagta gacataattg taggatccgg ttgtttatgt
     8881 tcggatgtga tgtgagaact gtatcctagc aagattttaa aaggaagtat atgaaagaag
     8941 aacctcagtg gcaaatccta accttttata tttctctaca ggggcgcggc gtggggacaa
     9001 ttcaacgcgt ctgtgagggg agcgtttccc tgctcgcagg tctgcagcga ggagccgtaa
     9061 tttttgcttc gcgccgtgcg gccatcaaaa tgtatggatg caaatgatta tacatgggga
     9121 tgtatgggct aaatgtacgg gcgacagtca catcatgccc ctgagctgcg cacgtcaaga
     9181 ctgtcaagga gggtattctg ggcctccatg tcgctggccg ggtgacccgg cggggacgag
     9241 gcaagcttga tgcggccgct ctttgaaaag ataatgtatg attttgcttt cactcatatt
     9301 tatacagaaa cttgatgttt tctttcgagt atatacaagg tgattacatg tacgtttgaa
     9361 gtacaactct agattttgta gtgccctctt gggctagcgg taaaggtgcg cattttttca
     9421 caccctacaa tgttctgttc aaaagatttt ggtcaaacgc tgtagaagtg aaagttggtg
     9481 cgcatgtttc ggcgttcgaa acttctccgc agtgaaagat aaatgatcga ttgggacgta
     9541 tgattgttgg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     9601 tgaaaaagtg gcaccgagtc ggtgcttttt ttctcgagtt tttttatcga gtgtttaagg
     9661 ataatgataa ctgaagagaa gaattagttt tgccgccacc gcgggtttgc ctctgattaa
     9721 ataaaaaaaa gctggtgctt ttttttcttt tataggaaca tcttgaatat atgaactaat
     9781 taaatgataa ttttttaccc atctttactc ttaatcactg agctgcagtc aaagaaaaag
     9841 ggatacagca cctggtgaag agatgaacgg agactaactt agacgcgttg attcttttca
     9901 attgcacatt ttattaatcg atgctaacgt ctatttacat atattcttta gagatattat
     9961 ctagggcttc aaataatctc tggacagcaa taaaagtctc ttcaaaagta ttgtataacg
    10021 gcaatggggc taatctgatt acatctggtc ttctttcgtc acagattata gcatgatcat
    10081 gcaagtacgc attaactcgt tccatgacgt tcttgccctt ttcatcgaaa tgcggttgaa
    10141 acataatgga caattgacat cctctttcag ctggattcaa aggagttaaa attttaaacc
    10201 caaatttgga gtttgatgta ctggattgtg gtatgtaata cttggaattc gtcaatagat
    10261 cctgtaaaaa ttgagtcaaa gcaacacttt tttcacgaag tttagatact ccacccactt
    10321 tagcatacac ttccaatgac gacttcacag caacaacatc aagaacagaa ggatttgact
    10381 gtctgtaaga aagagccgag tttattggat caaactcttc taacattttg aatcgttctt
    10441 gggagttatt gccccaccaa ccagctagtc taggaacgaa actgctcttc ttgttctcta
    10501 tagtgtattt ttcatgcaca aaaattccac ctatggctcc aggtcccgag tttaaatatt
    10561 tgtaggaaca ccaagcagca aaatctactc cccaatcatg taaatttaat ggaacattcc
    10621 caactgcatg ggcaagatcc cacccaactt taatttgttg gctcttttcc ttagcgtatt
    10681 tagttatttc ctctatcttg aaaaattgac cagtgtagta ttggatacca ggaaaacaca
    10741 ctagagccaa ttcatccagg ttctcatcta tagccttgat tattctttct gttttaatat
    10801 aagtttcacc aggttgaact tccaattgaa tcaaatgttt ctcgtcgtat ccgaacaatt
    10861 taacaatgtt caaaaatgca tagtagtcag aaggaaatgc ttgtttttca aataaaattt
    10921 tggttctttt cccctcaggt ttgtaaaaat ggatcaacaa tgcattcaag tttgctgtta
    10981 aagaacccat aactgcaact tcgttttcct ttgcaccaac aatgggggct attaatggta
    11041 ataagggtaa atcgatgtct acccacggtg ttaacagttt gtcaggatga ttgaaatgag
    11101 actcaacccc tcgttcaacc catgcattca attcatcatt gatagctttc tttgtattct
    11161 taggcatcaa cccaagagag tttccacata aataaataga ctcagttgat gactcatatt
    11221 tattattttt gatacctaat gatccaaaag ttggtatggc aaactcattt ttaaaagttg
    11281 ggaacttttt gtccaatttc tttgcctcgg ctaatgacat ctgataataa aatggggttg
    11341 gagtagttgg tggtataacc ggagagatag aattgaagaa aaaaatcgga aacaacaaaa
    11401 aaagttgata cccagtatta tgtgggagat aattgcgaat ggtggaaaaa aaaaagacgc
    11461 ctttgggcct caacaacaat tctaacagct tgagagcttt acaatcgaga aactatgatt
    11521 cattccgttt taatatgtat gtacttagta aactcgtgaa ttttatttgt ggtctacttt
    11581 agtactaaca taatcattgg atagtcaata atgatggtct tccgagacta atgaaattct
    11641 ataccaaagt cgatattcca acacagaaat tgctcttgca acaagtgcac ctgttgatat
    11701 ctactagatc agcacaagag tgttcggagc tccagctttt gttcccttta gtgagggtta
    11761 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
    11821 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
    11881 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
    11941 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
    12001 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
    12061 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
    12121 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
    12181 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
    12241 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
    12301 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
    12361 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
    12421 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
    12481 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
    12541 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
    12601 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
    12661 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
    12721 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
    12781 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
    12841 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
    12901 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
    12961 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
    13021 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
    13081 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
    13141 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
    13201 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
    13261 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
    13321 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
    13381 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
    13441 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
    13501 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
    13561 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
    13621 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
    13681 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
    13741 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
    13801 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
    13861 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
    13921 cgaaaagtgc cacctgggtc cttttcatca cgtgctataa aaataattat aatttaaatt
    13981 ttttaatata aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa agaaaaaata
    14041 gtttttgttt tccgaagatg taaaagactc tagggggatc gccaacaaat actacctttt
    14101 atcttgctct tcctgctctc aggtattaat gccgaattgt ttcatcttgt ctgtgtagaa
    14161 gaccacacac gaaaatcctg tgattttaca ttttacttat cgttaatcga atgtatatct
    14221 atttaatctg cttttcttgt ctaataaata tatatgtaaa gtacgctttt tgttgaaatt
    14281 ttttaaacct ttgtttattt ttttttcttc attccgtaac tcttctacct tctttattta
    14341 ctttctaaaa tccaaataca aaacataaaa ataaataaac acagagtaaa ttcccaaatt
    14401 attccatcat taaaagatac gaggcgcgtg taagttacag gcaagcgatc cgtctaagaa
    14461 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtcg
//
