pV1326

LOCUS        pV1326             14517 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  S. cerevisiae and C. glabrata Solo CRISPR vector, marked with URA3 
            and NatR.
ACCESSION   .
VERSION     .
KEYWORDS    pV1326
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14517)
  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 14517)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14517
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     CDS             3565..7668
                     /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             7681..7701
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7705..7725
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7729..7749
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7750..7815
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(7896..7914)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      8072..8269
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(8870..9213)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        9261..9529
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        9549..9624
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(11744..11764)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11744..11762)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11783..11799)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11783..11799)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11796..11818)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11807..11823
                     /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(11831..11861)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11876..11897
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12014..12031)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12185..12773)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12265..12284)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12944..13804)
                     /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     13567..13586
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13805..13909)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    13946..14449
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     14490..14508
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gaattcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagatc
       61 tcggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      121 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtaccg
      181 agagtgcacc ataccacctt ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggtgatg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     1861 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     1921 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
     1981 ctcactatag ggcgaattgg gtacctgcca ctactaccac tgggagtttc gttcttctcg
     2041 atactattag ctttacttcc tgcactagca gtggttggat caacagaatc ttcataatca
     2101 tcaaaatcgt cttttgaaga ccccccgttt gatgtatggc cctgtctttt catcaaactt
     2161 tttatatagt tgactgaact gaggctaaat atgtgatcat cttcactata gacaatcttt
     2221 ctcttatttg caccaccgcc accaccagtc tttgagaaat tctcaaaacc ttttacgata
     2281 ttaccaagcg ggctctcttc gaaataatct atctcttttt gatatatcga atcctctagc
     2341 gtggttagct ttctagttag ttcttgcttc ttaagaattt gctggattag tttatttttc
     2401 aattcaacgt atttctcaga gtcatcttta gattttgatg aagatgtgcg ttcattcgct
     2461 atatccttct tggtcgtgtc ttttcgatcc tccttggctg gcactgaact cgtctttttt
     2521 ggcgttgctg ttccagacag acttatctca ttagatttgg aacttgtggg tttaacatca
     2581 tttgcatctt tagtagacat gattgtgtaa taccgtgatt gtttgttttg aaaggtctgt
     2641 catatttcta tcaatttcaa aacaaaatgt tcatcagaaa aaagccaaaa atgtctcttc
     2701 tagcttctta gtggtgtcgc aaaatacaca atgtcgctca acaatccaca ttcccggcgc
     2761 atagctcaaa tcacatgact acagctaaca attacacaaa aaaattctct tttcgatgta
     2821 gcaactatct tcaactaaaa cattttctcc ttcggcccat gattgtcctc cgggtcgaca
     2881 gcaagccgtt acaattgaga tggaaagcga cctaccttca ctcgataagg tgcttaattg
     2941 tacttcatat aaatctggcc cggatctaaa caaatgagtt ccattaagcc gtgggttctc
     3001 aattagggtt tttgtttttg atttagaaaa aagagatcaa gatttgttta caggtgatgc
     3061 ctttttttag aacttatgcg ttgcaaaagt tgactaacga tttctataag gtgatccaca
     3121 ctaattatac aaacgtacaa acagacatac ttttcctgcg ttcacctgat gttggccaga
     3181 tttctctctt cattgcatag aacataacca cactagggca acagaaaaaa aaaaaaaaag
     3241 tgcatcggga agttgtgttc cattcattat atgtctacta ctgcatatga gtagcccacc
     3301 caccaccacc atagtaagtt tttgtgtatg cgcgccgtca ggttatttca tttctgaatt
     3361 tttcaaccac cttactccct ttattgttga ttgacaattt tgctcacagt aagatctttt
     3421 agactccaat taatataaaa taagtctgat tttccaattc ctgttttttc tttttttttc
     3481 tgtttctatt tctttccttt tctccctttt ttttaattct tcattcaatc atcaattgat
     3541 aattcaggaa tattacaacc cgggatggat aaaaagtata gtattggttt agatattggt
     3601 actaactctg tgggttgggc agttatcact gacgaatata aagttccatc aaagaaattt
     3661 aaggtgttag gtaacactga cagacactca ataaaaaaga atcttatcgg tgctcttttg
     3721 ttcgactccg gtgaaactgc cgaggctaca cgtttaaaaa gaacagcaag aagaagatat
     3781 acccgtagaa aaaatagaat atgttattta caagaaatct tttctaatga aatggctaaa
     3841 gttgatgatt cctttttcca tagattggaa gagtcatttt tggttgaaga agacaaaaag
     3901 catgagagac atccaatctt tgggaatata gttgatgaag tggcttacca tgaaaaatat
     3961 cctaccattt atcatttaag aaagaaattg gtagattcaa ctgataaagc tgaccttaga
     4021 ttaatctatt tagcacttgc ccatatgatt aaatttagag gtcatttttt gattgaaggt
     4081 gatttgaacc cagataattc tgacgtggat aaattattta ttcaattagt ccaaacctac
     4141 aaccaattat ttgaggaaaa tccaattaat gctagtggtg tcgatgccaa agctatatta
     4201 tcagccagat tatcaaaatc tagacgtttg gaaaatttga ttgcccaatt gccaggagaa
     4261 aaaaagaatg gattatttgg aaacttgatc gcattatcat tgggtttgac accaaatttt
     4321 aaatctaatt ttgatttagc tgaagatgct aaattacaat tatcaaaaga cacctatgac
     4381 gacgatttgg acaatttact tgctcaaatt ggtgatcaat atgcagattt gttcttagct
     4441 gctaaaaact tatctgatgc tattttgttg tctgatattt tgagagtgaa cacagaaata
     4501 accaaagctc cattatcagc atctatgatc aaacgttatg atgaacacca tcaggatttg
     4561 actttattga aagctttggt gagacaacaa ttgccagaga agtataaaga aatctttttc
     4621 gatcaatcta aaaacgggta tgcaggttat attgatgggg gtgcctccca agaggaattt
     4681 tacaaattta taaaacctat tttagaaaag atggatggga ctgaggaact tttggtcaaa
     4741 ttgaacagag aagatttgtt acgtaaacag agaacttttg ataatggtag tatacctcac
     4801 caaattcatt tgggtgagtt gcatgcaatt ttaagaagac aagaagattt ttatccattt
     4861 ttaaaagata atagagaaaa aatcgagaaa attttaacct ttagaattcc atactatgtt
     4921 gggcctttgg ctagaggtaa ttcaagattt gcctggatga cacgtaaatc agaagaaact
     4981 attacccctt ggaattttga agaggttgtt gataaaggag catcagcaca gagttttatt
     5041 gaaagaatga ccaatttcga taaaaactta ccaaatgaaa aagttttacc aaaacattcc
     5101 ttgttatacg aatattttac tgtttacaat gaacttacaa aggttaaata tgttactgaa
     5161 ggtatgcgta agccagcctt tttatctgga gaacagaaaa aggcaatagt tgatttattg
     5221 tttaaaacaa atagaaaagt tactgttaaa caattaaaag aagattactt taagaaaatt
     5281 gaatgttttg attcagttga aatcagtggt gttgaagaca gatttaatgc tagtttagga
     5341 acttaccatg atttacttaa aattatcaaa gataaagatt tcttggataa cgaagaaaat
     5401 gaagacattt tagaagacat tgttttaacc ttaactttat tcgaagatag agagatgatt
     5461 gaagaacgtt tgaagactta tgcacatttg tttgacgata aagtgatgaa acagttgaaa
     5521 agaagacgtt atactggatg gggtagattg tctcgtaaat tgatcaatgg aattagagat
     5581 aaacaaagtg gtaaaactat cttggacttt ttgaaatctg acggatttgc taatagaaat
     5641 ttcatgcaat tgatccacga cgatagtttg acatttaaag aagacatcca aaaggcccaa
     5701 gtgagtgggc aaggtgattc attacatgaa catattgcaa atttagccgg atctcctgct
     5761 attaagaaag ggatattaca aactgttaaa gttgtggatg aattagtgaa agtaatggga
     5821 agacataaac ctgaaaacat tgtcattgag atggcaagag aaaatcaaac tacacaaaaa
     5881 ggacagaaaa atagtagaga acgtatgaaa agaatagaag agggtattaa agaattgggt
     5941 agtcaaatat tgaaagaaca cccagtggaa aatacccagt tgcaaaatga aaaattatat
     6001 ctttactacc ttcaaaatgg acgtgatatg tatgttgatc aggaattaga tataaataga
     6061 ctttcagatt atgatgtaga tcatatagtt ccacaatctt tcttgaaaga tgattccata
     6121 gacaataaag tattaactag aagtgataaa aatagaggta aaagtgataa tgtcccaagt
     6181 gaggaagtcg tcaaaaagat gaaaaattac tggcgtcaac ttttgaatgc taaattaatt
     6241 actcaaagaa aatttgataa tttgactaaa gcagaaagag gtgggctttc tgaattagat
     6301 aaagccgggt tcattaaaag acaattggtc gaaactagac aaattactaa acatgttgcc
     6361 caaattttag attcccgtat gaacactaag tatgacgaaa atgataagtt aatacgtgag
     6421 gttaaagtca ttactttaaa atcaaaactt gtctctgatt tcagaaagga tttccaattc
     6481 tataaagtta gagaaattaa taattatcat catgctcatg atgcatattt gaatgctgta
     6541 gttggaactg ctttaatcaa gaaataccct aaattagaat ctgaatttgt atatggtgat
     6601 tacaaagtct atgatgttag aaagatgatt gctaaatcag aacaagaaat tggtaaagct
     6661 acagctaaat acttctttta ctctaacatt atgaatttct ttaaaacaga aattactttg
     6721 gcaaacggtg aaattagaaa aagacctctt attgaaacaa atggtgagac tggagagata
     6781 gtttgggaca aagggcgtga tttcgctact gttagaaaag ttttatcaat gccacaagtt
     6841 aacattgtaa agaaaacaga ggttcaaact ggtggtttct caaaagaaag tattttgcct
     6901 aaaagaaata gtgataaatt gattgccaga aaaaaggatt gggatccaaa gaaatatggt
     6961 ggtttcgact caccaaccgt agcctattct gttttggttg tggcaaaggt tgaaaagggt
     7021 aaaagtaaaa agcttaaatc agtaaaagaa cttttgggta ttacaataat ggaaagaagt
     7081 tcctttgaaa agaaccctat tgattttttg gaagctaaag gttataagga agtaaagaag
     7141 gacttaataa tcaaattgcc taaatattct ttatttgaat tagaaaatgg gagaaaaaga
     7201 atgttggctt ctgctggaga attgcaaaag ggtaatgaat tagcattgcc ttccaaatat
     7261 gttaacttct tgtatttagc ttcacactat gaaaagttga aagggtcacc agaagataac
     7321 gagcaaaaac aattatttgt tgaacaacac aaacactact tagatgagat tatagaacaa
     7381 attagtgaat tcagtaaaag agtgatatta gctgatgcaa atttagataa agttttgtca
     7441 gcctataaca aacatagaga taagccaatt agagaacaag cagaaaacat tattcactta
     7501 tttaccctta ccaatttagg agcacctgct gctttcaagt attttgatac aacaattgat
     7561 cgtaaaagat atacctcaac aaaagaagtc ttagacgcca ccttaattca tcaatcaatc
     7621 actggattgt atgagacaag aattgatttg tctcaattgg gtggtgatga aggggctgat
     7681 cctaagaaga aaagaaaagt tgatccaaag aaaaagcgta aggtggatcc taagaaaaag
     7741 agaaaggttg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag
     7801 gatgacgatg acaagtgata atgactgcag agatccatcg acctgccgcc aagctaattc
     7861 cgggcgaatt tctgtcgagt catgtaatta gttatgtcac gcttacattc acgccctccc
     7921 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     7981 tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     8041 tttcttggcg gcgttagtat caaatcgaca gcagtatagc gaccagcatt cacatacgat
     8101 tgacgcatga tattactttc tgcgcactta acttcgcatc tgggcagatg atgtcgaggc
     8161 gaaaaaaaat ataaatcacg ctaacatttg attaaaatag aacaactaca atataaaaaa
     8221 actatacaaa tgacaagttc ttgaaaacaa gaatcttttt attgtcagta ctgactcgag
     8281 ttattatgga catggcatag acatatacaa agcttgttca ccatcggaag cagtaccatc
     8341 gtataaagca gtatccaaac cacacaaagt gaaacccatt cttctataag catgaatagc
     8401 tggagcatta acattggtaa cttccaacca caaatgacca gcacctcttt ctctggcgaa
     8461 ttcagtagcc aaacccatca aagctctacc aacaccatga cctctatgtt ctggagcaac
     8521 ttcaatatct tcaacagtca atcttctgtt ccaaccagaa taagaaacaa caacgaaacc
     8581 agccaaatca ccatcatcac cataagcaac gaaagttcta gaatctggat caccatcttc
     8641 accagcatcg gattcatcat cggattcatc atctgggaaa accttagtca atggtggatc
     8701 aactggaact tctctcaaag tgaaaccatc accagtagca gtaactctaa aaacagtatc
     8761 ggtagtgaaa gaaccatcca aagcttcaat agcttcagca tcacctggaa cagaagttct
     8821 gtatctataa gcagtatcat ccaaagtagt agacataatt gtaggatccg gttgtttatg
     8881 ttcggatgtg atgtgagaac tgtatcctag caagatttta aaaggaagta tatgaaagaa
     8941 gaacctcagt ggcaaatcct aaccttttat atttctctac aggggcgcgg cgtggggaca
     9001 attcaacgcg tctgtgaggg gagcgtttcc ctgctcgcag gtctgcagcg aggagccgta
     9061 atttttgctt cgcgccgtgc ggccatcaaa atgtatggat gcaaatgatt atacatgggg
     9121 atgtatgggc taaatgtacg ggcgacagtc acatcatgcc cctgagctgc gcacgtcaag
     9181 actgtcaagg agggtattct gggcctccat gtcgctggcc gggtgacccg gcggggacga
     9241 ggcaagcttg atgcggccgc tctttgaaaa gataatgtat gattttgctt tcactcatat
     9301 ttatacagaa acttgatgtt ttctttcgag tatatacaag gtgattacat gtacgtttga
     9361 agtacaactc tagattttgt agtgccctct tgggctagcg gtaaaggtgc gcattttttc
     9421 acaccctaca atgttctgtt caaaagattt tggtcaaacg ctgtagaagt gaaagttggt
     9481 gcgcatgttt cggcgttcga aacttctccg cagtgaaaga taaatgatcg gagacggaat
     9541 tccgtctcgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
     9601 gaaaaagtgg caccgagtcg gtgctttttt tctcgagttt ttttatcgag tgtttaagga
     9661 taatgataac tgaagagaag aattagtttt gccgccaccg cgggtttgcc tctgattaaa
     9721 taaaaaaaag ctggtgcttt tttttctttt ataggaacat cttgaatata tgaactaatt
     9781 aaatgataat tttttaccca tctttactct taatcactga gctgcagtca aagaaaaagg
     9841 gatacagcac ctggtgaaga gatgaacgga gactaactta gacgcgttga ttcttttcaa
     9901 ttgcacattt tattaatcga tgctaacgtc tatttacata tattctttag agatattatc
     9961 tagggcttca aataatctct ggacagcaat aaaagtctct tcaaaagtat tgtataacgg
    10021 caatggggct aatctgatta catctggtct tctttcgtca cagattatag catgatcatg
    10081 caagtacgca ttaactcgtt ccatgacgtt cttgcccttt tcatcgaaat gcggttgaaa
    10141 cataatggac aattgacatc ctctttcagc tggattcaaa ggagttaaaa ttttaaaccc
    10201 aaatttggag tttgatgtac tggattgtgg tatgtaatac ttggaattcg tcaatagatc
    10261 ctgtaaaaat tgagtcaaag caacactttt ttcacgaagt ttagatactc cacccacttt
    10321 agcatacact tccaatgacg acttcacagc aacaacatca agaacagaag gatttgactg
    10381 tctgtaagaa agagccgagt ttattggatc aaactcttct aacattttga atcgttcttg
    10441 ggagttattg ccccaccaac cagctagtct aggaacgaaa ctgctcttct tgttctctat
    10501 agtgtatttt tcatgcacaa aaattccacc tatggctcca ggtcccgagt ttaaatattt
    10561 gtaggaacac caagcagcaa aatctactcc ccaatcatgt aaatttaatg gaacattccc
    10621 aactgcatgg gcaagatccc acccaacttt aatttgttgg ctcttttcct tagcgtattt
    10681 agttatttcc tctatcttga aaaattgacc agtgtagtat tggataccag gaaaacacac
    10741 tagagccaat tcatccaggt tctcatctat agccttgatt attctttctg ttttaatata
    10801 agtttcacca ggttgaactt ccaattgaat caaatgtttc tcgtcgtatc cgaacaattt
    10861 aacaatgttc aaaaatgcat agtagtcaga aggaaatgct tgtttttcaa ataaaatttt
    10921 ggttcttttc ccctcaggtt tgtaaaaatg gatcaacaat gcattcaagt ttgctgttaa
    10981 agaacccata actgcaactt cgttttcctt tgcaccaaca atgggggcta ttaatggtaa
    11041 taagggtaaa tcgatgtcta cccacggtgt taacagtttg tcaggatgat tgaaatgaga
    11101 ctcaacccct cgttcaaccc atgcattcaa ttcatcattg atagctttct ttgtattctt
    11161 aggcatcaac ccaagagagt ttccacataa ataaatagac tcagttgatg actcatattt
    11221 attatttttg atacctaatg atccaaaagt tggtatggca aactcatttt taaaagttgg
    11281 gaactttttg tccaatttct ttgcctcggc taatgacatc tgataataaa atggggttgg
    11341 agtagttggt ggtataaccg gagagataga attgaagaaa aaaatcggaa acaacaaaaa
    11401 aagttgatac ccagtattat gtgggagata attgcgaatg gtggaaaaaa aaaagacgcc
    11461 tttgggcctc aacaacaatt ctaacagctt gagagcttta caatcgagaa actatgattc
    11521 attccgtttt aatatgtatg tacttagtaa actcgtgaat tttatttgtg gtctacttta
    11581 gtactaacat aatcattgga tagtcaataa tgatggtctt ccgagactaa tgaaattcta
    11641 taccaaagtc gatattccaa cacagaaatt gctcttgcaa caagtgcacc tgttgatatc
    11701 tactagatca gcacaagagt gttcggagct ccagcttttg ttccctttag tgagggttaa
    11761 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
    11821 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
    11881 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
    11941 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
    12001 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
    12061 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
    12121 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
    12181 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
    12241 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
    12301 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
    12361 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
    12421 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
    12481 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
    12541 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
    12601 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
    12661 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
    12721 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
    12781 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
    12841 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
    12901 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
    12961 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
    13021 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
    13081 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
    13141 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
    13201 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
    13261 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
    13321 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
    13381 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
    13441 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
    13501 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
    13561 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
    13621 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
    13681 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
    13741 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
    13801 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
    13861 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
    13921 gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa aataattata atttaaattt
    13981 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
    14041 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
    14101 tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag
    14161 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
    14221 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
    14281 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
    14341 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
    14401 ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtctaagaaa
    14461 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//