LOCUS       Exported               11106 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  All-in-one CRISPR plasmid for integrating, deleting, or replacing a 
            gene in S. cerevisiae. High-copy, URA3 plasmid contains human 
            codon-optimized Cas9 and a guide RNA targeting YPRCd15c sequence 
            AATCCGAACAACAGAGCATA in yeast chromosome 14..
ACCESSION   .
VERSION     .
KEYWORDS    p426_Cas9_gRNA-YPRCd15c
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11106)
  AUTHORS   Reider Apel A, d'Espaux L, Wehrs M, Sachs D, Li RA, Tong GJ, Garber 
            M, Nnadi O, Zhuang W, Hillson NJ, Keasling JD, Mukhopadhyay A
  TITLE     A Cas9-based toolkit to program gene expression in Saccharomyces 
            cerevisiae.
  JOURNAL   Nucleic Acids Res. 2017 Jan 9;45(1):496-508. doi: 
            10.1093/nar/gkw1023. Epub 2016 Nov 28.
  PUBMED    27899650
REFERENCE   2  (bases 1 to 11106)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11106
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     116..135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     369..386
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    503..524
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        539..569
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    577..593
                     /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)."
     primer_bind     582..604
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     601..617
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     601..617
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     636..656
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        638..656
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        883..958
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1044..1748
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1758..5861
                     /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
                     FLVEEDKKNERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIYLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5874..5894
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5910..6157
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6175..6194)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6176..6194)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6204..6221)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6204..6220)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6213..6235)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6361..6816
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6448..6467)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6658..6679
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6947..7750)
                     /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"
     promoter        complement(7751..7966)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7990..8009)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8109..8131
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8169..8187)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      8228..9570
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    8560..8607
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        9597..9701
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9702..10562
                     /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     complement(9920..9939)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10733..215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
       61 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      241 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      301 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      361 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
      421 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      481 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      541 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      601 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
      661 aaaagctgga gctagtatac tctttcttca acaattaaat actctcggta gccaagttgg
      721 tttaaggcgc aagactgtaa tttatcacta cgaaatcttg agatcgggcg ttcgactcgc
      781 ccccgggaga gatggccggc atggtcccag cctcctcgct ggcgccggct gggcaacacc
      841 ttcgggtggc gaatgggact ttaatccgaa caacagagca tagttttaga gctagaaata
      901 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
      961 tttttatttt ttgtcactat tgttatgtaa aatgccacct ctgacagtat ggaacgcaaa
     1021 cttctgtcta gtggatagtc gacatccttt tgttgtttcc gggtgtacaa tatggacttc
     1081 ctcttttctg gcaaccaaac ccatacatcg ggattcctat aataccttcg ttggtctccc
     1141 taacatgtag gtggcggagg ggagatatac aatagaacag ataccagaca agacataatg
     1201 ggctaaacaa gactacacca attacactgc ctcattgatg gtggtacata acgaactaat
     1261 actgtagccc tagacttgat agccatcatc atatcgaagt ttcactaccc tttttccatt
     1321 tgccatctat tgaagtaata ataggcgcat gcaacttctt ttcttttttt ttcttttctc
     1381 tctcccccgt tgttgtctca ccatatccgc aatgacaaaa aaatgatgga agacactaaa
     1441 ggaaaaaatt aacgacaaag acagcaccaa cagatgtcgt tgttccagag ctgatgaggg
     1501 gtatctcgaa gcacacgaaa ctttttcctt ccttcattca cgcacactac tctctaatga
     1561 gcaacggtat acggccttcc ttccagttac ttgaatttga aataaaaaaa agtttgctgt
     1621 cttgctatca agtataaata gacctgcaat tattaatctt ttgtttcctc gtcattgttc
     1681 tcgttccctt tcttccttgt ttctttttct gcacaatatt tcaagctata ccaagcatac
     1741 aatcaacttg gaaaaaaatg gacaagaagt actccattgg gctcgatatc ggcacaaaca
     1801 gcgtcggttg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc
     1861 tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc ctgttcgact
     1921 ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga tatacccgca
     1981 gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct aaggtggatg
     2041 actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa aagaacgagc
     2101 gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag tacccaacca
     2161 tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg cggttgatct
     2221 atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag ggggacctga
     2281 acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact tacaatcagc
     2341 ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc ctgagcgcta
     2401 ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga
     2461 acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac tttaaatcta
     2521 acttcgacct ggccgaagat gccaagcttc aactgagcaa agacacctac gatgatgatc
     2581 tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg gcggcaaaga
     2641 acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag atcaccaaag
     2701 ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac ttgactttgc
     2761 tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt
     2821 ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat
     2881 ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta aagcttaaca
     2941 gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc
     3001 acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag
     3061 ataacaggga aaagattgag aaaatcctca catttcggat accctactat gtaggccccc
     3121 tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag accatcactc
     3181 cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga
     3241 tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt
     3301 acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga
     3361 gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga
     3421 cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag attgaatgtt
     3481 tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc
     3541 acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca
     3601 ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac
     3661 gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc
     3721 gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga gacaagcaga
     3781 gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc
     3841 agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg
     3901 gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa
     3961 agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata
     4021 agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga
     4081 agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa
     4141 tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact
     4201 acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat cggctctccg
     4261 actacgacgt ggatcatatc gtgccccagt cttttctcaa agatgattct attgataata
     4321 aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag
     4381 ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac
     4441 ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg
     4501 gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc
     4561 tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag
     4621 ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag ttttataagg
     4681 tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca
     4741 ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga gactataaag
     4801 tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta
     4861 agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg
     4921 gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg
     4981 acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag gtgaacatcg
     5041 ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga
     5101 acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg
     5161 attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta
     5221 aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga tcaagcttcg
     5281 aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa aaagacctca
     5341 tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg
     5401 ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt
     5461 tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga
     5521 agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg
     5581 aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca
     5641 ataagcacag ggataagccc atcagggagc aggcagaaaa cattatctac ttgtttactc
     5701 tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaagc
     5761 ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc
     5821 tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct gaccccaaga
     5881 agaagaggaa ggtgtgatct cttctcgagt catgtaatta gttatgtcac gcttacgttc
     5941 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     6001 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat
     6061 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct
     6121 tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc cggtacccaa ttcgccctat
     6181 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     6241 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     6301 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     6361 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     6421 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     6481 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     6541 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     6601 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     6661 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     6721 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     6781 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     6841 gcatctgtgc ggtatttcac accgcatagg gtaataactg atataattaa attgaagctc
     6901 taatttgtga gtttagtata catgcattta cttataatac agttttttag ttttgctggc
     6961 cgcatcttct caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag
     7021 catcccttcc ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga
     7081 ccacatcatc cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca
     7141 caccgggtgt cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag
     7201 caataaagcc gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat
     7261 attctccagt agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag
     7321 gttcctttgt tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca
     7381 caccgtgtgc attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact
     7441 gcaatttgac tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt
     7501 acttggcgga taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga
     7561 tatccacatg tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta
     7621 attccttggt ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga
     7681 tattaaatag cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag
     7741 ctttcgacat gatttatctt cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga
     7801 atactgggca atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc
     7861 tgtgctcctt ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaag
     7921 gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga attgaaaagg tggtatggtg
     7981 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     8041 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     8101 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     8161 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     8221 ttagtatgat ccaatatcaa aggaaatgat agcattgaag gatgagacta atccaattga
     8281 ggagtggcag catatagaac agctaaaggg tagtgctgaa ggaagcatac gataccccgc
     8341 atggaatggg ataatatcac aggaggtact agactacctt tcatcctaca taaatagacg
     8401 catataagta cgcatttaag cataaacacg cactatgccg ttcttctcat gtatatatat
     8461 atacaggcaa cacgcagata taggtgcgac gtgaacagtg agctgtatgt gcgcagctcg
     8521 cgttgcattt tcggaagcgc tcgttttcgg aaacgctttg aagttcctat tccgaagttc
     8581 ctattctcta gaaagtatag gaacttcaga gcgcttttga aaaccaaaag cgctctgaag
     8641 acgcactttc aaaaaaccaa aaacgcaccg gactgtaacg agctactaaa atattgcgaa
     8701 taccgcttcc acaaacattg ctcaaaagta tctctttgct atatatctct gtgctatatc
     8761 cctatataac ctacccatcc acctttcgct ccttgaactt gcatctaaac tcgacctcta
     8821 cattttttat gtttatctct agtattactc tttagacaaa aaaattgtag taagaactat
     8881 tcatagagtg aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt agtatataga
     8941 gacaaaatag aagaaaccgt tcataatttt ctgaccaatg aagaatcatc aacgctatca
     9001 ctttctgttc acaaagtatg cgcaatccac atcggtatag aatataatcg gggatgcctt
     9061 tatcttgaaa aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg aagtggagtc
     9121 aggctttttt tatggaagag aaaatagaca ccaaagtagc cttcttctaa ccttaacgga
     9181 cctacagtgc aaaaagttat caagagactg cattatagag cgcacaaagg agaaaaaaag
     9241 taatctaaga tgctttgtta gaaaaatagc gctctcggga tgcatttttg tagaacaaaa
     9301 aagaagtata gattctttgt tggtaaaata gcgctctcgc gttgcatttc tgttctgtaa
     9361 aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgtttta
     9421 caaaaatgaa gcacagattc ttcgttggta aaatagcgct ttcgcgttgc atttctgttc
     9481 tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg
     9541 ttctacaaaa tgaagcacag atgcttcgtt caggtggcac ttttcgggga aatgtgcgcg
     9601 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     9661 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     9721 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     9781 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     9841 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     9901 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     9961 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
    10021 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
    10081 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
    10141 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
    10201 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
    10261 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
    10321 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
    10381 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
    10441 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
    10501 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
    10561 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
    10621 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
    10681 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
    10741 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
    10801 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
    10861 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
    10921 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
    10981 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
    11041 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
    11101 aactga
//
